LOCUS Exported 2827 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS TAL021 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2827) AUTHORS Sander JD, Cade L, Khayter C, Reyon D, Peterson RT, Joung JK, Yeh JR TITLE Targeted gene disruption in somatic zebrafish cells using engineered TALENs. JOURNAL Nat Biotechnol. 2011 Aug 5;29(8):697-8. doi: 10.1038/nbt.1934. PUBMED 21822241 REFERENCE 2 (bases 1 to 2827) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2827 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(29..51) /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(146..610) /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITPSLHAGLCSRRARDPVSSVHDPGTVQAAGAVLFPAPAYRQ YWRRWRPLVQASIVFSRCIREVPSSNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRN SEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICSDAA" primer_bind 151..170 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 364..386 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 378..395 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 379..395 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(606..622) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(606..622) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(619..641) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 630..646 /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)." promoter complement(654..684) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 699..720 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(837..854) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1008..1596) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1088..1107) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1767..2627) /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 2390..2409 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2628..2732) /gene="bla" /label=AmpR promoter primer_bind 2800..2818 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acctcgcgaa 421 tgcatctaga gaagacaatt gacgcctgca caagtggtcg ccatcgcctc caatattggc 481 ggtaagcagg cgctggaaac agtacagcgc ctgctgcctg tactgtgcca ggatcatgga 541 ctgaagagac cggatcccgg gcccgtcgac tgcagaggcc tgcatgcaag cttggcgtaa 601 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata 661 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta 721 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa 781 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg 841 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag 901 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa 961 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc 1021 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca 1081 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg 1141 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct 1201 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt 1261 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag 1321 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc 1381 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac 1441 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga 1501 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc 1561 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg 1621 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca 1681 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt 1741 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca 1801 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg 1861 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga gccacgctca 1921 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt 1981 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt 2041 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca 2101 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca 2161 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga 2221 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact 2281 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga 2341 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg 2401 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc 2461 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga 2521 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat 2581 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt 2641 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt 2701 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac 2761 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc 2821 tttcgtc //