LOCUS Exported 6359 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCG150 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6359) TITLE Seydoux Lab - Third Generation Vector Kit REFERENCE 2 (bases 1 to 6359) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6359 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 168..184 /label=pBluescriptKS /note="For pBluescript vector" primer_bind 169..185 /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind 2542..2564 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(2602..2620) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 2688..2792 /gene="bla" /label=AmpR promoter CDS 2793..3653 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGSDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(3011..3030) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 3824..4412 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4313..4332 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4539..4561 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 4558..4574 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 4558..4574 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" protein_bind 4593..4717 /gene="mutant version of attR" /label=attR4 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS complement(4758..5063) /codon_start=1 /gene="ccdB" /product="CcdB, a bacterial toxin that poisons DNA gyrase" /label=ccdB /note="Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains." /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" primer_bind complement(4860..4879) /label=ccdB-fwd /note="ccdB gene, forward primer" CDS complement(5383..6063) /codon_start=1 /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE DPAY" primer_bind 5978..5997 /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" promoter complement(6117..6147) /label=lac UV5 promoter /note="E. coli lac promoter with an ""up"" mutation" protein_bind complement(6172..6295) /gene="mutant version of attR" /label=attR3 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(6303..6320) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(6303..6319) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6312..6334) /label=M13/pUC Forward /note="In lacZ gene" ORIGIN 1 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga 61 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct 121 ccttacgcat ctgtgcggta tttcacaccg catatgccgg gcccccctcg aggtcgacgg 181 tatcgataag cttcagtaaa agaagtagaa ttttatagtt ttttttctgt ttgaaaaatt 241 ctccccatca atgttctttc aaataaatac atcactaatg caaagtattc tataacctca 301 tatctaaatt cttcaaaatc ttaacatatc ttatcattgc tttaagtcaa acgtaacatt 361 aaaaaaaatg ttttggaaaa tgtgtcaagt ctctcaaaat tcagtttttt aaaccactcc 421 tatagtccta tagtcctata gttacccatg aaatccttat atattactgt aaaatgtttc 481 aaaaaccatt ggcaaattgc cagaactgaa aatttccggc aaattgggga accggcaaat 541 tgccaatttg ctgaatttgc cggaaaaacg gtaattgccg aaagtttttg acacgaaaat 601 ggcaaattgt ggttttaaaa tttttttttt tggaaatttc agaatttcaa ttttaatcgg 661 caaaactgta ggcatcctaa gaatgttcct acatctattt tgaaaagtaa gcgaattcta 721 tgaaaatgtc taaagaaaat ggggaaacaa tttcaaaaag gcacagtttc aagtgtttcc 781 gaattatact aaatccctct aaaaacttcc ggcaaattga tatccgtaaa agagcaaatc 841 cgcatttttg ccgaaaatta aaatttccga caaatcggca aaccggcaat ttggcgaaat 901 ttgccggaag gcaattgccg cccacccctg ttccagaggt tcaaactggt agcaaagctc 961 aaaatttctc aaattctcca attttttttt gaattttggc agtgtaccaa aatgacattc 1021 agtcatattg gtttattata gatttattta gataaaatcc taaatgattc tacctttaaa 1081 gatgcccact ttaaaagtaa tgactcaaac ttcaaattgc tctaagattc tattgaatta 1141 ccatcttttc ctctcatttt ctctcactgt ctatttcatc acaaattcat ccctctctcc 1201 ctctctttct ctctccctct ctctctttct ctttgctcat catctgtcat tttgtccgtt 1261 cctctctctg cgccctcagc gttccccaca ctctctcgct tctcttttcc tagacgtctt 1321 cttttttcat cttcttcagc cctttttcgc cattttccat ctctgtcaat cattacggac 1381 gacccccatt aatttttgga tgtgctctgc gagcaagagc acacggaact catttatttt 1441 ttgttggaat ttctttttca ttttagttag ttaaaagcta caacagcata tatgaaggca 1501 gagcaacaac aacaatcgat cgcacccggc tcggcaacct tcccgtctca gatgccccga 1561 cccccaccgg taaccgaaca ggctataacc accgaggcgg agcttctcgc gaaaaatcaa 1621 attacaccaa atgatgtgct agctcttcca ggaatcactc aaggattcct atgctcccca 1681 tcggcaaacg tctataacat tgagttcacc aaattccaaa tccgtgacct cgacacggaa 1741 cacgtgctct tcgaaattgc aaaaccggaa aatgagacgg aagagaatct gcaggcgcaa 1801 gccgaatcgg caagatatgt ccgatatcga tttgcgccga attttctgaa attaaagacg 1861 gtcggcgcga cggtcgagtt caaggtcggc gatgtgccga tcacacattt tcgaatgatt 1921 gaacggcact ttttcaagga tcgtttattg aaatgctttg attttgaatt tggattctgt 1981 atgccaaatt cgagaaacaa ctgtgaacat atctatgaat ttccacaact ttcacaacaa 2041 ctcatggacg acatgatcaa taatccaaac gagacccgct ccgatagctt