LOCUS Exported 8414 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS L2685 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8414) AUTHORS Fire A et al TITLE Fire Lab C. Elegans Vector Kit 1997 REFERENCE 2 (bases 1 to 8414) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8414 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 583..603 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" primer_bind complement(646..674) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" primer_bind 1422..1442 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" primer_bind complement(1575..1597) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(5988..6010) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 6110..6129 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(6207..6225) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 6293..6397 /gene="bla" /label=AmpR promoter CDS 6398..7258 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(6616..6635) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7429..8017 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7918..7937 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 8171..8188 /label=L4440 /note="L4440 vector, forward primer" protein_bind 8305..8326 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 8341..8371 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8379..8395 /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 8384..8406 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 8403..5 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 8403..5 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" ORIGIN 1 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tcaccaaaaa 61 cggaacgttg agctggacgg aaatagtggt aaagtgacat gattatagtt tgaagatttc 121 taatttcaca attagagcaa atgttgttcg gtatttattt tcaacggtat ttatactatt 181 ttccaccttt ttctagaaca ttcgagctgc ttgttgcaaa aggagggcga ctcacattcg 241 gtacatggaa aagtagtgta cacaataaag agacccagat acattttccg tctgcgtctc 301 tttgcaccca ccgggagtat tttcaaacga atgcatctag gaccttctag aacattctgt 361 aaggctgcag aatgcgggta tataaggaaa gcgggctcag aggaagccaa cacgctttgt 421 tctagtgcat ctaaaaaact tcgaaaatcc tcatcggatc cccgggattg gccaaaggac 481 ccaaaggtat gtttcgaatg atactaacat aacatagaac attttcagga ggacccttgg 541 ctagaggatc cccgggtacc gagctcagaa aaaatgactg ctccaaagaa gaagcgtaag 601 gtaccggtag aaaaaatgag taaaggagaa gaacttttca ctggagttgt cccaattctt 661 gttgaattag atggtgatgt taatgggcac aaattttctg tcagtggaga gggtgaaggt 721 gatgcaacat acggaaaact tacccttaaa tttatttgca ctactggaaa actacctgtt 781 ccatgggtaa gtttaaacat atatatacta actaaccctg attatttaaa ttttcagcca 841 acacttgtca ctactttctg ttatggtgtt caatgcttct cgagataccc agatcatatg 901 aaacggcatg actttttcaa gagtgccatg cccgaaggtt atgtacagga aagaactata 961 tttttcaaag atgacgggaa ctacaagaca cgtaagttta aacagttcgg tactaactaa 1021 ccatacatat ttaaattttc aggtgctgaa gtcaagtttg aaggtgatac ccttgttaat 1081 agaatcgagt taaaaggtat tgattttaaa gaagatggaa acattcttgg acacaaattg 1141 gaatacaact ataactcaca caatgtatac atcatggcag acaaacaaaa gaatggaatc 1201 aaagttgtaa gtttaaacat gattttacta actaactaat ctgatttaaa ttttcagaac 1261 ttcaaaatta gacacaacat tgaagatgga agcgttcaac tagcagacca ttatcaacaa 1321 aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa 1381 tctgcccttt cgaaagatcc caacgaaaag agagaccaca tggtccttct tgagtttgta 1441 acagctgctg ggattacaca tggcatggat gaactataca aatgcccggt gggtgaagac 1501 cagaaacagc acctcgaact gagccgcgat attgcccagc gtttcaacgc gctgtatggc 1561 gagatcgatc ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt 1621 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc 1681 gatcgccctt cccaacagtt gcgaaggtaa gtttaaacag atccatacta actaacttgt 1741 tctgacataa ttttcagctt gaatggcgaa tggcgctttg cctggtttcc ggcaccagaa 