LOCUS Exported 10549 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS BK48 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10549) AUTHORS Fire A et al TITLE Fire Lab C. Elegans Vector Kit 1997 REFERENCE 2 (bases 1 to 10549) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10549 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(1207..1235) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" primer_bind 1983..2003 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" primer_bind complement(7734..7753) /label=T7 /note="T7 promoter, forward primer" promoter complement(7735..7753) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(7760..7777) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(7760..7776) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7769..7791) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 7918..8373 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(8005..8024) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 8215..8236 /label=F1ori-F /note="F1 origin, forward primer" promoter 8400..8504 /gene="bla" /label=AmpR promoter CDS 8505..9365 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAISMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(8723..8742) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 9536..10124 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 10278..10295 /label=L4440 /note="L4440 vector, forward primer" promoter 10448..10478 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 10486..10502 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 10491..10513 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 10510..10526 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 10510..10526 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 10547..16 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" ORIGIN 1 taaccctcac taaagggaac aaaagctggc tccaaaatcg tcttccgctc tgaaaaacga 61 aagtggacct ttgacatccg aaaaaatggg cgaaaaaatg aaattgagct ttttgggtcg 121 aaaaaaatgt ttttagaatg ctgagaacac gttaaacacg aagatcatat ttattttgag 181 acccggatgc tctgaaaatg tctgacatag atttaaaaaa gcatatatat atttttcatt 241 ttcaacgtga aagttttgtg caactttata gaatctccta ttggcacatt gttttttatt 301 taactgaggc agtttttgaa cacctttttg aaactttgaa tctctttgaa gtatactgtc 361 gaaaagactg acttgagcgt tcgaaatgcc agaagaaaac tatatttgaa tctcgcgcta 421 aattgagaaa tgcaaccgcg ctccactgga caattggaaa aaaaatttat tcggaggcga 481 caacggtatt ttcgaaattg attttctgtg tattttctca ttttttataa attcttcttt 541 gatttatcgt tcgtttgtga gaaatttaat tgtattcaaa cttttttata gtaagatgtc 601 gtcgagtcgg tcacaatcac ctgaaactcc aaaggcagcc agtgaggaac gtgaagaaga 661 agaaaaagag tcatctgaac aggtttgatt ttctttctgg tcaaaaagat gaaattattg 721 attttcagcc agatactccc aaaactagca gcgagaagtc tgcaagtcgt tcacagtcgc 781 ccagagaatc gcgggaagtg agccaagagg tatgtttttc aaaaatcaat aactgatcat 841 aatttttatt gtttggtgaa tttaagaaaa taatattcga aaattcctct gaattatcaa 901 gattgcagta ttaatttcga gaaaaattga gatattcata gagctattgt aaattttctt 961 gatttcagac tgaaacttcg gaaaatcaag agaaaatcaa agaaaaggat gacggggatg 1021 atcagcctgg cacaccgaac agctatagaa gccgggaaac ttcaccagct ccaaaaaggt 1081 ccaaggagac caggtttgtc aaaagcttcc tgcgattaat tctcatttca atttttcaga 1141 gaatcagagt ctcctgaaaa atcaccggta gaaaaaatga gtaaaggaga agaacttttc 1201 actggagttg tcccaattct tgttgaatta gatggtgatg ttaatgggca caaattttct 1261 gtcagtggag agggtgaagg tgatgcaaca tacggaaaac ttacccttaa atttatttgc 1321 actactggaa aactacctgt tccatgggta agtttaaaca tatatatact aactaaccct 1381 gattatttaa attttcagcc aacacttgtc actactttct