LOCUS Exported 4882 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pFA6a-GFP(S65T)-kanMX6 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4882) AUTHORS Bahler J, Wu JQ, Longtine MS, Shah NG, McKenzie A 3rd, Steever AB, Wach A, Philippsen P, Pringle JR TITLE Heterologous modules for efficient and versatile PCR-based gene targeting in Schizosaccharomyces pombe. JOURNAL Yeast. 1998 Jul;14(10):943-51. PUBMED 9717240 REFERENCE 2 (bases 1 to 4882) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4882 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 61..774 /codon_start=1 /product="Aequoria victoria green fluorescent protein" /label=GFP /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF ICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF VTAAGITHGMDELYK" primer_bind complement(88..116) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" primer_bind 711..731 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" terminator 795..982 /gene="S. cerevisiae ADH1" /label=ADH1 terminator /note="transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene" primer_bind complement(850..874) /label=pHybLex-R /note="ADH terminator, reverse primer" gene 1057..2413 /label=kanMX /note="yeast selectable marker conferring kanamycin resistance (Wach et al., 1994)" promoter 1057..1400 /label=TEF promoter /note="Ashbya gossypii TEF promoter" CDS 1401..2210 /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin" /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind complement(1468..1487) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" terminator 2216..2413 /label=TEF terminator /note="Ashbya gossypii TEF terminator" primer_bind complement(2517..2536) /label=T7 /note="T7 promoter, forward primer" promoter complement(2518..2536) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2623..2640) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2794..3382) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2874..2893) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3553..4413) /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 4176..4195 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4414..4518) /gene="bla" /label=AmpR promoter primer_bind 4586..4604 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(4642..4664) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 4764..4783 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 4864..4882 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 4864..4881 /label=SP6 /note="SP6 promoter, forward primer" ORIGIN 1 acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg gttaattaac 61 agtaaaggag aagaactttt cactggagtt gtcccaattc ttgttgaatt agatggtgat 121 gttaatgggc acaaattttc tgtcagtgga gagggtgaag gtgatgcaac atacggaaaa 181 cttaccctta aatttatttg cactactgga aaactacctg ttccatggcc aacacttgtc 241 actactttca cttatggtgt tcaatgcttt tcaagatacc cagatcatat gaaacggcat 301 gactttttca agagtgccat gcccgaaggt tatgtacagg aaagaactat atttttcaaa 361 gatgacggga actacaagac acgtgctgaa gtcaagtttg aaggtgatac ccttgttaat 421 agaatcgagt taaaaggtat tgattttaaa gaagatggaa acattcttgg acacaaattg 481 gaatacaact ataactcaca caatgtatac atcatggcag acaaacaaaa gaatggaatc 541 aaagttaact tcaaaattag acacaacatt gaagatggaa gcgttcaact agcagaccat 601 tatcaacaaa atactccaat tggcgatggc cctgtccttt taccagacaa ccattacctg 661 tccacacaat ctgccctttc gaaagatccc aacgaaaaga gagaccacat ggtccttctt 721 gagtttgtaa cagctgctgg gattacacat ggcatggatg aactatacaa atagggcgcg 781 ccacttctaa ataagcgaat ttcttatgat ttatgatttt tattattaaa taagttataa 841 aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga aaattcttat 901 tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagtat gaggtcgctc 961 ttattgacca cacctctacc ggcagatccg ctagggataa cagggtaata tagatctgtt 1021 tagcttgcct cgtccccgcc gggtcacccg gccagcgaca tggaggccca gaataccctc 1081 cttgacagtc ttgacgtgcg cagctcaggg gcatgatgtg actgtcgccc gtacatttag 1141 cccatacatc cccatgtata atcatttgca tccatacatt ttgatggccg cacggcgcga 1201 agcaaaaatt acggctcctc gctgcagacc tgcgagcagg gaaacgctcc cctcacagac 1261 gcgttgaatt gtccccacgc cgcgcccctg tagagaaata taaaaggtta ggatttgcca 1321 ctgaggttct tctttcatat acttcctttt aaaatcttgc taggatacag ttctcacatc 1381 acatccgaac ataaacaacc atgggtaagg aaaagactca cgtttcgagg ccgcgattaa 1441 attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat 1501 caggtgcgac aatctatcga ttgtatggga agcccgatgc gccagagttg tttctgaaac 1561 atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga 1621 cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt 1681 tactcaccac tgcgatcccc ggcaaaacag cattccaggt attagaagaa tatcctgatt 1741 caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg 1801 tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa 1861 tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg 1921 aacaagtctg gaaagaaatg cataagcttt tgccattctc accggattca gtcgtcactc 1981 atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg 2041 atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc 2101 tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc 2161 ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa tcagtactga 2221 caataaaaag attcttgttt tcaagaactt gtcatttgta tagttttttt atattgtagt 2281 tgttctattt taatcaaatg ttagcgtgat ttatattttt tttcgcctcg acatcatctg 2341 cccagatgcg aagttaagtg cgcagaaagt aatatcatgc gtcaatcgta tgtgaatgct 2401 ggtcgctata ctgctgtcga ttcgatacta acgccgccat ccagtttaaa cgagctcgaa 2461 ttcatcgatg atatcagatc cactagtggc ctatgcggcc gcggatctgc cggtctccct 2521 atagtgagtc gtattaattt cgataagcca ggttaacctg cattaatgaa tcggccaacg 2581 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct 2641 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 2701 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 2761 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 2821 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 2881 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 2941 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg 3001 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 3061 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 3121 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 3181 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt 3241 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 3301 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 3361 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 3421 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 3481 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 3541 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 3601 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 3661 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 3721 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 3781 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 3841 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 3901 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 3961 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 4021 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 4081 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 4141 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 4201 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 4261 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 4321 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 4381 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 4441 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 4501 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat 4561 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg 4621 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg 4681 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg 4741 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat 4801 ggacatattg tcgttagaac gcggctacaa ttaatacata accttatgta tcatacacat 4861 acgatttagg tgacactata ga //