1801 gcggtgccgg aaagctggct ggagtgcgat cttcctgagg ccgatactgt cgtcgtcccc 1861 tcaaactggc agatgcacgg ttacgatgcg cccatctaca ccaacgtgac ctatcccatt 1921 acggtcaatc cgccgtttgt tcccacggag aatccgacgg gttgttactc gctcacattt 1981 aatgttgatg aaagctggct acaggaaggc cagacgcgaa ttatttttga tggcgttaac 2041 tcggcgtttc atctgtggtg caacgggcgc tgggtcggtt acggccaggt aagtttaatt 2101 aagttgatac taactaacaa gatctgatta attttcagga cagtcgtttg ccgtctgaat 2161 ttgacctgag cgcattttta cgcgccggag aaaaccgcct cgcggtgatg gtgctgcgct 2221 ggagtgacgg cagttatctg gaagatcagg atatgtggcg gatgagcggc attttccgtg 2281 acgtctcgtt gctgcataaa ccgactacac aaatcagcga tttccatgtt gccactcgct 2341 ttaatgatga tttctcccgc gctgtactgg aggctgaagt ccaggtaagt ttaaacagga 2401 tcttactaac taacatgcta acactgaatt ttcagatgtg cggcgagttg cgtgactacc 2461 tacgggtaac agtttcttta tggcagggtg aaacgcaggt cgccagcggc accgcgcctt 2521 tcggcggtga aattatcgat gagcgtggtg gttatgccga tcgcgtcaca ctacgtctga 2581 acgtcgaaaa cccgaaactg tggagcgccg aaatcccgaa tctctatcgt gcggtggttg 2641 aactgcacac cgccgacggc acgctgattg aagcagaagc ctgcgatgtc ggtttccgcg 2701 aggtgcggat tgaaaatggt ctgctgctgc tgaacggcaa gccgttgctg attcgaggcg 2761 ttaaccgtca cgagcatcat cctctgcatg gtcaggtcat ggatgagcag acgatggtgc 2821 aggatgtaag tttaaactat tcgttactaa ctaactttaa acatttaaat tttcagatcc 2881 tgctgatgaa gcagaacaac tttaacgccg tgcgctgttc gcattatccg aaccatccgc 2941 tgtggtacac gctgtgcgac cgctacggcc tgtatgtggt ggatgaagcc aatattgaaa 3001 cccacggcat ggtgccaatg aatcgtctga ccgatgatcc gcgctggcta ccggcgatga 3061 gcgaacgcgt aacgcgaatg gtgcagcgcg atcgtaatca cccgagtgtg atcatctggt 3121 cgctggggaa tggtaagttt aaacagttga atactaacta acggagatct ttgaaatttt 3181 cagaatcagg ccacggcgct aatcacgacg cgctgtatcg ctggatcaaa tctgtcgatc 3241 cttcccgccc ggtgcagtat gaaggcggcg gagccgacac cacggccacc gatattattt 3301 gcccgatgta cgcgcgcgtg gatgaagacc agcccttccc ggctgtgccg aaatggtcca 3361 tcaaaaaatg gctttcgcta cctggagaga cgcgcccgct gattctttgc gaggtaagtt 3421 taaacagaac tctactaact aacacattag atcctaattt tcagtacgct cacgcgatgg 3481 gcaacagtct tggcggtttc gctaaatact ggcaggcgtt tcgtcagtat ccccgtttac 3541 agggcggctt cgtctgggac tgggtggatc agtcgctgat taaatatgat gaaaacggca 3601 acccgtggtc ggcttacggc ggtgattttg gcgatacgcc gaacgatcgc cagttctgta 3661 tgaacggtct ggtctttgcc gaccgcacgc cgcatccagc gtaagtttaa acaataacct 3721 actaactaac gtagataatt taaattttca ggctgacgga agcaaaacac cagcagcagt 3781 ttttccagtt ccgtttatcc gggcaaacca tcgaagtgac cagcgaatac ctgttccgtc 3841 atagcgataa cgagctcctg cactggatgg tggcgctgga tggtaagccg ctggcaagcg 3901 gtgaagtgcc tctggatgtc gctccacaag gtaaacagtt gattgaactg cctgaactac 3961 cgcagccgga gagcgccggg caactctggc tcacagtacg cgtagtgcaa ccgaacgcga 4021 ccgcatggtc agaagccggg cacatcagcg catggcagca gtggaggtaa gtttaaacaa 4081 gatcctacta actaactcta cattgatgaa ttttcagact ggcggaaaac ctcagtgtga 4141 cgctccccgc cgcgtcccac gccatcccgc atctgaccac cagcgaaatg gatttttgca 4201 tcgagctggg taataagcgt tggcaattta accgccagtc aggctttctt tcacagatgt 4261 ggattggcga taaaaaacaa ctgctgacgc cgctgcgcga tcagttcacc cgtgcaccgc 4321 tggataacga cattggcgta agtgaagcga cccgcattga ccctaacgcc tgggtcggta 4381 agtttaaaca aagttgtact aactaacgaa gatcttgata attttcagaa cgctggaagg 4441 cggcgggcca ttaccaggcc gaagcagcgt tgttgcagtg cacggcagat acacttgctg 4501 atgcggtgct gattacgacc gctcacgcgt ggcagcatca ggggaaaacc ttatttatca 4561 gccggaaaac ctaccggatt gatggtagtg gtcaaatggc gattaccgtt gatgttgaag 4621 tggcgagcga tacaccgcat ccggcgcgga ttggcctgaa ctgccagctg gcgcaggtag 4681 cagagcgggt aaactggctc ggattagggc cgcaagaaaa ctatcccgac cgccttactg 4741 ccgcctgttt tgaccgctgg gatctgccat tgtcagacat gtagtaagtt taaacttgat 4801 agtactaact aacatgtttc atttaaattt tcagtacccc gtacgtcttc ccgagcgaaa 4861 acggtctgcg ctgcgggacg cgcgaattga attatggccc acaccagtgg cgcggcgact 4921 tccagttcaa catcagccgc