LOCUS Exported 7525 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS L2678 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7525) AUTHORS Fire A et al TITLE Fire Lab C. Elegans Vector Kit 1997 REFERENCE 2 (bases 1 to 7525) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7525 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 576..596 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" primer_bind complement(686..708) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(5099..5121) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 5221..5240 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(5318..5336) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5404..5508 /gene="bla" /label=AmpR promoter CDS 5509..6369 /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(5727..5746) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6540..7128 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7029..7048 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 7282..7299 /label=L4440 /note="L4440 vector, forward primer" protein_bind 7416..7437 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 7452..7482 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7490..7506 /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 7495..7517 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 7514..5 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 7514..5 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" ORIGIN 1 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tcaagagcat 61 ttgaatcaga atatggagaa cggagcatga gcattttcga agttttttag atgcactaga 121 acaaagcgtg ttggcttcct ctgagcccgc tttccttata tacccgcatt ctgcagcctt 181 acagaatgtt ctagaaggtc ctagatgcat tcgtttgaaa atactcccgg tgggtgcaaa 241 gagacgcaga cggaaaatgt atctgggtct ctttattgtg tacactactt ttccatgtac 301 cgaatgtgag tcgccctcct tttgcaacaa gcagctcgaa tgttctagaa aaaggtggaa 361 aatagtataa ataccgttga aaataaatac cgaacaacat ttgctctaat tgtgaaatta 421 gaaatcttca aactataatc atctcactgg atccccggga ttggccaaag gacccaaagg 481 tatgtttcga atgatactaa cataacatag aacattttca ggaggaccct tggctagagg 541 atccccgggt accgagctca gaaaaaatga ctgctccaaa gaagaagcgt aaggtaccgg 601 tgggtgaaga ccagaaacag cacctcgaac tgagccgcga tattgcccag cgtttcaacg 661 cgctgtatgg cgagatcgat cccgtcgttt tacaacgtcg tgactgggaa aaccctggcg 721 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag 781 aggcccgcac cgatcgccct tcccaacagt tgcgaaggta agtttaaaca gatccatact 841 aactaacttg ttctgacata attttcagct tgaatggcga atggcgcttt gcctggtttc 901 cggcaccaga agcggtgccg gaaagctggc tggagtgcga tcttcctgag gccgatactg 961 tcgtcgtccc ctcaaactgg cagatgcacg gttacgatgc gcccatctac accaacgtga 1021 cctatcccat tacggtcaat ccgccgtttg ttcccacgga gaatccgacg ggttgttact 1081 cgctcacatt taatgttgat gaaagctggc tacaggaagg ccagacgcga attatttttg 1141 atggcgttaa ctcggcgttt catctgtggt gcaacgggcg ctgggtcggt tacggccagg 1201 taagtttaat taagttgata ctaactaaca agatctgatt aattttcagg acagtcgttt 1261 gccgtctgaa tttgacctga gcgcattttt acgcgccgga gaaaaccgcc tcgcggtgat 1321 ggtgctgcgc tggagtgacg gcagttatct ggaagatcag gatatgtggc ggatgagcgg 1381 cattttccgt gacgtctcgt tgctgcataa accgactaca caaatcagcg atttccatgt 1441 tgccactcgc tttaatgatg atttctcccg cgctgtactg gaggctgaag tccaggtaag 1501 tttaaacagg atcttactaa ctaacatgct aacactgaat tttcagatgt gcggcgagtt 1561 gcgtgactac ctacgggtaa cagtttcttt atggcagggt gaaacgcagg tcgccagcgg 1621 caccgcgcct ttcggcggtg aaattatcga tgagcgtggt ggttatgccg atcgcgtcac 1681 actacgtctg aacgtcgaaa acccgaaact gtggagcgcc gaaatcccga atctctatcg 1741 tgcggtggtt gaactgcaca ccgccgacgg cacgctgatt gaagcagaag cctgcgatgt 1801 cggtttccgc gaggtgcgga ttgaaaatgg tctgctgctg ctgaacggca agccgttgct 1861 gattcgaggc gttaaccgtc acgagcatca tcctctgcat ggtcaggtca tggatgagca 1921 gacgatggtg caggatgtaa gtttaaacta ttcgttacta actaacttta aacatttaaa 1981 ttttcagatc ctgctgatga agcagaacaa ctttaacgcc gtgcgctgtt cgcattatcc 2041 gaaccatccg ctgtggtaca cgctgtgcga ccgctacggc ctgtatgtgg tggatgaagc 2101 caatattgaa acccacggca tggtgccaat gaatcgtctg accgatgatc cgcgctggct 2161 accggcgatg agcgaacgcg taacgcgaat ggtgcagcgc gatcgtaatc acccgagtgt 2221 gatcatctgg tcgctgggga atggtaagtt taaacagttg aatactaact aacggagatc 2281 tttgaaattt tcagaatcag gccacggcgc taatcacgac gcgctgtatc gctggatcaa 2341 atctgtcgat ccttcccgcc cggtgcagta tgaaggcggc ggagccgaca ccacggccac 2401 cgatattatt tgcccgatgt acgcgcgcgt ggatgaagac cagcccttcc cggctgtgcc 2461 gaaatggtcc atcaaaaaat ggctttcgct acctggagag acgcgcccgc tgattctttg 2521 cgaggtaagt ttaaacagaa ctctactaac taacacatta gatcctaatt ttcagtacgc 2581 tcacgcgatg ggcaacagtc ttggcggttt cgctaaatac tggcaggcgt ttcgtcagta 2641 tccccgttta cagggcggct tcgtctggga ctgggtggat cagtcgctga ttaaatatga 2701 tgaaaacggc aacccgtggt cggcttacgg cggtgatttt ggcgatacgc cgaacgatcg 2761 ccagttctgt atgaacggtc tggtctttgc cgaccgcacg ccgcatccag cgtaagttta 2821 aacaataacc tactaactaa cgtagataat ttaaattttc aggctgacgg aagcaaaaca 2881 ccagcagcag tttttccagt tccgtttatc cgggcaaacc atcgaagtga ccagcgaata 2941 cctgttccgt catagcgata acgagctcct gcactggatg gtggcgctgg atggtaagcc 