-
Notifications
You must be signed in to change notification settings - Fork 1
/
3294lib_pETcon-SARS2-RBD_Omicron-BA1_lib-assembled.gb
658 lines (658 loc) · 34.8 KB
/
3294lib_pETcon-SARS2-RBD_Omicron-BA1_lib-assembled.gb
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
66
67
68
69
70
71
72
73
74
75
76
77
78
79
80
81
82
83
84
85
86
87
88
89
90
91
92
93
94
95
96
97
98
99
100
101
102
103
104
105
106
107
108
109
110
111
112
113
114
115
116
117
118
119
120
121
122
123
124
125
126
127
128
129
130
131
132
133
134
135
136
137
138
139
140
141
142
143
144
145
146
147
148
149
150
151
152
153
154
155
156
157
158
159
160
161
162
163
164
165
166
167
168
169
170
171
172
173
174
175
176
177
178
179
180
181
182
183
184
185
186
187
188
189
190
191
192
193
194
195
196
197
198
199
200
201
202
203
204
205
206
207
208
209
210
211
212
213
214
215
216
217
218
219
220
221
222
223
224
225
226
227
228
229
230
231
232
233
234
235
236
237
238
239
240
241
242
243
244
245
246
247
248
249
250
251
252
253
254
255
256
257
258
259
260
261
262
263
264
265
266
267
268
269
270
271
272
273
274
275
276
277
278
279
280
281
282
283
284
285
286
287
288
289
290
291
292
293
294
295
296
297
298
299
300
301
302
303
304
305
306
307
308
309
310
311
312
313
314
315
316
317
318
319
320
321
322
323
324
325
326
327
328
329
330
331
332
333
334
335
336
337
338
339
340
341
342
343
344
345
346
347
348
349
350
351
352
353
354
355
356
357
358
359
360
361
362
363
364
365
366
367
368
369
370
371
372
373
374
375
376
377
378
379
380
381
382
383
384
385
386
387
388
389
390
391
392
393
394
395
396
397
398
399
400
401
402
403
404
405
406
407
408
409
410
411
412
413
414
415
416
417
418
419
420
421
422
423
424
425
426
427
428
429
430
431
432
433
434
435
436
437
438
439
440
441
442
443
444
445
446
447
448
449
450
451
452
453
454
455
456
457
458
459
460
461
462
463
464
465
466
467
468
469
470
471
472
473
474
475
476
477
478
479
480
481
482
483
484
485
486
487
488
489
490
491
492
493
494
495
496
497
498
499
500
501
502
503
504
505
506
507
508
509
510
511
512
513
514
515
516
517
518
519
520
521
522
523
524
525
526
527
528
529
530
531
532
533
534
535
536
537
538
539
540
541
542
543
544
545
546
547
548
549
550
551
552
553
554
555
556
557
558
559
560
561
562
563
564
565
566
567
568
569
570
571
572
573
574
575
576
577
578
579
580
581
582
583
584
585
586
587
588
589
590
591
592
593
594
595
596
597
598
599
600
601
602
603
604
605
606
607
608
609
610
611
612
613
614
615
616
617
618
619
620
621
622
623
624
625
626
627
628
629
630
631
632
633
634
635
636
637
638
639
640
641
642
643
644
645
646
647
648
649
650
651
652
653
654
655
656
657
658
LOCUS 3294_pETcon_SARS 6864 bp DNA circular 15-MAR-2022
DEFINITION .
ACCESSION urn.local...34muarj-4v
VERSION urn.local...34muarj-4v
COMMENT This is the Omicron (B.1.1.529) RBD sequence, with Twist codon
optimization. It expresses much better on yeast than the original
version I cloned, which was based on the original plasmid 2649
sequence.
COMMENT The Omicron description is here:
https://github.com/cov-lineages/pango-designation/issues/343.
COMMENT
COMMENT The RBD mutations are as follows: G339D, S371L, S373P, S375F,
K417N, N440K, G446S, S477N, T478K, E484A, Q493R, G496S, Q498R,
N501Y, Y505H.
