LOCUS Exported 4161 bp ds-DNA linear SYN 20-MAY-2021 DEFINITION Expression of photoactivated cyclase bPAC of Beggiatoa sp.. ACCESSION . VERSION . KEYWORDS bPAC-mycHis_pGEM SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4161) AUTHORS Stierl M, Stumpf P, Udwari D, Gueta R, Hagedorn R, Losi A, Gartner W, Petereit L, Efetova M, Schwarzel M, Oertner TG, Nagel G, Hegemann P TITLE Light modulation of cellular cAMP by a small bacterial photoactivated adenylyl cyclase, bPAC, of the soil bacterium Beggiatoa. JOURNAL J Biol Chem. 2011 Jan 14. 286(2):1181-8. PUBMED 21030594 REFERENCE 2 (bases 1 to 4161) AUTHORS . TITLE Direct Submission JOURNAL Exported May 20, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4161 /organism="synthetic DNA construct" /mol_type="other DNA" promoter complement(70..88) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(71..88) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(106..122) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(106..122) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(119..141) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 130..146 /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(154..184) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 199..220 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(337..354) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(508..1096) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(588..607) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1267..2127) /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 1890..1909 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2128..2232) /gene="bla" /label=AmpR promoter primer_bind 2300..2318 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(2356..2378) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 2478..2497 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 2691..2713 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 2705..2722 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 2706..2722 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 2729..2748 /label=T7 /note="T7 promoter, forward primer" promoter 2729..2747 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" 5'UTR 2779..2821 /label=Xenopus globin 5'-UTR /note="translational enhancer from the 5'-UTR of the major beta-globin gene of Xenopus laevis" CDS 3918..3947 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 3948..3977 /codon_start=1 /product="10xHis affinity tag" /label=10xHis /translation="HHHHHHHHHH" 3'UTR 4026..4151 /label=Xenopus globin 3'-UTR /note="3'-UTR of the major beta-globin gene of Xenopus laevis" ORIGIN 1 cccccccctg caggcatgcg tttaaacatt taaattacgt attaattaag atatcgctag 61 cttgagtatt ctatagtgtc acctaaatag cttggcgtaa tcatggtcat agctgtttcc 121 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 181 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 241 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 301 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 361 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 421 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 481 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 541 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 601 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 661 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 721 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 781 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 841 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 901 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg 961 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 1021 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 1081 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 1141 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 1201 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 1261 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 1321 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 1381 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 1441 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 1501 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 1561 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 1621 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 1681 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 1741 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 1801 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 1861 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 1921 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 1981 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 2041 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 2101 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 2161 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 2221 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat 2281 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg 2341 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta 2401 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg 2461 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg 2521 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag 2581 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc 2641 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg 2701 acgttgtaaa acgacggcca gtgaattgta atacgactca ctatagggcg aattaattcg 2761 agctcggtac ccagcttgct tgttcttttt gcagaagctc agaataaacg ctcaactttg 2821 gcagatcaat tccccgggga tccaagcttg ccaccatgat gaagcggctg gtgtacatca 2881 gcaagatcag cggccacctg agcctggaag agatccagcg gatcggcaag gtgtccatca 2941 agaacaacca gcgggacaac atcaccggcg tgctgctgta cctgcagggc ctgttcttcc 3001 agatcctgga aggcgagaac gagaaggtgg acaagctgta caagaaaatc ctggtggacg 3061 accggcacac caacatcctg tgcctgaaaa ccgagtacga catcaccgac cggatgttcc 3121 ccaactgggc catgaaaacc atcaacctga acgagaacag cgagctgatg atccagccca 3181 tcaagagcct gctgcagacc atcacccaga gccaccgggt gctggaaaag tacatgcccg 3241 ccagagtgat ctacctgatc aaccagggca tcaaccccct gaccgtggag ccccagctgg 3301 tcgagaagat catcttcttc agcgacatcc tggccttcag caccctgacc gagaagctgc 3361 ccgtgaacga ggtggtcatc ctggtcaacc ggtacttcag catctgcacc cggatcatca 3421 gcgcctacgg cggcgaagtg accaagttca tcggcgactg cgtgatggcc agcttcacca 3481 aagagcaggg cgacgccgcc atccggacca gcctggacat catctccgag ctgaagcagc 3541 tgcggcacca cgtggaggcc accaaccccc tgcacctgct gtacaccggc atcggcctga 3601 gctacggcca cgtgatcgag ggcaacatgg gcagcagcct gaagatggac cacaccctgc 3661 tgggggacgc cgtgaacgtg gccgccaggc tggaagccct gacaagacag ctgccctacg 3721 ccctggcctt tacagccggc gtgaagaagt gctgccaggc ccagtggacc ttcatcaacc 3781 tgggagccca ccaggtcaag ggcaagcagg aagccatcga ggtctacacc gtgaatgagg 3841 cccagaagta ctacgacacc ctgcagatca cccagctgat ccggcagacc ctggaaaacg 3901 acaagccgcg gacgtacgag caaaagctga tctcggagga ggacctgcac catcaccatc 3961 accatcacca tcaccatctc gagtaagcgg ccgcaggacc tgtaagcttg atctggttac 4021 cactaaacca gcctcaagaa cacccgaatg gagtctctaa gctacataat accaacttac 4081 actttacaaa atgttgtccc ccaaaatgta gccattcgta tctgctccta ataaaaagaa 4141 agtttcttca cattctaaaa a //