3001 gctggcaagc ggtgaagtgc ctctggatgt cgctccacaa ggtaaacagt tgattgaact 3061 gcctgaacta ccgcagccgg agagcgccgg gcaactctgg ctcacagtac gcgtagtgca 3121 accgaacgcg accgcatggt cagaagccgg gcacatcagc gcatggcagc agtggaggta 3181 agtttaaaca agatcctact aactaactct acattgatga attttcagac tggcggaaaa 3241 cctcagtgtg acgctccccg ccgcgtccca cgccatcccg catctgacca ccagcgaaat 3301 ggatttttgc atcgagctgg gtaataagcg ttggcaattt aaccgccagt caggctttct 3361 ttcacagatg tggattggcg ataaaaaaca actgctgacg ccgctgcgcg atcagttcac 3421 ccgtgcaccg ctggataacg acattggcgt aagtgaagcg acccgcattg accctaacgc 3481 ctgggtcggt aagtttaaac aaagttgtac taactaacga agatcttgat aattttcaga 3541 acgctggaag gcggcgggcc attaccaggc cgaagcagcg ttgttgcagt gcacggcaga 3601 tacacttgct gatgcggtgc tgattacgac cgctcacgcg tggcagcatc aggggaaaac 3661 cttatttatc agccggaaaa cctaccggat tgatggtagt ggtcaaatgg cgattaccgt 3721 tgatgttgaa gtggcgagcg atacaccgca tccggcgcgg attggcctga actgccagct 3781 ggcgcaggta gcagagcggg taaactggct cggattaggg ccgcaagaaa actatcccga 3841 ccgccttact gccgcctgtt ttgaccgctg ggatctgcca ttgtcagaca tgtagtaagt 3901 ttaaacttga tagtactaac taacatgttt catttaaatt ttcagtaccc cgtacgtctt 3961 cccgagcgaa aacggtctgc gctgcgggac gcgcgaattg aattatggcc cacaccagtg 4021 gcgcggcgac ttccagttca acatcagccg ctacagtcaa cagcaactga tggaaaccag 4081 ccatcgccat ctgctgcacg cggaagaagg cacatggctg aatatcgacg gtttccatat 4141 ggggattggt ggcgacgact cctggagccc gtcagtatcg gcggaattcc aactgagcgc 4201 cggtcgctac cattaccaac ttgtctggtg tcaaaaataa taggggccgc tgtcatcaga 4261 gtaagtttaa actgagttct actaactaac gagtaatatt taaattttca gcatctcgcg 4321 cccgtgcctc tgacttctaa gtccaattac tcttcaacat ccctacatgc tctttctccc 4381 tgtgctccca ccccctattt ttgttattat caaaaaactt ctcttaattt ctttgttttt 4441 tagcttcttt taagtcacct ctaacaatga aattgtgtag attcaaaaat agaattaatt 4501 cgtaataaaa agtcgaaaaa aattgtgctc cctcccccca ttaataataa ttctatccca 4561 aaatctacac aatgttctgt gtacacttct tatgtttttt acttctgata aatttttttg 4621 aaacatcata gaaaaaaccg cacacaaaat accttatcat atgttacgtt tcagtttatg 4681 accgcaattt ttatttcttc gcacgtctgg gcctctcatg acgtcaaatc atgctcatcg 4741 tgaaaaagtt ttggagtatt tttggaattt ttcaatcaag tgaaagttta tgaaattaat 4801 tttcctgctt ttgctttttg gggtttcccc tattgtttgt caagatttcg aggacggcgt 4861 ttttcttgct aaaatcacaa gtattgatga gcacgatgca agaaagatcg gaagaaggtt 4921 tgggtttgag gctcagtgga aggtgagtag aagttgataa tttgaaagtg gagtagtgtc 4981 tatggggttt ttgccttaaa tgacagaata cattcccaat ataccaaaca taactgtttc 5041 ctactagtcg gccgtacggg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc 5101 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca 5161 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg 5221 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat 5281 gcgtaaggag aaaataccgc atcaggcggc cttaagggcc tcgtgatacg cctattttta 5341 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat 5401 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg 5461 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa 5521 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac 5581 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac 5641 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt 5701 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc 5761 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca 5821 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc 5881 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag 5941 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa 6001 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg 6061 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa 6121 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg 6181 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt 6241 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt 6301 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag 6361 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat 6421 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct 6481 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct 6541 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca 6601 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc 6661 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc 6721 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct 6781 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag 6841 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc 6901 tacaccgaac tgagatacct acagcgtgag cattgagaaa gcgccacgct tcccgaaggg 6961 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag 7021 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt 7081 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac 7141 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg 7201 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc 7261 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata 7321 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt 7381 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag 7441 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga 7501 taacaatttc acacaggaaa cagct //