COMMENT
COMMENT I (Allie Greaney) ordered the codon-optimized RBD Omicron sequence
from Twist and cloned into NdeI/XhoI-digested plasmid 2649. I
verified the sequence of the RBD through the cloning flanks using
Sanger sequencing. I verified expression on AWY101 yeast by flow
cytometry.
COMMENT
COMMENT ApEinfo:methylated:1
FEATURES Location/Qualifiers
primer_bind 64..80
/locus_tag="T3"
/label="T3"
/ApEinfo_label="T3"
/ApEinfo_fwdcolor="#14c0bd"
/ApEinfo_revcolor="#4ec02b"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
5'UTR complement(105..112)
/locus_tag="Partial"
/label="Partial"
/ApEinfo_label="Partial"
/ApEinfo_fwdcolor="#ffd327"
/ApEinfo_revcolor="#ffd327"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
promoter 105..707
/locus_tag="Bidirectional promoter"
/label="Bidirectional promoter"
/ApEinfo_label="Bidirectional promoter"
/ApEinfo_fwdcolor="#346ee0"
/ApEinfo_revcolor="#346ee0"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
misc_feature 112..112
/locus_tag="Geneious type: misc_marker"
/label="Geneious type: misc_marker"
/ApEinfo_label="Geneious type: misc_marker"
/ApEinfo_fwdcolor="#7eff74"
/ApEinfo_revcolor="#7eff74"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
promoter complement(112..470)
/locus_tag="promoter"
/label="promoter"
/ApEinfo_label="promoter"
/ApEinfo_fwdcolor="#346ee0"
/ApEinfo_revcolor="#346ee0"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
promoter 496..707
/locus_tag="GAL1 minimal promoter"
/label="GAL1 minimal promoter"
/ApEinfo_label="GAL1 minimal promoter"
/ApEinfo_fwdcolor="#346ee0"
/ApEinfo_revcolor="#346ee0"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
primer 648..675
/note="sequence: ctttaactaatactttcaacattttcgg"
/locus_tag="TNS_o64"
/label="TNS_o64"
/ApEinfo_label="TNS_o64"
/ApEinfo_fwdcolor="#00ffff"
/ApEinfo_revcolor="#80ff00"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
misc_feature 707..707
/locus_tag="Geneious type: misc_marker(1)"
/label="Geneious type: misc_marker(1)"
/ApEinfo_label="Geneious type: misc_marker"
/ApEinfo_fwdcolor="#7eff74"
/ApEinfo_revcolor="#7eff74"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
5'UTR 707..763
/locus_tag="Partial (all but the last 4 bases)"
/label="Partial (all but the last 4 bases)"
/ApEinfo_label="Partial (all but the last 4 bases)"
/ApEinfo_fwdcolor="#ffd327"
/ApEinfo_revcolor="#ffd327"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
primer_bind 727..750
/locus_tag="Gal1seq"
/label="Gal1seq"
/ApEinfo_label="Gal1seq"
/ApEinfo_fwdcolor="#14c0bd"
/ApEinfo_revcolor="#4ec02b"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
primer 727..750
/note="sequence: aatatacctctatactttaacgtc"
/locus_tag="TNS_o29"
/label="TNS_o29"
/ApEinfo_label="TNS_o29"
/ApEinfo_fwdcolor="#00ffff"
/ApEinfo_revcolor="#80ff00"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
misc_feature 772..777
/locus_tag="EcoRI"
/label="EcoRI"
/ApEinfo_label="EcoRI"
/ApEinfo_fwdcolor="#7eff74"
/ApEinfo_revcolor="#7eff74"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
5'UTR 779..802
/locus_tag="5'UTR"
/label="5'UTR"
/ApEinfo_label="5'UTR"
/ApEinfo_fwdcolor="#ffd327"
/ApEinfo_revcolor="#ffd327"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
CDS 803..1063
/locus_tag="CDS (Aga2p)"
/label="CDS (Aga2p)"
/ApEinfo_label="CDS (Aga2p)"
/ApEinfo_fwdcolor="#e9d024"
/ApEinfo_revcolor="#e9d024"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
misc_feature 1065..1089
/locus_tag="primer: o4 Aga2p_fwd"
/label="primer: o4 Aga2p_fwd"
/ApEinfo_label="primer: o4 Aga2p_fwd"
/ApEinfo_fwdcolor="#7eff74"
