Features
Primers
CDS
misc_recomb
Circle
Line
Sequence
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200
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600
800
1000
KanR
GFP
attL1
attL2
attB-GFP forward (161 .. 186)
attB-GFP reverse (852 .. 877)
pDONR-demo-attB-GFP
1037 bp
200
400
600
800
1000
KanR
attL1
GFP
attL2
attB-GFP forward (161 .. 186)
attB-GFP reverse (852 .. 877)
pDONR-demo-attB-GFP
1037 bp
100
200
300
400
500
600
700
800
900
1000
KanR
attL1
GFP
attL2
attB-GFP forward (161 .. 186)
attB-GFP reverse (852 .. 877)
pDONR-demo-attB-GFP
1037 bp
50
100
150
200
250
300
350
400
450
500
550
600
650
700
750
800
850
900
950
1000
KanR
attL1
GFP
attL2
attB-GFP forward (161 .. 186)
attB-GFP reverse (852 .. 877)
pDONR-demo-attB-GFP
1037 bp
20
40
60
80
100
120
140
160
180
200
220
240
260
280
300
320
340
360
380
400
420
440
460
480
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520
540
560
580
600
620
640
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680
700
720
740
760
780
800
820
840
860
880
900
920
940
960
980
1000
1020
KanR
attL1
GFP
attL2
attB-GFP forward (161 .. 186)
attB-GFP reverse (852 .. 877)
pDONR-demo-attB-GFP
1037 bp
Both strands
Copy
10 20 30 40 50 60
TTGACCGCTTAAGGCTAACGTCAGTTGCATGCCTTAAGGCTTGACCAATGCGTTCAGGCT
AACTGGCGAATTCCGATTGCAGTCAACGTACGGAATTCCGAACTGGTTACGCAAGTCCGA
60
Leu Thr Ala Gly Arg Gln Leu His Ala Leu Arg Leu Asp Gln Cys Val Gln Ala
Ter Ter
KanR
70 80 90 100 110 120
CAAATAATGATTTTATTTTGACTGATAGTGACCTGTTCGTTGCAACAAATTGATAAGCAA
GTTTATTACTAAAATAAAACTGACTATCACTGGACAAGCAACGTTGTTTAACTATTCGTT
120
130 140 150 160 170 180
TGCTTTTTTATAATGCCAACTTTGTACAAAAAAGCAGGCTATGAGTAAAGGAGAAGAACT
ACGAAAAAATATTACGGTTGAAACATGTTTTTTCGTCCGATACTCATTTCCTCTTCTTGA
180
attL1
Met Ser Lys Gly Glu Glu Leu
GFP
attB-GFP forward (161 .. 186)
190 200 210 220 230 240
TTTCACTGGAGTTGTCCCAATTCTTGTTGAATTAGATGGTGATGTTAATGGGCACAAATT
AAAGTGACCTCAACAGGGTTAAGAACAACTTAATCTACCACTACAATTACCCGTGTTTAA
240
Phe Thr Gly Val Val Pro Ile Leu Val Glu Leu Asp Gly Asp Val Asn Gly His Lys Phe
GFP
attB-GFP forward (161 .. 186)
250 260 270 280 290 300
TTCTGTCAGTGGAGAGGGTGAAGGTGATGCAACATACGGAAAACTTACCCTTAAATTTAT
AAGACAGTCACCTCTCCCACTTCCACTACGTTGTATGCCTTTTGAATGGGAATTTAAATA
300
Ser Val Ser Gly Glu Gly Glu Gly Asp Ala Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile
