Features
Primers
Cut sites
CDS
misc_feature
protein_bind
Sequence
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100
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GFP
BbsI site
ATGA overhang
TGGC overhang
BbsI site
GFP forward (19 .. 43)
GFP reverse (706 .. 731)
BbsI
(14)
BbsI
(731)
GFP
749 bp
50
100
150
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250
300
350
400
450
500
550
600
650
700
GFP
ATGA overhang
TGGC overhang
BbsI site
BbsI site
GFP forward (19 .. 43)
GFP reverse (706 .. 731)
BbsI
(14)
BbsI
(731)
GFP
749 bp
20
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60
80
100
120
140
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180
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360
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700
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740
GFP
ATGA overhang
BbsI site
TGGC overhang
BbsI site
GFP forward (19 .. 43)
GFP reverse (706 .. 731)
BbsI
(14)
BbsI
(731)
GFP
749 bp
10
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40
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GFP
BbsI site
ATGA overhang
BbsI site
TGGC overhang
GFP forward (19 .. 43)
GFP reverse (706 .. 731)
BbsI
(14)
BbsI
(731)
GFP
749 bp
Both strands
Copy
10 20 30 40 50 60
AAACACGAAGACAAATGAGTAAAGGAGAAGAACTTTTCACTGGAGTTGTCCCAATTCTTG
TTTGTGCTTCTGTTTACTCATTTCCTCTTCTTGAAAAGTGACCTCAACAGGGTTAAGAAC
60
BbsI site
Met Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu
GFP
ATGA overhang
GFP forward (19 .. 43)
BbsI
70 80 90 100 110 120
TTGAATTAGATGGTGATGTTAATGGGCACAAATTTTCTGTCAGTGGAGAGGGTGAAGGTG
AACTTAATCTACCACTACAATTACCCGTGTTTAAAAGACAGTCACCTCTCCCACTTCCAC
120
Val Glu Leu Asp Gly Asp Val Asn Gly His Lys Phe Ser Val Ser Gly Glu Gly Glu Gly
GFP
130 140 150 160 170 180
ATGCAACATACGGAAAACTTACCCTTAAATTTATTTGCACTACTGGAAAACTACCTGTTC
TACGTTGTATGCCTTTTGAATGGGAATTTAAATAAACGTGATGACCTTTTGATGGACAAG
180
Asp Ala Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile Cys Thr Thr Gly Lys Leu Pro Val
GFP
190 200 210 220 230 240
CATGGCCAACACTTGTCACTACTTTCTCTTATGGTGTTCAATGCTTTTCAAGATACCCAG
GTACCGGTTGTGAACAGTGATGAAAGAGAATACCACAAGTTACGAAAAGTTCTATGGGTC
240
Pro Trp Pro Thr Leu Val Thr Thr Phe Ser Tyr Gly Val Gln Cys Phe Ser Arg Tyr Pro
GFP
250 260 270 280 290 300
ATCATATGAAACGGCATGACTTTTTCAAGAGTGCCATGCCCGAAGGTTATGTACAGGAAA
TAGTATACTTTGCCGTACTGAAAAAGTTCTCACGGTACGGGCTTCCAATACATGTCCTTT
300
Asp His Met Lys Arg His Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu
GFP
310 320 330 340 350 360