/ApEinfo_revcolor="#7eff74"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
CDS 1097..1132
/locus_tag="CDS (HA tag)"
/label="CDS (HA tag)"
/ApEinfo_label="CDS (HA tag)"
/ApEinfo_fwdcolor="#e9d024"
/ApEinfo_revcolor="#e9d024"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
primer 1098..1118
/note="sequence: catacgacgttccagactacg"
/locus_tag="oTNS_065"
/label="oTNS_065"
/ApEinfo_label="oTNS_065"
/ApEinfo_fwdcolor="#00ffff"
/ApEinfo_revcolor="#80ff00"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
primer 1120..1139
/note="sequence: tctgcaggctagtggtggag"
/locus_tag="o232"
/label="o232"
/ApEinfo_label="o232"
/ApEinfo_fwdcolor="#00ffff"
/ApEinfo_revcolor="#80ff00"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
misc_feature 1120..1189
/locus_tag="Twist 5' overlap"
/label="Twist 5' overlap"
/ApEinfo_label="Twist 5' overlap"
/ApEinfo_fwdcolor="#7eff74"
/ApEinfo_revcolor="#7eff74"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
CDS 1133..1177
/locus_tag="CDS (GGS linker)"
/label="CDS (GGS linker)"
/ApEinfo_label="CDS (GGS linker)"
/ApEinfo_fwdcolor="#e9d024"
/ApEinfo_revcolor="#e9d024"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
misc_feature 1155..1162
/locus_tag="NotI"
/label="NotI"
/ApEinfo_label="NotI"
/ApEinfo_fwdcolor="#7eff74"
/ApEinfo_revcolor="#7eff74"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
primer 1169..1189
/note="sequence: ggagggtcggctagccatatg"
/locus_tag="oTNS_005"
/label="oTNS_005"
/ApEinfo_label="oTNS_005"
/ApEinfo_fwdcolor="#00ffff"
/ApEinfo_revcolor="#80ff00"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
misc_binding 1184..1189
/locus_tag="NdeI"
/label="NdeI"
/ApEinfo_label="NdeI"
/ApEinfo_fwdcolor="#ff4809"
/ApEinfo_revcolor="#ff4809"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
CDS 1190..1792
/locus_tag="Translation 1190-1792"
/label="Translation 1190-1792"
/ApEinfo_label="Translation 1190-1792"
/ApEinfo_fwdcolor="#e9d024"
/ApEinfo_revcolor="#e9d024"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
misc_feature 1214..1216
/locus_tag="G339D"
/label="G339D"
/ApEinfo_label="G339D"
/ApEinfo_fwdcolor="#7eff74"
/ApEinfo_revcolor="#7eff74"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
misc_feature 1310..1312
/locus_tag="S371L"
/label="S371L"
/ApEinfo_label="S371L"
/ApEinfo_fwdcolor="#7eff74"
/ApEinfo_revcolor="#7eff74"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
misc_feature 1316..1318
/locus_tag="S373P"
/label="S373P"
/ApEinfo_label="S373P"
/ApEinfo_fwdcolor="#7eff74"
/ApEinfo_revcolor="#7eff74"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
misc_feature 1322..1324
/locus_tag="S375F"
/label="S375F"
/ApEinfo_label="S375F"
/ApEinfo_fwdcolor="#7eff74"
/ApEinfo_revcolor="#7eff74"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
misc_feature 1448..1450
/locus_tag="K417N"
/label="K417N"
/ApEinfo_label="K417N"
/ApEinfo_fwdcolor="#7eff74"
/ApEinfo_revcolor="#7eff74"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
misc_feature 1517..1519
/locus_tag="N440K"
/label="N440K"
/ApEinfo_label="N440K"
/ApEinfo_fwdcolor="#7eff74"
/ApEinfo_revcolor="#7eff74"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
misc_feature 1535..1537
/locus_tag="G446S"
/label="G446S"
/ApEinfo_label="G446S"
/ApEinfo_fwdcolor="#7eff74"
/ApEinfo_revcolor="#7eff74"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
misc_feature 1628..1630
/locus_tag="S477N"
/label="S477N"
/ApEinfo_label="S477N"
/ApEinfo_fwdcolor="#7eff74"
/ApEinfo_revcolor="#7eff74"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
misc_feature 1631..1633