ctattttgtg 2101 gagaataagc tcgtaatgca caataaggcc gactactcgt atgatgcata atttcccgcc 2161 gatggaataa aactaataat ttttctcgca atttatagaa atcatgtgat tttcctaccg 2221 ctaaatttaa aatttcctaa tttttcacta tatttctcac aaaatgcatt ccgaaaactc 2281 tcatttttca gcttttcaat acaaatccca ataaatgatt tttcaaatat aaaaaaaaaa 2341 acggaattcc tgcagcccgg gggatccgct agttctagag cggccgccac cgcggtggag 2401 ctcccatatg gtgcactctc agtgcaatct gctctgatgc cgcatagtta agccagcccc 2461 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt 2521 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac 2581 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga 2641 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta 2701 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat 2761 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc 2821 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga 2881 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca 2941 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt 3001 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg 3061 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc 3121 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata 3181 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt 3241 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag 3301 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca 3361 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg 3421 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg 3481 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggtcag 3541 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg 3601 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag 3661 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga 3721 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt 3781 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc 3841 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc 3901 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac 3961 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac 4021 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt 4081 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct 4141 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat 4201 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt 4261 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg 4321 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt 4381 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt 4441 tcctggtctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg 4501 tggataaccg tattaccgcc tttgagtgag ctgataccag cggataacaa tttcacacag 4561 gaaacagcta tgaccatgat tacgccaagc tatcaacttt gtatagaaaa gttgaacgag 4621 aaacgtaaaa tgatataaat atcaatatat taaattagat tttgcataaa aaacagacta 4681 cataatactg taaaacacaa catatccagt cactatggtc gacctgcaga ctggctgtgt 4741 ataagggagc ctgacattta tattccccag aacatcaggt taatggcgtt tttgatgtca 4801 ttttcgcggt ggctgagatc agccacttct tccccgataa cggagaccgg cacactggcc 4861 atatcggtgg tcatcatgcg ccagctttca tccccgatat gcaccaccgg gtaaagttca 4921 cgggggactt tatctgacag cagacgtgca ctggccaggg ggatcaccat ccgtcgcccg 4981 ggcgtgtcaa taatatcact ctgtacatcc acaaacagac gataacggct ctctctttta 5041 taggtgtaaa ccttaaactg catttcacca gcccctgttc tcgtcggcaa aagagccgtt 5101 catttcaata aaccgggcga cctcagccat cccttcctga ttttccgctt tccagcgttc 5161 ggcacgcaga cgacgggctt cattctgcat ggttgtgctt accgaaccgg agatattgac 5221 atcatatatg ccttgagcaa ctgatagctg tcgctgtcaa ctgtcactgt aatacgctgc 5281 ttcatagcat acctcttttt gacatacttc gggtatacat atcagtatat attcttatac 5341 cgcaaaaatc agcgcgcaaa tacgcatact gttatctggc ttttagtaag ccggatcctc 5401 tagattacgc cccgcctgcc actcatcgca gtactgttgt aattcattaa gcattctgcc 5461 gacatggaag ccatcacaaa cggcatgatg aacctgaatc gccagcggca tcagcacctt 5521 gtcgccttgc gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt 5581 ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaga cgaaaaacat 5641 attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg 5701 cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa 5761 cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag 5821 ctcaccgtct ttcattgcca tacggaattc cggatgagca ttcatcaggc gggcaagaat 5881 gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt 5941 aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa 6001 atgttcttta cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc 6061 cattttagct tccttagctc ctgaaaatct cgacggatcc taactcaaaa tccacacatt 6121 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgcggccg ccatagtgac 6181 tggatatgtt gtgttttaca gtattatgta gtctgttttt tatgcaaaat ctaatttaat 6241 atattgatat ttatatcatt ttacgtttct cgttcaactt tattatacat agttgataat 6301 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaa //