LOCUS Exported 3001 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pFUS_A2A SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3001) AUTHORS Sakuma T, Hosoi S, Woltjen K, Suzuki KI, Kashiwagi K, Wada H, Ochiai H, Miyamoto T, Kawai N, Sasakura Y, Matsuura S, Okada Y, Kawahara A, Hayashi S, Yamamoto T TITLE Efficient TALEN construction and evaluation methods for human cell and animal applications. JOURNAL Genes Cells. 2013 Feb 6. doi: 10.1111/gtc.12037. PUBMED 23388034 REFERENCE 2 (bases 1 to 3001) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3001 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 413..1204 /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=SmR /note="confers resistance to spectinomycin and streptomycin" /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" rep_origin 1297..1885 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1786..1805 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2039..2056 /label=L4440 /note="L4440 vector, forward primer" terminator 2215..2242 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 2334..2420 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" CDS complement(2474..2851) /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGLDSH HLVRGQVQGETLKTGRRFTTS" primer_bind 2484..2500 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 2716..2732 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 2742..2761 /label=T7 /note="T7 promoter, forward primer" promoter 2742..2760 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2808..2828) /label=T3 /note="T3 promoter, forward primer" promoter complement(2808..2826) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2847..2863) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2847..2863) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2860..2882) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2871..2887 /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(2895..2925) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2940..2961 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." ORIGIN 1 acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgacgag acgtctagac 61 cagccaggac agaaatgcct cgacttcgct gctacccaag gttgccgggt gacgcacacc 121 gtggaaacgg atgaaggcac gaacccagtg gacataagcc tgttcggttc gtaagctgta 181 atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg 241 taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg tacagtctat 301 gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag 361 cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca ttatgaggga 421 agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca 481 tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa 541 gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg 601 gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct 661 ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc 721 taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga 781 gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt 841 tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt 901 tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga 961 gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc 1021 gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt 1081 catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc 1141 agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa 1201 ataaccctcg agccacccat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg 1261 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt 1321 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca 1381 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac 1441 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac 1501 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct 1561 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg 1621 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca 1681 gcgtgagcat tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt 1741 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta 1801 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc 1861 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc 1921 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa 1981 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag 2041 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg 2101 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga 2161 gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg 2221 ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct 2281 gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc 2341 ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac 2401 tgagcctttc gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg 2461 gatgttttcc cagtcacgac gttgtaaaac gacggccagt cttaagcgtc tccccctgaa 2521 cctgaccccg gaccaagtgg tggctatcga gaccggcgcc gctacagggc gcgtcccatt 2581 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac 2641 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt 2701 cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac tcactatagg 2761 gcgaattggg taccgggccc cccctcgagg tcctccagct tttgttccct ttagtgaggg 2821 ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg 2881 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa 2941 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca ccggtggtct 3001 c //