/locus_tag="T478K"
/label="T478K"
/ApEinfo_label="T478K"
/ApEinfo_fwdcolor="#7eff74"
/ApEinfo_revcolor="#7eff74"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
misc_feature 1649..1651
/locus_tag="E484A"
/label="E484A"
/ApEinfo_label="E484A"
/ApEinfo_fwdcolor="#7eff74"
/ApEinfo_revcolor="#7eff74"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
misc_feature 1676..1678
/locus_tag="Q493R"
/label="Q493R"
/ApEinfo_label="Q493R"
/ApEinfo_fwdcolor="#7eff74"
/ApEinfo_revcolor="#7eff74"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
misc_feature 1685..1687
/locus_tag="G496S"
/label="G496S"
/ApEinfo_label="G496S"
/ApEinfo_fwdcolor="#7eff74"
/ApEinfo_revcolor="#7eff74"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
misc_feature 1691..1693
/locus_tag="Q498R"
/label="Q498R"
/ApEinfo_label="Q498R"
/ApEinfo_fwdcolor="#7eff74"
/ApEinfo_revcolor="#7eff74"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
misc_feature 2156..2213
/locus_tag="IlluminaP5"
/label="IlluminaP5"
/ApEinfo_label="IlluminaP5"
/ApEinfo_fwdcolor="#00f900"
/ApEinfo_revcolor="green"
/ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
width 5 offset 0"
misc_feature 1700..1702
/locus_tag="N501Y"
/label="N501Y"
/ApEinfo_label="N501Y"
/ApEinfo_fwdcolor="#7eff74"
/ApEinfo_revcolor="#7eff74"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
misc_feature 2214..2229
/locus_tag="N16"
/label="N16"
/ApEinfo_label="N16"
/ApEinfo_fwdcolor="#ff9200"
/ApEinfo_revcolor="green"
/ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
width 5 offset 0"
misc_feature 1712..1714
/locus_tag="Y505H"
/label="Y505H"
/ApEinfo_label="Y505H"
/ApEinfo_fwdcolor="#7eff74"
/ApEinfo_revcolor="#7eff74"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
misc_feature 2230..2237
/locus_tag="NotI(1)"
/label="NotI(1)"
/ApEinfo_label="NotI"
/ApEinfo_fwdcolor="#ff2600"
/ApEinfo_revcolor="green"
/ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
width 5 offset 0"
misc_binding 1793..1798
/locus_tag="XhoI"
/label="XhoI"
/ApEinfo_label="XhoI"
/ApEinfo_fwdcolor="#ff4809"
/ApEinfo_revcolor="#ff4809"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
primer complement(1793..1812)
/note="sequence: tcggaaccgcccccctcgag"
/locus_tag="TNS_o6"
/label="TNS_o6"
/ApEinfo_label="TNS_o6"
/ApEinfo_fwdcolor="#00ffff"
/ApEinfo_revcolor="#80ff00"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
CDS 1799..1810
/locus_tag="CDS"
/label="CDS"
/ApEinfo_label="CDS"
/ApEinfo_fwdcolor="#e9d024"
/ApEinfo_revcolor="#e9d024"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
CDS 1811..1840
/locus_tag="CDS (Myc tag)"
/label="CDS (Myc tag)"
/ApEinfo_label="CDS (Myc tag)"
/ApEinfo_fwdcolor="#e9d024"
/ApEinfo_revcolor="#e9d024"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
3'UTR 1851..2155
/locus_tag="yeast mating factor alpha 3' UTR"
/label="yeast mating factor alpha 3' UTR"
/ApEinfo_label="yeast mating factor alpha 3' UTR"
/ApEinfo_fwdcolor="#f76ca1"
/ApEinfo_revcolor="#008040"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
primer 1881..1910
/note="sequence: cgactttgttcccactgtacttttagctcg"
/locus_tag="AG_211"
/label="AG_211"
/ApEinfo_label="AG_211"
/ApEinfo_fwdcolor="#00ffff"
/ApEinfo_revcolor="#80ff00"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
primer complement(1881..1910)
/note="sequence: cgagctaaaagtacagtgggaacaaagtcg"
/locus_tag="AG_212"
/label="AG_212"
/ApEinfo_label="AG_212"
/ApEinfo_fwdcolor="#00ffff"
/ApEinfo_revcolor="#80ff00"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
misc_feature 2238..2243
/locus_tag="SacI"
/label="SacI"