gttatggtgt tcaatgcttc 1441 tcgagatacc cagatcatat gaaacggcat gactttttca agagtgccat gcccgaaggt 1501 tatgtacagg aaagaactat atttttcaaa gatgacggga actacaagac acgtaagttt 1561 aaacagttcg gtactaacta accatacata tttaaatttt caggtgctga agtcaagttt 1621 gaaggtgata cccttgttaa tagaatcgag ttaaaaggta ttgattttaa agaagatgga 1681 aacattcttg gacacaaatt ggaatacaac tataactcac acaatgtata catcatggca 1741 gacaaacaaa agaatggaat caaagttgta agtttaaaca tgattttact aactaactaa 1801 tctgatttaa attttcagaa cttcaaaatt agacacaaca ttgaagatgg aagcgttcaa 1861 ctagcagacc attatcaaca aaatactcca attggcgatg gccctgtcct tttaccagac 1921 aaccattacc tgtccacaca atctgccctt tcgaaagatc ccaacgaaaa gagagaccac 1981 atggtccttc ttgagtttgt aacagctgct gggattacac atggcatgga tgaactatac 2041 aaatgcccgg ttcgttcaag atctcccaga aggtcttcag cacgttcccc gtcacgatct 2101 cctagacggc gccgagaaag aagctcagaa agaaagcaat ccgaagagcc agcaccgcta 2161 ccagagaaaa agaagaaaga gccgctggat attctacgaa caagaaccgg aggagcatat 2221 attccacccg ccaaacttcg acttatgcaa caacagatta gtgataagca aagtgaacag 2281 tatcagagaa tgaattggga aagaatgaag aaaaagattc acggattggt taacagagtc 2341 aacgcgaaga atcttgttca aattgtcaga gaacttcttc aagagaatgt gattcgttca 2401 aagtgagtga gaaaatcgaa ggaaaaggaa agaattaatt taatttttca ggggacttct 2461 ctgccgtgac attattcaag ctcaggcttt ctcaccagga ttctctaacg tctatgcagc 2521 tttggcggca gttatcaact cgaaattccc tcatgtcggt gaacttcttc tccgtcgtct 2581 gattgtacag ttcaaaagaa gtttccgtag aaatgacaga ggcgtcacgg tgaacgtgat 2641 caaattcatc gcacatttga ttaatcaaca agttgctcac gaagttcttg cgctggaaat 2701 catgattctg atgcttgaag aaccaactga tgattcagtt gaagtcgcca ttgcgttcct 2761 gaaagagtgt ggagcaaagc ttctggagat tgctccagca gctcttaaca gtgtctacga 2821 ccgtcttcgt gcaattctca tggaaactga aagatcggaa aatgcactgg atcgacgtat 2881 tcagtatatg attgagactg caatgcagat tcgaaaggac aaatttgcgg taaggtagaa 2941 tatataaata gtttattaga aaaaaataaa ttagaataat ttaaattcct actagccaat 3001 caggcgacct ttttgcgcat agttctatta ttgaaaaatt tggagaattt ctcatattct 3061 cgctcggaaa tctggaattc gacgagatct tctggcttct gtgcagctgc atcgctttgt 3121 gctccctttc tcgcttgtct tctgtgtaca ccaagaacct tgttgagttc atcaactgaa 3181 tctgtgactg gcttgttgct cactggatgc actagacgac tgattctcga gaaatcagat 3241 tgagttgcga ttagggtgac ctagaaattg ggaataatac gaacttttga aaatattcag 3301 gaggattaaa aaaattattc tcgacaatcc tacaaattta cttattgcac catgttgctc 3361 caacattttt cattaaaagt taatgaaaaa atgtagaaaa tcggaaattg gcaattttca 3421 gaccattttt aagcattttc aaaaaaaaat tgcagctgaa ataaatgtca ttttcagata 3481 aatcgagcga ttttctgttg tctgacacta gtttttagtt ttaaaaaatg ttggaagaac 3541 atggtgcaat aggtaatttc atagaatttc catgtgtttt ttttcaatta accaattatc 3601 caaatcttcc aaactcacat tttgcggagc tgggctatca agaatctgct gcagttttat 3661 aagacgagca tctctgatat cactgaaaat taatttttaa tcaaaacttg aatatcaact 3721 aaacccactt attaactttc tcgatcttct gtcgttcggt acgatgacgg tgaagaagcc 3781 aattgtagta gttgatttgg ttcaagtcct ttcggtgttg tacgtcagtg tcctgcaatg 3841 ctatttagtt ataacttagg cctaagattc aatttaatga agtgattaaa tttgttctct 3901 gaacctctta agatgatctt ttggattaga aacatataag acaggtttac ctatctatta 3961 aaaaacagat caaaatagat acgaccaaat cggataatcc atgcctacct ggcatctagg 4021 aacgtgttct tagaagattt cttacgtaat cgtatgaaga aataacaatt tgatcgttgg 4081 ccagcaaaaa tagggtttta agtgggatag tgtttttatt agctaaccgg aaaattttat 4141 agtttttttt tgcaagaaac cactgaaaac cccctaattg tatacatttt ttggagcagc 4201 ttctggtctt tttgagcaat aaaattcgat aaaacagaat ttaagtgtaa attgttcaca 4261 tttagtttct attttatcaa attttgttgc tcaaaaacat tcgaagctgc tctaaaaaaa 4321 tgcattaaaa aaggggtttt cagtggtttt tcacattaaa aaagctaatt