tacagtcaac agcaactgat ggaaaccagc catcgccatc 4981 tgctgcacgc ggaagaaggc acatggctga atatcgacgg tttccatatg gggattggtg 5041 gcgacgactc ctggagcccg tcagtatcgg cggaattcca actgagcgcc ggtcgctacc 5101 attaccaact tgtctggtgt caaaaataat aggggccgct gtcatcagag taagtttaaa 5161 ctgagttcta ctaactaacg agtaatattt aaattttcag catctcgcgc ccgtgcctct 5221 gacttctaag tccaattact cttcaacatc cctacatgct ctttctccct gtgctcccac 5281 cccctatttt tgttattatc aaaaaacttc tcttaatttc tttgtttttt agcttctttt 5341 aagtcacctc taacaatgaa attgtgtaga ttcaaaaata gaattaattc gtaataaaaa 5401 gtcgaaaaaa attgtgctcc ctccccccat taataataat tctatcccaa aatctacaca 5461 atgttctgtg tacacttctt atgtttttta cttctgataa atttttttga aacatcatag 5521 aaaaaaccgc acacaaaata ccttatcata tgttacgttt cagtttatga ccgcaatttt 5581 tatttcttcg cacgtctggg cctctcatga cgtcaaatca tgctcatcgt gaaaaagttt 5641 tggagtattt ttggaatttt tcaatcaagt gaaagtttat gaaattaatt ttcctgcttt 5701 tgctttttgg ggtttcccct attgtttgtc aagatttcga ggacggcgtt tttcttgcta 5761 aaatcacaag tattgatgag cacgatgcaa gaaagatcgg aagaaggttt gggtttgagg 5821 ctcagtggaa ggtgagtaga agttgataat ttgaaagtgg agtagtgtct atggggtttt 5881 tgccttaaat gacagaatac attcccaata taccaaacat aactgtttcc tactagtcgg 5941 ccgtacgggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg 6001 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt 6061 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag 6121 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga 6181 aaataccgca tcaggcggcc ttaagggcct cgtgatacgc ctatttttat aggttaatgt 6241 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac 6301 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc 6361 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt 6421 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct 6481 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga 6541 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag 6601 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca 6661 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga 6721 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag 6781 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc 6841 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa 6901 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt 6961 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg 7021 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt 7081 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg 7141 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat 7201 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact 7261 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa 7321 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt 7381 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt 7441 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg 7501 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca 7561 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt 7621 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga 7681 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc 7741 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact 7801 gagataccta cagcgtgagc attgagaaag cgccacgctt cccgaaggga gaaaggcgga 7861 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg 7921 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt 7981 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt 8041 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga 8101 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac 8161 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc 8221 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa 8281 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc 8341 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca 8401 cacaggaaac agct //