/ApEinfo_label="SacI"
/ApEinfo_fwdcolor="#7eff74"
/ApEinfo_revcolor="#7eff74"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
primer_bind complement(2251..2270)
/locus_tag="T7"
/label="T7"
/ApEinfo_label="T7"
/ApEinfo_fwdcolor="#14c0bd"
/ApEinfo_revcolor="#4ec02b"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
misc_feature 2290..2323
/locus_tag="Illumina P7 handle"
/label="Illumina P7 handle"
/ApEinfo_label="Illumina P7 handle"
/ApEinfo_fwdcolor="#7eff74"
/ApEinfo_revcolor="#7eff74"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
rep_origin 2468..2923
/locus_tag="rep_origin"
/label="rep_origin"
/ApEinfo_label="rep_origin"
/ApEinfo_fwdcolor="#999999"
/ApEinfo_revcolor="#999999"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
misc_feature 2593..2598
/locus_tag="NaeI"
/label="NaeI"
/ApEinfo_label="NaeI"
/ApEinfo_fwdcolor="#7eff74"
/ApEinfo_revcolor="#7eff74"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
3'UTR complement(2980..3025)
/locus_tag="3'UTR"
/label="3'UTR"
/ApEinfo_label="3'UTR"
/ApEinfo_fwdcolor="#f76ca1"
/ApEinfo_revcolor="#008040"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
CDS complement(3026..3700)
/locus_tag="Tyrosinase-related protein 1 (YeastSelectable
marker)"
/label="Tyrosinase-related protein 1 (YeastSelectable
marker)"
/ApEinfo_label="Tyrosinase-related protein 1
(YeastSelectable marker)"
/ApEinfo_fwdcolor="#e9d024"
/ApEinfo_revcolor="#e9d024"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
5'UTR complement(3701..3804)
/locus_tag="5'UTR(1)"
/label="5'UTR(1)"
/ApEinfo_label="5'UTR"
/ApEinfo_fwdcolor="#ffd327"
/ApEinfo_revcolor="#ffd327"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
rep_origin complement(4233..4746)
/locus_tag="rep_origin(1)"
/label="rep_origin(1)"
/ApEinfo_label="rep_origin"
/ApEinfo_fwdcolor="#999999"
/ApEinfo_revcolor="#999999"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
rep_origin complement(4237..4613)
/locus_tag="rep_origin(2)"
/label="rep_origin(2)"
/ApEinfo_label="rep_origin"
/ApEinfo_fwdcolor="#999999"
/ApEinfo_revcolor="#999999"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
centromere complement(4626..4651)
/locus_tag="CDEI fragment of chromosome VI centromerefrom
S.cerevisiae"
/label="CDEI fragment of chromosome VI centromerefrom
S.cerevisiae"
/ApEinfo_label="CDEI fragment of chromosome VI
centromerefrom S.cerevisiae"
/ApEinfo_fwdcolor="pink"
/ApEinfo_revcolor="pink"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
promoter 4815..4843
/locus_tag="promoter(1)"
/label="promoter(1)"
/ApEinfo_label="promoter"
/ApEinfo_fwdcolor="#346ee0"
/ApEinfo_revcolor="#346ee0"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
CDS 4885..5745
/locus_tag="CDS(1)"
/label="CDS(1)"
/ApEinfo_label="CDS"
/ApEinfo_fwdcolor="#e9d024"
/ApEinfo_revcolor="#e9d024"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
rep_origin 5893..6678
/locus_tag="rep_origin(3)"
/label="rep_origin(3)"
/ApEinfo_label="rep_origin"
/ApEinfo_fwdcolor="#999999"
/ApEinfo_revcolor="#999999"
/ApEinfo_graphicformat="arrow_data {{0 0.5 0 1 2 0 0 -1 0
-0.5} {} 0} width 5 offset 0"
ORIGIN
1 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc
61 gaaattaacc ctcactaaag ggaacaaaag ctggtaccaa ttccttgaat tttcaaaaat
121 tcttactttt tttttggatg gacgcaaaga agtttaataa tcatattaca tggcattacc
181 accatataca tatccatatc taatcttact tatatgttgt ggaaatgtaa agagccccat
241 tatcttagcc taaaaaaacc ttctctttgg aactttcagt aatacgctta actgctcatt
301 gctatattga agtacggatt agaagccgcc gagcgggtga cagccctccg aaggaagact
361 ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg tgcctcgcgc