ttaactaaaa 4381 atccatcata tttccaactt tgtcacaaca ataaaatgct ggtcaaaatg tgttcgaaaa 4441 aatgtttttt tttttaattt ttataattta aaaatagttt tctttcgctg ggacacatac 4501 atttttgggc gtaaattttc agttcaaatt tccattttta caaccataat cataaagcta 4561 cgtctgatct ctctcgcact tacctgcgcc tgattcgaaa gaacaaccgt agccaaaaga 4621 acaagaagaa caagcacgta gttgtggtag tggacgttca tcacgcaata ctgaccaatg 4681 gtcgtggggt ctcactttcc gtactattga gagaggggag actgaagatg gcaattgagg 4741 acagtgtctt cgacgcacgc atgcatccat aagcataatc caggagggat ggagagaaaa 4801 atcttgtttc taagcccctc cctttgtaat acatacacat atctaatacc gaagaatggc 4861 taattgaatg gacgtcagct gttgctgtag ttgccaaggc atcatcgatg aaataactga 4921 aagaaagaat taaataatta ttgcaggcgt atccggcggt cattgaagac ttggacttga 4981 ttgaggagga ggatcagatc atccatacac ttaatttgga ggatgcggtt gatccggaaa 5041 atgggcttag taagtgactg accacacgcg gggggcatta atttaataaa ttgaattcca 5101 tttcagatgt gttcaaacta gatccagaat tcgaaaagaa cgaggaggtt tatgaggaga 5161 tccgtaagga aatcattgga aacgccgata tttcggatga ggatggtggc gacgagttgg 5221 atgatgaaga agagggtagt gatgtggaag aggctccgaa gaagactaca gagattattg 5281 ataatactga tcagaattga ctgctttcag aaggtattca ttttgagttt tgggccggca 5341 aatctgtaag ttgccggttg ccgaaaattt gctgaatttg ccggaaaaaa aaattccgga 5401 atttatttaa aaactttttg taaaaattaa attaaatttg caacttttca gagaagtcta 5461 cctgacaatg caatcatctt tggactacca agaagctgct cacaaattgc tgaaaatgaa 5521 gattccagac agcatgcagg tcagcgatgt tgcaaagaaa aattttcgac caaaaaaacc 5581 aaccaatcat aaaatttaaa aaaaaactcc gtttttttct ttttttttat acgagaaaaa 5641 ccaaaaaaat gtatttttgc caaattctaa aatactatcc ccgaaatttt caatattttc 5701 tctttcagaa cgaactctgc gcgatgcttg tcgattgttg tgctcaacag cgtacctacg 5761 agcgattcta cggaatgctc atcgaacgtt tctgccgact tcgcctcgaa taccagcaat 5821 actttgaaaa gctctgccag gacacgtatt ccacgattca ccgaattgac atcacaaaac 5881 tgcggaattt ggctcgcctt attgctcatt tgctctcgac ggatgctatt gactggaaga 5941 ttttggccga tatgaaaatg accgaagagg acacaacttc ttctggcaga atctatatta 6001 aatatatatt taatgaactt gtggaggcga tgggaatggt taaacttcat tcgagagtta 6061 ctgatccgtg agtttcctag agagagttgt tttcgtattc aattttccct attttcagaa 6121 ctttggctca ttgctttgtt ggattattcc cacgaactaa tccgaacagc gcacgatttt 6181 cgatcaactt cttcacaatg attggattgg gtggtttgac gttggaactt cgtgaatggc 6241 tggcaaaggg tctcaagaag aagaagggaa tgctggatca gttgaaggcc gaatcaagct 6301 cagattcatc gtcgtcttcg gattcgtcag actcgtctga ttcttcggat tctgacgatt 6361 catccgactc gtcttcagat tcctcatctt cttcagaatc agagccagaa ccaccgaaga 6421 aaaagaagaa gaagaacagt gaagagagtt ccaaaaagaa ggaaaaagag aatattggtc 6481 gacgggatcg tggagacaag agagctgaac gtcatcgtga tcaaagtgtg gagaacaagg 6541 acaaggatcg tcgacgtcgc caggattctg acgaaaatcg tcggccagaa cgaggagatg 6601 accgcaagga tcggagtaaa gatcgtcgtc gtcaagactc ggatgatgag gatcggaaag 6661 gtcgtgaacg tcgggaagat tcaggggaaa gacgtcgcgg agatcgggat cgacgtgatc 6721 gaaacaagga tcaggaggat caccgtgaag atcgccgtga ccgaagcaag gatcgtgagg 6781 atcgacgtga tcgccgtcgt catgactctg atgatgatcg taaaactcgt cgggatagaa 6841 gtgaagagcg aggaggacgt cgtcgtgaag tggaatcgga tgatcgacgc cgacgtcgtt 6901 gaattttcaa attttaaata ctgaatattt gttttttttc ctattattta tttattctct 6961 ttgtgttttt tttcttgctt tctaaaaaat taattcaatc caaatctaaa catttttttt 7021 tctctttccg tctcccaatt cgtattccgc tcctctcatc tgaacacaat gtgcaagttt 7081 atttatcttc tcgctttcat ttcattagga cgtgggggga attggtggaa gggggaaaca 7141 cacaaaagga tgatggaaat gaaataagga cacacaatat gcaacaacat tcaattcaga 7201 aatatggagg aaggtttaaa agaaaacata aaaatatata gaggaggaag gaaaactagt 7261 aaaaaataag caaagaaatt aggcgaacga tgagaattgt cctcgcttgg caaatgcgaa 7321 