LOCUS Exported 2831 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS TAL006 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2831) 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 2831) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2831 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(29..51) /label=pGEX 3' /note="pGEX vectors, reverse primer" 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(610..626) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(610..626) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(623..645) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 634..650 /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(658..688) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 703..724 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(841..858) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1012..1600) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1092..1111) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1771..2631) /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 2394..2413 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2632..2736) /gene="bla" /label=AmpR promoter primer_bind 2804..2822 /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 gaagacaaga acctgacccc agaccaggta gtcgcaatcg cgtcgaacat 481 tgggggaaag caagccctgg aaaccgtgca aaggttgttg ccggtccttt gtcaagacca 541 cggccttaag agaccggatc ccgggcccgt cgactgcaga ggcctgcatg caagcttggc 601 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa 661 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac 721 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca 781 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc 841 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc 901 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc 961 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 1021 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 1081 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 1141 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 1201 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 1261 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 1321 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 1381 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 1441 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 1501 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt 1561 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc 1621 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt 1681 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta 1741 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat 1801 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac 1861 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagagccacg 1921 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag 1981 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt 2041 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt 2101 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt 2161 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt 2221 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct 2281 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt 2341 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac 2401 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa 2461 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa 2521 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca 2581 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct 2641 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga 2701 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc 2761 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag 2821 gccctttcgt c //