421 cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat gaagaggaaa
481 aattggcagt aacctggccc cacaaacctt caaatgaacg aatcaaatta acaaccatag
541 gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag cgaagcgatg
601 atttttgatc tattaacaga tatataaatg caaaaactgc ataaccactt taactaatac
661 tttcaacatt ttcggtttgt attacttctt attcaaatgt aataaaagta tcaacaaaaa
721 attgttaata tacctctata ctttaacgtc aaggagaaaa aaccccggat cgaattccct
781 acttcataca ttttcaatta agatgcagtt acttcgctgt ttttcaatat tttctgttat
841 tgcttcagtt ttagcacagg aactgacaac tatatgcgag caaatcccct caccaacttt
901 agaatcgacg ccgtactctt tgtcaacgac tactattttg gccaacggga aggcaatgca
961 aggagttttt gaatattaca aatcagtaac gtttgtcagt aattgcggtt ctcacccctc
1021 aacaactagc aaaggcagcc ccataaacac acagtatgtt tttaaggaca atagctcgac
1081 gattgaaggt agatacccat acgacgttcc agactacgct ctgcaggcta gtggtggagg
1141 aggctctggt ggaggcggCC gcggaggcgg agggtcggct agccatatgA ACATAACAAA
1201 CTTATGCCCC TTTGACGAAG TATTTAATGC TACTAGATTC GCATCGGTTT ATGCCTGGAA
1261 TAGAAAGAGG ATCAGTAACT GCGTTGCTGA TTATTCTGTT TTGTATAACC TGGCCCCTTT
1321 CTTCACATTT AAGTGCTACG GGGTCTCGCC TACCAAATTA AACGATTTAT GCTTCACCAA
1381 TGTGTACGCC GATTCTTTTG TGATCAGGGG TGACGAAGTT AGACAGATCG CTCCAGGGCA
1441 AACTGGTAAT ATTGCCGATT ACAACTACAA GCTTCCAGAC GACTTCACTG GTTGCGTAAT
1501 AGCATGGAAC TCAAACAAGT TAGACTCAAA GGTCTCAGGA AATTATAACT ACCTGTACCG
1561 TCTTTTCAGG AAATCAAATT TGAAGCCGTT CGAAAGGGAC ATCTCCACGG AGATATATCA
1621 GGCCGGTAAC AAGCCCTGCA ATGGCGTCGC CGGCTTCAAC TGTTACTTCC CCCTAAGGTC
1681 ATACTCTTTC AGGCCTACAT ACGGAGTTGG CCATCAGCCA TACAGAGTTG TGGTTTTATC
1741 TTTCGAGTTG TTGCACGCCC CTGCTACGGT TTGTGGTCCT AAGAAGTCCA CTctcgaggg
1801 gggcggttcc gaacaaaagc ttatttctga agaggacttg taatagagat ctgataacaa
1861 cagtgtagat gtaacaaaat cgactttgtt cccactgtac ttttagctcg tacaaaatac
1921 aatatacttt tcatttctcc gtaaacaaca tgttttccca tgtaatatcc ttttctattt
1981 ttcgttccgt taccaacttt acacatactt tatatagcta ttcacttcta tacactaaaa
2041 aactaagaca attttaattt tgctgcctgc catatttcaa tttgttataa attcctataa
2101 tttatcctat tagtagctaa aaaaagatga atgtgaatcg aatcctaaga gaattAATGA
2161 TACGGCGACC ACCGAGATCT ACACTCTTTC CCTACACGAC GCTCTTCCGA TCTNNNNNNN
2221 NNNNNNNNNG CGGCCGCgag ctccaattcg ccctatagtg agtcgtatta caattcactg
2281 gccgtcgttA GATCGGAAGA GCACACGTCT GAACTCCAGT CACttacaac gtcgtgactg
2341 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
2401 gcgtaatagc gaagaggccc gcaccgatcg cctttcccaa cagttgcgca gcctgaatgg
2461 cgaatggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
2521 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
2581 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
2641 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
2701 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
2761 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
2821 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
2881 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc
2941 tccttacgca tctgtgcggt atttcacacc gcatagatcg gcaagtgcac aaacaatact
3001 taaataaata ctactcagta ataacctatt tcttagcatt tttgacgaaa tttgctattt
3061 tgttagagtc ttttacacca tttgtctcca cacctccgct tacatcaaca ccaataacgc
3121 catttaatct aagcgcatca ccaacatttt ctggcgtcag tccaccagct aacataaaat
3181 gtaagctttc ggggctctct tgccttccaa cccagtcaga aatcgagttc caatccaaaa
3241 gttcacctgt cccacctgct tctgaatcaa acaagggaat aaacgaatga ggtttctgtg
3301 aagctgcact gagtagtatg ttgcagtctt ttggaaatac gagtctttta ataactggca
3361 aaccgaggaa ctcttggtat tcttgccacg actcatctcc atgcagttgg acgatatcaa
3421 tgccgtaatc attgaccaga gccaaaacat cctccttagg ttgattacga aacacgccaa