GFP
310 320 330 340 350 360
TTGCACTACTGGAAAACTACCTGTTCCATGGCCAACACTTGTCACTACTTTCTCTTATGG
AACGTGATGACCTTTTGATGGACAAGGTACCGGTTGTGAACAGTGATGAAAGAGAATACC
360
Cys Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr Phe Ser Tyr Gly
GFP
370 380 390 400 410 420
TGTTCAATGCTTTTCAAGATACCCAGATCATATGAAACGGCATGACTTTTTCAAGAGTGC
ACAAGTTACGAAAAGTTCTATGGGTCTAGTATACTTTGCCGTACTGAAAAAGTTCTCACG
420
Val Gln Cys Phe Ser Arg Tyr Pro Asp His Met Lys Arg His Asp Phe Phe Lys Ser Ala
GFP
430 440 450 460 470 480
CATGCCCGAAGGTTATGTACAGGAAAGAACTATATTTTTCAAAGATGACGGGAACTACAA
GTACGGGCTTCCAATACATGTCCTTTCTTGATATAAAAAGTTTCTACTGCCCTTGATGTT
480
Met Pro Glu Gly Tyr Val Gln Glu Arg Thr Ile Phe Phe Lys Asp Asp Gly Asn Tyr Lys
GFP
490 500 510 520 530 540
GACACGTGCTGAAGTCAAGTTTGAAGGTGATACCCTTGTTAATAGAATCGAGTTAAAAGG
CTGTGCACGACTTCAGTTCAAACTTCCACTATGGGAACAATTATCTTAGCTCAATTTTCC
540
Thr Arg Ala Glu Val Lys Phe Glu Gly Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly
GFP
550 560 570 580 590 600
TATTGATTTTAAAGAAGATGGAAACATTCTTGGACACAAATTGGAATACAACTATAACTC
ATAACTAAAATTTCTTCTACCTTTGTAAGAACCTGTGTTTAACCTTATGTTGATATTGAG
600
Ile Asp Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr Asn Tyr Asn Ser
GFP
610 620 630 640 650 660
ACACAATGTATACATCATGGCAGACAAACAAAAGAATGGAATCAAAGTTAACTTCAAAAT
TGTGTTACATATGTAGTACCGTCTGTTTGTTTTCTTACCTTAGTTTCAATTGAAGTTTTA
660
His Asn Val Tyr Ile Met Ala Asp Lys Gln Lys Asn Gly Ile Lys Val Asn Phe Lys Ile
GFP
670 680 690 700 710 720
TAGACACAACATTGAAGATGGAAGCGTTCAACTAGCAGACCATTATCAACAAAATACTCC
ATCTGTGTTGTAACTTCTACCTTCGCAAGTTGATCGTCTGGTAATAGTTGTTTTATGAGG
720
Arg His Asn Ile Glu Asp Gly Ser Val Gln Leu Ala Asp His Tyr Gln Gln Asn Thr Pro
GFP
730 740 750 760 770 780
AATTGGCGATGGCCCTGTCCTTTTACCAGACAACCATTACCTGTCCACACAATCTGCCCT
TTAACCGCTACCGGGACAGGAAAATGGTCTGTTGGTAATGGACAGGTGTGTTAGACGGGA
780
Ile Gly Asp Gly Pro Val Leu Leu Pro Asp Asn His Tyr Leu Ser Thr Gln Ser Ala Leu
GFP
790 800 810 820 830 840
TTCGAAAGATCCCAACGAAAAGAGAGACCACATGGTCCTTCTTGAGTTTGTAACAGCTGC
AAGCTTTCTAGGGTTGCTTTTCTCTCTGGTGTACCAGGAAGAACTCAAACATTGTCGACG
840
Ser Lys Asp Pro Asn Glu Lys Arg Asp His Met Val Leu Leu Glu Phe Val Thr Ala Ala
GFP
850 860 870 880 890 900
TGGGATTACACATGGCATGGATGAACTATACAAATAGACCCAGCTTTCTTGTACAAAGTT