GAACTATATTTTTCAAAGATGACGGGAACTACAAGACACGTGCTGAAGTCAAGTTTGAAG
CTTGATATAAAAAGTTTCTACTGCCCTTGATGTTCTGTGCACGACTTCAGTTCAAACTTC
360
Arg Thr Ile Phe Phe Lys Asp Asp Gly Asn Tyr Lys Thr Arg Ala Glu Val Lys Phe Glu
GFP
370 380 390 400 410 420
GTGATACCCTTGTTAATAGAATCGAGTTAAAAGGTATTGATTTTAAAGAAGATGGAAACA
CACTATGGGAACAATTATCTTAGCTCAATTTTCCATAACTAAAATTTCTTCTACCTTTGT
420
Gly Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly Ile Asp Phe Lys Glu Asp Gly Asn
GFP
430 440 450 460 470 480
TTCTTGGACACAAATTGGAATACAACTATAACTCACACAATGTATACATCATGGCAGACA
AAGAACCTGTGTTTAACCTTATGTTGATATTGAGTGTGTTACATATGTAGTACCGTCTGT
480
Ile Leu Gly His Lys Leu Glu Tyr Asn Tyr Asn Ser His Asn Val Tyr Ile Met Ala Asp
GFP
490 500 510 520 530 540
AACAAAAGAATGGAATCAAAGTTAACTTCAAAATTAGACACAACATTGAAGATGGAAGCG
TTGTTTTCTTACCTTAGTTTCAATTGAAGTTTTAATCTGTGTTGTAACTTCTACCTTCGC
540
Lys Gln Lys Asn Gly Ile Lys Val Asn Phe Lys Ile Arg His Asn Ile Glu Asp Gly Ser
GFP
550 560 570 580 590 600
TTCAACTAGCAGACCATTATCAACAAAATACTCCAATTGGCGATGGCCCTGTCCTTTTAC
AAGTTGATCGTCTGGTAATAGTTGTTTTATGAGGTTAACCGCTACCGGGACAGGAAAATG
600
Val Gln Leu Ala Asp His Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly Pro Val Leu Leu
GFP
610 620 630 640 650 660
CAGACAACCATTACCTGTCCACACAATCTGCCCTTTCGAAAGATCCCAACGAAAAGAGAG
GTCTGTTGGTAATGGACAGGTGTGTTAGACGGGAAAGCTTTCTAGGGTTGCTTTTCTCTC
660
Pro Asp Asn His Tyr Leu Ser Thr Gln Ser Ala Leu Ser Lys Asp Pro Asn Glu Lys Arg
GFP
670 680 690 700 710 720
ACCACATGGTCCTTCTTGAGTTTGTAACAGCTGCTGGGATTACACATGGCATGGATGAAC
TGGTGTACCAGGAAGAACTCAAACATTGTCGACGACCCTAATGTGTACCGTACCTACTTG
720
Asp His Met Val Leu Leu Glu Phe Val Thr Ala Ala Gly Ile Thr His Gly Met Asp Glu
GFP
GFP reverse (706 .. 731)
730 740
TATACAAATAGTGGCTTGTCTTCGTGTTT
ATATGTTTATCACCGAACAGAAGCACAAA
749
Leu Tyr Lys
Ter
GFP
TGGC overhang
BbsI site
GFP reverse (706 .. 731)
BbsI
10 20 30
AAACACGAAGACAAATGAGTAAAGGAGAAG
TTTGTGCTTCTGTTTACTCATTTCCTCTTC
30
BbsI site
Met Ser Lys Gly Glu
GFP
ATGA overhang
GFP forward (19 .. 43)
BbsI
40 50 60
AACTTTTCACTGGAGTTGTCCCAATTCTTG
TTGAAAAGTGACCTCAACAGGGTTAAGAAC
60
Glu Leu Phe Thr Gly Val Val Pro Ile Leu
GFP
GFP forward (19 .. 43)
70 80 90
TTGAATTAGATGGTGATGTTAATGGGCACA
AACTTAATCTACCACTACAATTACCCGTGT
90
Val Glu Leu Asp Gly Asp Val Asn Gly His
GFP
100 110 120
AATTTTCTGTCAGTGGAGAGGGTGAAGGTG
TTAAAAGACAGTCACCTCTCCCACTTCCAC
120
Lys Phe Ser Val Ser Gly Glu Gly Glu Gly
GFP
130 140 150
ATGCAACATACGGAAAACTTACCCTTAAAT
TACGTTGTATGCCTTTTGAATGGGAATTTA
150
Asp Ala Thr Tyr Gly Lys Leu Thr Leu Lys
GFP
160 170 180