3481 ccaagtattt cggagtgcct gaactatttt tatatgcttt tacaagactt gaaattttcc
3541 ttgcaataac cgggtcaatt gttctctttc tattgggcac acatataata cccagcaagt
3601 cagcatcgga atctagagca cattctgcgg cctctgtgct ctgcaagccg caaactttca
3661 ccaatggacc agaactacct gtgaaattaa taacagacat actccaagct gcctttgtgt
3721 gcttaatcac gtatactcac gtgctcaata gtcaccaatg ccctccctct tggccctctc
3781 cttttctttt ttcgaccgaa ttaattctta atcggcaaaa aaagaaaagc tccggatcaa
3841 gattgtacgt aaggtgacaa gctatttttc aataaagaat atcttccact actgccatct
3901 ggcgtcataa ctgcaaagta cacatatatt acgatgctgt tctattaaat gcttcctata
3961 ttatatatat agtaatgtcg tgatctatgg tgcactctca gtacaatctg ctctgatgcc
4021 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
4081 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
4141 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
4201 ttataggtta atgtcatgat aataatggtt tcttagacgg atcgcttgcc tgtaacttac
4261 acgcgcctcg tatcttttaa tgatggaata atttgggaat ttactctgtg tttatttatt
4321 tttatgtttt gtatttggat tttagaaagt aaataaagaa ggtagaagag ttacggaatg
4381 aagaaaaaaa aataaacaaa ggtttaaaaa atttcaacaa aaagcgtact ttacatatat
4441 atttattaga caagaaaagc agattaaata gatatacatt cgattaacga taagtaaaat
4501 gtaaaatcac aggattttcg tgtgtggtct tctacacaga caagatgaaa caattcggca
4561 ttaatacctg agagcaggaa gagcaagata aaaggtagta tttgttggcg atccccctag
4621 agtcttttac atcttcggaa aacaaaaact attttttctt taatttcttt ttttactttc
4681 tatttttaat ttatatattt atattaaaaa atttaaatta taattatttt tatagcacgt
4741 gatgaaaagg acccaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
4801 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
4861 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
4921 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
4981 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
5041 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
5101 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
5161 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
5221 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
5281 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
5341 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
5401 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
5461 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
5521 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
5581 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
5641 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
5701 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
5761 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
5821 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
5881 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
5941 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
6001 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
6061 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
6121 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
6181 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
6241 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
6301 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
6361 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
6421 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
6481 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
6541 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
6601 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
6661 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
6721 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
6781 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
6841 cttccggctc gtatgttgtg tgga
//