LOCUS Gateway_Nova_pCOLA-3-DEST 5134 bp DNA circular 24-AUG-2007
DEFINITION Gateway Nova pCOLA-3-DEST Strep-Tag II/attR1-CamccdB-attR2/6xHis-Tag.
SOURCE
ORGANISM
FEATURES Location/Qualifiers
CDS complement(224..1039)
/vntifkey="4"
/label=Kan
CDS complement(1982..3064)
/vntifkey="4"
/label=lacI
/note="Wildtype lacI gene"
promoter 3188..3204
/vntifkey="30"
/label=T7\Promoter
protein_bind 3209..3229
/vntifkey="31"
/label=lacO
RBS 3262..3267
/vntifkey="32"
/label=RBS-1
primer complement(5067..5085)
/vntifkey="27"
/label=T7\Terminator\primer
terminator 5081..5128
/vntifkey="43"
/label=T7\Terminator
RBS complement(1045..1049)
/vntifkey="32"
/label=RBS
-35_signal complement(1104..1109)
/vntifkey="48"
/label=-35\Ap
-10_signal complement(1081..1087)
/vntifkey="47"
/label=-10\Ap
rep_origin 1149..1784
/vntifkey="33"
/label=ColA\ori
-10_signal complement(3098..3103)
/vntifkey="47"
/label=lacI\-10
-35_signal complement(3122..3127)
/vntifkey="48"
/label=lacI\-35
protein_bind 3284..3307
/vntifkey="31"
/label=Strep-Tag\II
misc_recomb 3317..3441
/vntifkey="86"
/label=attR1
mutation 4065..4065
/vntifkey="62"
/label=C\to\T\-NcoI
mutation 4769..4769
/vntifkey="62"
/label=C\to\T\-BsaI
CDS 3550..4209
/vntifkey="4"
/label=Cam
CDS 4551..4856
/vntifkey="4"
/label=ccdB
misc_binding 5023..5040
/vntifkey="20"
/label=6x\His-Tag
primer 3274..3313
/vntifkey="27"
/label=S-NT\Strep\GW
primer 5017..5052
/vntifkey="27"
/label=S-CT\6xHis\GW
misc_recomb complement(4897..5021)
/vntifkey="86"
/label=attR2
misc_feature 3325..5006
/vntifkey="21"
/label=Region\replaced\by\pENTR\target
primer 3188..3207
/vntifkey="27"
/label=T7\Promoter\Primer
BASE COUNT 1364 a 1246 c 1209 g 1315 t
ORIGIN
1 cctcaggcat ttgagaagca cacggtcaca ctgcttccgg tagtcaataa accggtaaac
61 cagcaataga cataagcggc tatttaacga ccctgccctg aaccgacgac aagctgacga
121 ccgggtctcc gcaagtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
181 tttctaaata cattcaaata tgtatccgct catgaattaa ttcttagaaa aactcatcga
241 gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
301 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
361 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
421 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
481 gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
541 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa
601 atacgcggtc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
661 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
721 atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
781 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat
841 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
901 gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
961 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctagag caagacgttt
1021 cccgttgaat atggctcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
1081 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggcatgc
1141 tagcgcagaa acgtcctaga agatgccagg aggatactta gcagagagac aataaggccg
1201 gagcgaagcc gtttttccat aggctccgcc cccctgacga acatcacgaa atctgacgct
1261 caaatcagtg gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctgatg
1321 gctccctctt gcgctctcct gttcccgtcc tgcggcgtcc gtgttgtggt ggaggcttta
1381 cccaaatcac cacgtcccgt tccgtgtaga cagttcgctc caagctgggc tgtgtgcaag
1441 aaccccccgt tcagcccgac tgctgcgcct tatccggtaa ctatcatctt gagtccaacc
1501 cggaaagaca cgacaaaacg ccactggcag cagccattgg taactgagaa ttagtggatt
1561 tagatatcga gagtcttgaa gtggtggcct aacagaggct acactgaaag gacagtattt
1621 ggtatctgcg ctccactaaa gccagttacc aggttaagca gttccccaac tgacttaacc
1681 ttcgatcaaa ccgcctcccc aggcggtttt ttcgtttaca gagcaggaga ttacgacgat
1741 cgtaaaagga tctcaagaag atcctttacg gattcccgac accatcactc tagatttcag
1801 tgcaatttat ctcttcaaat gtagcacctg aagtcagccc catacgatat aagttgtaat
1861 tctcatgtta gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa
1921 gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg
1981 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
2041 acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg
2101 agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt
2161 ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat
2221 aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatgtccgca ccaacgcgca
2281 gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca
2341 tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg
2401 cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat
2461 gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga
2521 tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg
2581 agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat
2641 tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca
2701 gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc
2761 ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg
2821 ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca
2881 acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca
2941 tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc
3001 gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt
3061 tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg
3121 ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt
3181 aggaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctgtagaa
3241 ataattttgt ttaactttaa taaggagata taccatggca agctggagcc acccgcagtt
3301 cgaaaagggt gcagtcacaa gtttgtacaa aaaagctgaa cgagaaacgt aaaatgatat
3361 aaatatcaat atattaaatt agattttgca taaaaaacag actacataat actgtaaaac
3421 acaacatatc cagtcactat ggcggccgca ttaggcaccc caggctttac actttatgct
3481 tccggctcgt ataatgtgtg gattttgagt taggatccgt cgagattttc aggagctaag
3541 gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat
3601 cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt
3661 cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg
3721 gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaattccg tatggcaatg
3781 aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag
3841 caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta
3901 cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg
3961 tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat
4021 ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcactatggg caaatattat
4081 acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat
4141 ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc
4201 ggggcgtaaa cgcgtggatc cggcttacta aaagccagat aacagtatgc gtatttgcgc
4261 gctgattttt gcggtataag aatatatact gatatgtata cccgaagtat gtcaaaaaga
4321 ggtatgctat gaagcagcgt attacagtga cagttgacag cgacagctat cagttgctca
4381 aggcatatat gatgtcaata tctccggtct ggtaagcaca accatgcaga atgaagcccg
4441 tcgtctgcgt gccgaacgct ggaaagcgga aaatcaggaa gggatggctg aggtcgcccg
4501 gtttattgaa atgaacggct cttttgctga cgagaacagg ggctggtgaa atgcagttta
4561 aggtttacac ctataaaaga gagagccgtt atcgtctgtt tgtggatgta cagagtgata
4621 ttattgacac gcccgggcga cggatggtga tccccctggc cagtgcacgt ctgctgtcag
4681 ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga
4741 tgaccaccga tatggccagt gtgccggttt ccgttatcgg ggaagaagtg gctgatctca
4801 gccaccgcga aaatgacatc aaaaacgcca ttaacctgat gttctgggga atataaatgt
4861 caggctccct tatacacagc cagtctgcag gtcgaccata gtgactggat atgttgtgtt
4921 ttacagtatt atgtagtctg ttttttatgc aaaatctaat ttaatatatt gatatttata
4981 tcattttacg tttctcgttc agctttcttg tacaaagtgg ttcatcacca ccatcatcac
5041 taagtgatta acctaggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
5101 ctctaaacgg gtcttgaggg gttttttgct gaaa
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