ACCCTAATGTGTACCGTACCTACTTGATATGTTTATCTGGGTCGAAAGAACATGTTTCAA
900
Gly Ile Thr His Gly Met Asp Glu Leu Tyr Lys
Ter
GFP
attL2
attB-GFP reverse (852 .. 877)
910 920 930 940 950 960
GGCATTATAAGAAAGCATTGCTTATCAATTTGTTGCAACGAACAGGTCACTATCAGTCAA
CCGTAATATTCTTTCGTAACGAATAGTTAAACAACGTTGCTTGTCCAGTGATAGTCAGTT
960
attL2
970 980 990 1000 1010 1020
AATAAAATCATTATTTGTTGACCGCTTAAGGCTAACGTCAGTTGCATGCCTTAAGGCTTG
TTATTTTAGTAATAAACAACTGGCGAATTCCGATTGCAGTCAACGTACGGAATTCCGAAC
1020
1030
ACCAATGCGTTCAGGCT
TGGTTACGCAAGTCCGA
1037
10 20 30
TTGACCGCTTAAGGCTAACGTCAGTTGCAT
AACTGGCGAATTCCGATTGCAGTCAACGTA
30
Leu Thr Ala Gly Arg Gln Leu His
Ter Ter
KanR
40 50 60
GCCTTAAGGCTTGACCAATGCGTTCAGGCT
CGGAATTCCGAACTGGTTACGCAAGTCCGA
60
Ala Leu Arg Leu Asp Gln Cys Val Gln Ala
KanR
70 80 90
CAAATAATGATTTTATTTTGACTGATAGTG
GTTTATTACTAAAATAAAACTGACTATCAC
90
100 110 120
ACCTGTTCGTTGCAACAAATTGATAAGCAA
TGGACAAGCAACGTTGTTTAACTATTCGTT
120
130 140 150
TGCTTTTTTATAATGCCAACTTTGTACAAA
ACGAAAAAATATTACGGTTGAAACATGTTT
150
attL1
160 170 180
AAAGCAGGCTATGAGTAAAGGAGAAGAACT
TTTCGTCCGATACTCATTTCCTCTTCTTGA
180
attL1
Met Ser Lys Gly Glu Glu Leu
GFP
attB-GFP forward (161 .. 186)
190 200 210
TTTCACTGGAGTTGTCCCAATTCTTGTTGA
AAAGTGACCTCAACAGGGTTAAGAACAACT
210
Phe Thr Gly Val Val Pro Ile Leu Val Glu
GFP
attB-GFP forward (161 .. 186)
220 230 240
ATTAGATGGTGATGTTAATGGGCACAAATT
TAATCTACCACTACAATTACCCGTGTTTAA
240
Leu Asp Gly Asp Val Asn Gly His Lys Phe
GFP
250 260 270
TTCTGTCAGTGGAGAGGGTGAAGGTGATGC
AAGACAGTCACCTCTCCCACTTCCACTACG
270
Ser Val Ser Gly Glu Gly Glu Gly Asp Ala
GFP
280 290 300
AACATACGGAAAACTTACCCTTAAATTTAT
TTGTATGCCTTTTGAATGGGAATTTAAATA
300
Thr Tyr Gly Lys Leu Thr Leu Lys Phe Ile
GFP
310 320 330
TTGCACTACTGGAAAACTACCTGTTCCATG
AACGTGATGACCTTTTGATGGACAAGGTAC
330
Cys Thr Thr Gly Lys Leu Pro Val Pro Trp
GFP
340 350 360
GCCAACACTTGTCACTACTTTCTCTTATGG
CGGTTGTGAACAGTGATGAAAGAGAATACC
360
Pro Thr Leu Val Thr Thr Phe Ser Tyr Gly
GFP
370 380 390
TGTTCAATGCTTTTCAAGATACCCAGATCA
ACAAGTTACGAAAAGTTCTATGGGTCTAGT
390
Val Gln Cys Phe Ser Arg Tyr Pro Asp His
GFP
400 410 420
TATGAAACGGCATGACTTTTTCAAGAGTGC
ATACTTTGCCGTACTGAAAAAGTTCTCACG
420
Met Lys Arg His Asp Phe Phe Lys Ser Ala
GFP
430 440 450
CATGCCCGAAGGTTATGTACAGGAAAGAAC
GTACGGGCTTCCAATACATGTCCTTTCTTG
450
Met Pro Glu Gly Tyr Val Gln Glu Arg Thr