tccgtatgga gaggcacgtt tggcgaaggc aaatgttcgg tatggagatc tgtaaaaatt 7381 tttaagttga aatttggtgt tgctctttta caaaattttc cgattttcgc ttgaaattac 7441 ggtgccaggt ctcgacacgt cttccaattt ttcaaattca aaagagcctt taatgggctg 7501 tagttgctaa tttctcgttt ttgaaaattt ttcttccgtt taatcgaaat ttgatgtatt 7561 ttatttatga ttttcaataa atttcaaaga aactggtgaa aactcggaaa attgtgaact 7621 acagtaatcc aatccttaaa ggcgcacacc ttttaaatgt ccgccccaat acgatatttt 7681 tttaagattc gggccctcta gagcggccgc caccgcggtg gagctccaat tcgccctata 7741 gtgagtcgta ttacaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg 7801 gcgttaccca acttaatcgc cttgcagcac atcccccctt cgccagctgg cgtaatagcg 7861 aagaggcccg caccgatcgc ccttcccaac agttgcgtag cctgaatggc gaatgggacg 7921 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta 7981 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt 8041 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg 8101 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat 8161 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac 8221 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag 8281 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg 8341 cgaattttaa caaaatatta acgtttacaa tttcaggtgg cacttttcgg ggaaatgtgc 8401 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac 8461 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt 8521 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag 8581 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg 8641 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa 8701 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc 8761 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag 8821 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa 8881 gcatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc 8941 taaccgcttt ttttcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg 9001 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa 9061 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa 9121 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg 9181 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag 9241 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg ggcagtcagg 9301 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt 9361 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt 9421 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac 9481 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag 9541 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg 9601 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca 9661 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga 9721 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca 9781 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc 9841 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca 9901 ccgaactgag atacctacag cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa 9961 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc 10021 caggggggaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc 10081 gtcgattttt gtgatgctcg tcaggggggc cgagcctatg gaaaaacgcc agcaacgcgg 10141 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat 10201 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca 10261 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca 10321 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg 10381 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttacctcact cattaggcac 10441 cccaggcttt acactttatg cttccggctc ctatgttgtg tggaattgtg agcggataac 10501 aatttcacac aggaaacagc tatgaccatg attacgccaa gctcggaat //