LOCUS Exported 4078 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pFA6a-5FLAG-natMX6 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4078) AUTHORS Noguchi C, Garabedian MV, Malik M, Noguchi E TITLE A vector system for genomic FLAG epitope-tagging in Schizosaccharomyces pombe. JOURNAL Biotechnol J. 2008 Aug 26. ():. PUBMED 18729046 REFERENCE 2 (bases 1 to 4078) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4078 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 69..200 /codon_start=1 /product="five tandem FLAG(R) epitope tags" /label=5xFLAG /translation="DYKDDDDKDYKDDDDKLMDYKDDDDKDYKDDDDKLMDYKDDDDK" terminator 227..414 /gene="S. cerevisiae ADH1" /label=ADH1 terminator /note="transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene" primer_bind complement(282..306) /label=pHybLex-R /note="ADH terminator, reverse primer" gene 489..1611 /label=natMX6 /note="yeast selectable marker conferring nourseothricin resistance" promoter 489..832 /label=TEF promoter /note="Ashbya gossypii TEF promoter" CDS 833..1408 /codon_start=1 /gene="Streptomyces noursei nat1" /product="nourseothricin acetyltransferase" /label=NrsR /note="confers resistance to nourseothricin" /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVISYSAWNR RLTVEDIEVAPEHRGHGVGRALMGLATEFAGERGAGHLWLEVTNVNAPAIHAYRRMGFT LCGLDTALYDGTASDGERQALYMSMPCP" terminator 1414..1611 /label=TEF terminator /note="Ashbya gossypii TEF terminator" primer_bind complement(1715..1734) /label=T7 /note="T7 promoter, forward primer" promoter complement(1716..1734) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1821..1838) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1992..2580) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2072..2091) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2751..3611) /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 3374..3393 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3612..3716) /gene="bla" /label=AmpR promoter primer_bind 3784..3802 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(3840..3862) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 3962..3981 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 4062..2 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 4062..1 /label=SP6 /note="SP6 promoter, forward primer" ORIGIN 1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta 61 atcatatgga ctacaaggac gacgatgaca aggattacaa agatgacgac gataagctta 121 tggactacaa ggacgacgat gacaaggatt acaaagatga cgacgataag cttatggact 181 acaaggacga cgatgacaag catatgggcg cgccacttct aaataagcga atttcttatg 241 atttatgatt tttattatta aataagttat aaaaaaaata agtgtataca aattttaaag 301 tgactcttag gttttaaaac gaaaattctt attcttgagt aactctttcc tgtaggtcag 361 gttgctttct caggtatagt atgaggtcgc tcttattgac cacacctcta ccggcagatc 421 cgctagggat aacagggtaa tatagatctg tttagcttgc ctcgtccccg ccgggtcacc 481 cggccagcga catggaggcc cagaataccc tccttgacag tcttgacgtg cgcagctcag 541 gggcatgatg tgactgtcgc ccgtacattt agcccataca tccccatgta taatcatttg 601 catccataca ttttgatggc cgcacggcgc gaagcaaaaa ttacggctcc tcgctgcaga 661 cctgcgagca gggaaacgct cccctcacag acgcgttgaa ttgtccccac gccgcgcccc 721 tgtagagaaa tataaaaggt taggatttgc cactgaggtt cttctttcat atacttcctt 781 ttaaaatctt gctaggatac agttctcaca tcacatccga acataaacaa ccatgggtac 841 cactcttgac gacacggctt accggtaccg caccagtgtc ccgggggacg ccgaggccat 901 cgaggcactg gatgggtcct tcaccaccga caccgtcttc cgcgtcaccg ccaccgggga 961 cggcttcacc ctgcgggagg tgccggtgga cccgcccctg accaaggtgt tccccgacga 1021 cgaatcggac gacgaatcgg acgacgggga ggacggcgac ccggactccc ggacgttcgt 1081 cgcgtacggg gacgacggcg acctggcggg cttcgtggtc atctcgtact cggcgtggaa 1141 ccgccggctg accgtcgagg acatcgaggt cgccccggag caccgggggc acggggtcgg 1201 gcgcgcgttg atggggctcg cgacggagtt cgccggcgag cggggcgccg ggcacctctg 1261 gctggaggtc accaacgtca acgcaccggc gatccacgcg taccggcgga tggggttcac 1321 cctctgcggc ctggacaccg ccctgtacga cggcaccgcc tcggacggcg agcggcaggc 1381 gctctacatg agcatgccct gcccctaatc agtactgaca ataaaaagat tcttgttttc 1441 aagaacttgt catttgtata gtttttttat attgtagttg ttctatttta atcaaatgtt 1501 agcgtgattt atattttttt tcgcctcgac atcatctgcc cagatgcgaa gttaagtgcg 1561 cagaaagtaa tatcatgcgt caatcgtatg tgaatgctgg tcgctatact gctgtcgatt 1621 cgatactaac gccgccatcc agtttaaacg agctcgaatt catcgatgat atcagatcca 1681 ctagtggcct atgcggccgc ggatctgccg gtctccctat agtgagtcgt attaatttcg 1741 ataagccagg ttaacctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt 1801 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg 1861 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac 1921 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg 1981 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca 2041 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc 2101 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc 2161 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag 2221 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc 2281 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca 2341 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg 2401 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg 2461 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct 2521 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa 2581 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa 2641 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa 2701 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc 2761 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga 2821 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca 2881 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc 2941 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat 3001 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc 3061 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt 3121 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc 3181 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg 3241 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt 3301 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg 3361 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga 3421 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg 3481 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg 3541 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt 3601 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc 3661 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca 3721 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat 3781 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac 3841 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc 3901 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat 3961 gcggcatcag agcagattgt actgagagtg caccatatgg acatattgtc gttagaacgc 4021 ggctacaatt aatacataac cttatgtatc atacacatac gatttaggtg acactata //