GFP
460 470 480
TATATTTTTCAAAGATGACGGGAACTACAA
ATATAAAAAGTTTCTACTGCCCTTGATGTT
480
Ile Phe Phe Lys Asp Asp Gly Asn Tyr Lys
GFP
490 500 510
GACACGTGCTGAAGTCAAGTTTGAAGGTGA
CTGTGCACGACTTCAGTTCAAACTTCCACT
510
Thr Arg Ala Glu Val Lys Phe Glu Gly Asp
GFP
520 530 540
TACCCTTGTTAATAGAATCGAGTTAAAAGG
ATGGGAACAATTATCTTAGCTCAATTTTCC
540
Thr Leu Val Asn Arg Ile Glu Leu Lys Gly
GFP
550 560 570
TATTGATTTTAAAGAAGATGGAAACATTCT
ATAACTAAAATTTCTTCTACCTTTGTAAGA
570
Ile Asp Phe Lys Glu Asp Gly Asn Ile Leu
GFP
580 590 600
TGGACACAAATTGGAATACAACTATAACTC
ACCTGTGTTTAACCTTATGTTGATATTGAG
600
Gly His Lys Leu Glu Tyr Asn Tyr Asn Ser
GFP
610 620 630
ACACAATGTATACATCATGGCAGACAAACA
TGTGTTACATATGTAGTACCGTCTGTTTGT
630
His Asn Val Tyr Ile Met Ala Asp Lys Gln
GFP
640 650 660
AAAGAATGGAATCAAAGTTAACTTCAAAAT
TTTCTTACCTTAGTTTCAATTGAAGTTTTA
660
Lys Asn Gly Ile Lys Val Asn Phe Lys Ile
GFP
670 680 690
TAGACACAACATTGAAGATGGAAGCGTTCA
ATCTGTGTTGTAACTTCTACCTTCGCAAGT
690
Arg His Asn Ile Glu Asp Gly Ser Val Gln
GFP
700 710 720
ACTAGCAGACCATTATCAACAAAATACTCC
TGATCGTCTGGTAATAGTTGTTTTATGAGG
720
Leu Ala Asp His Tyr Gln Gln Asn Thr Pro
GFP
730 740 750
AATTGGCGATGGCCCTGTCCTTTTACCAGA
TTAACCGCTACCGGGACAGGAAAATGGTCT
750
Ile Gly Asp Gly Pro Val Leu Leu Pro Asp
GFP
760 770 780
CAACCATTACCTGTCCACACAATCTGCCCT
GTTGGTAATGGACAGGTGTGTTAGACGGGA
780
Asn His Tyr Leu Ser Thr Gln Ser Ala Leu
GFP
790 800 810
TTCGAAAGATCCCAACGAAAAGAGAGACCA
AAGCTTTCTAGGGTTGCTTTTCTCTCTGGT
810
Ser Lys Asp Pro Asn Glu Lys Arg Asp His
GFP
820 830 840
CATGGTCCTTCTTGAGTTTGTAACAGCTGC
GTACCAGGAAGAACTCAAACATTGTCGACG
840
Met Val Leu Leu Glu Phe Val Thr Ala Ala
GFP
850 860 870
TGGGATTACACATGGCATGGATGAACTATA
ACCCTAATGTGTACCGTACCTACTTGATAT
870
Gly Ile Thr His Gly Met Asp Glu Leu Tyr
GFP
attB-GFP reverse (852 .. 877)
880 890 900
CAAATAGACCCAGCTTTCTTGTACAAAGTT
GTTTATCTGGGTCGAAAGAACATGTTTCAA
900
Lys
Ter
GFP
attL2
attB-GFP reverse (852 .. 877)
910 920 930
GGCATTATAAGAAAGCATTGCTTATCAATT
CCGTAATATTCTTTCGTAACGAATAGTTAA
930
attL2
940 950 960
TGTTGCAACGAACAGGTCACTATCAGTCAA
ACAACGTTGCTTGTCCAGTGATAGTCAGTT
960
970 980 990
AATAAAATCATTATTTGTTGACCGCTTAAG
TTATTTTAGTAATAAACAACTGGCGAATTC
990
1000 1010 1020
GCTAACGTCAGTTGCATGCCTTAAGGCTTG
CGATTGCAGTCAACGTACGGAATTCCGAAC
1020
1030
ACCAATGCGTTCAGGCT
TGGTTACGCAAGTCCGA
1037