TTATTTGCACTACTGGAAAACTACCTGTTC
AATAAACGTGATGACCTTTTGATGGACAAG
180
Phe Ile Cys Thr Thr Gly Lys Leu Pro Val
GFP
190 200 210
CATGGCCAACACTTGTCACTACTTTCTCTT
GTACCGGTTGTGAACAGTGATGAAAGAGAA
210
Pro Trp Pro Thr Leu Val Thr Thr Phe Ser
GFP
220 230 240
ATGGTGTTCAATGCTTTTCAAGATACCCAG
TACCACAAGTTACGAAAAGTTCTATGGGTC
240
Tyr Gly Val Gln Cys Phe Ser Arg Tyr Pro
GFP
250 260 270
ATCATATGAAACGGCATGACTTTTTCAAGA
TAGTATACTTTGCCGTACTGAAAAAGTTCT
270
Asp His Met Lys Arg His Asp Phe Phe Lys
GFP
280 290 300
GTGCCATGCCCGAAGGTTATGTACAGGAAA
CACGGTACGGGCTTCCAATACATGTCCTTT
300
Ser Ala Met Pro Glu Gly Tyr Val Gln Glu
GFP
310 320 330
GAACTATATTTTTCAAAGATGACGGGAACT
CTTGATATAAAAAGTTTCTACTGCCCTTGA
330
Arg Thr Ile Phe Phe Lys Asp Asp Gly Asn
GFP
340 350 360
ACAAGACACGTGCTGAAGTCAAGTTTGAAG
TGTTCTGTGCACGACTTCAGTTCAAACTTC
360
Tyr Lys Thr Arg Ala Glu Val Lys Phe Glu
GFP
370 380 390
GTGATACCCTTGTTAATAGAATCGAGTTAA
CACTATGGGAACAATTATCTTAGCTCAATT
390
Gly Asp Thr Leu Val Asn Arg Ile Glu Leu
GFP
400 410 420
AAGGTATTGATTTTAAAGAAGATGGAAACA
TTCCATAACTAAAATTTCTTCTACCTTTGT
420
Lys Gly Ile Asp Phe Lys Glu Asp Gly Asn
GFP
430 440 450
TTCTTGGACACAAATTGGAATACAACTATA
AAGAACCTGTGTTTAACCTTATGTTGATAT
450
Ile Leu Gly His Lys Leu Glu Tyr Asn Tyr
GFP
460 470 480
ACTCACACAATGTATACATCATGGCAGACA
TGAGTGTGTTACATATGTAGTACCGTCTGT
480
Asn Ser His Asn Val Tyr Ile Met Ala Asp
GFP
490 500 510
AACAAAAGAATGGAATCAAAGTTAACTTCA
TTGTTTTCTTACCTTAGTTTCAATTGAAGT
510
Lys Gln Lys Asn Gly Ile Lys Val Asn Phe
GFP
520 530 540
AAATTAGACACAACATTGAAGATGGAAGCG
TTTAATCTGTGTTGTAACTTCTACCTTCGC
540
Lys Ile Arg His Asn Ile Glu Asp Gly Ser
GFP
550 560 570
TTCAACTAGCAGACCATTATCAACAAAATA
AAGTTGATCGTCTGGTAATAGTTGTTTTAT
570
Val Gln Leu Ala Asp His Tyr Gln Gln Asn
GFP
580 590 600
CTCCAATTGGCGATGGCCCTGTCCTTTTAC
GAGGTTAACCGCTACCGGGACAGGAAAATG
600
Thr Pro Ile Gly Asp Gly Pro Val Leu Leu
GFP
610 620 630
CAGACAACCATTACCTGTCCACACAATCTG
GTCTGTTGGTAATGGACAGGTGTGTTAGAC
630
Pro Asp Asn His Tyr Leu Ser Thr Gln Ser
GFP
640 650 660
CCCTTTCGAAAGATCCCAACGAAAAGAGAG
GGGAAAGCTTTCTAGGGTTGCTTTTCTCTC
660
Ala Leu Ser Lys Asp Pro Asn Glu Lys Arg
GFP
670 680 690
ACCACATGGTCCTTCTTGAGTTTGTAACAG
TGGTGTACCAGGAAGAACTCAAACATTGTC
690
Asp His Met Val Leu Leu Glu Phe Val Thr
GFP
700 710 720
CTGCTGGGATTACACATGGCATGGATGAAC
GACGACCCTAATGTGTACCGTACCTACTTG
720
Ala Ala Gly Ile Thr His Gly Met Asp Glu
GFP
GFP reverse (706 .. 731)
730 740
TATACAAATAGTGGCTTGTCTTCGTGTTT
ATATGTTTATCACCGAACAGAAGCACAAA
749
Leu Tyr Lys
Ter
GFP
TGGC overhang
BbsI site
GFP reverse (706 .. 731)
BbsI