LOCUS Exported 5010 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pXP316 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5010) AUTHORS Fang F, Salmon K, Shen MW, Aeling KA, Ito E, Irwin B, Tran UP, Hatfield GW, Da Silva NA, Sandmeyer S TITLE A vector set for systematic metabolic engineering in Saccharomyces cerevisiae. JOURNAL Yeast. 2010 Sep 10. ():. PUBMED 20936606 REFERENCE 2 (bases 1 to 5010) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5010 /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" protein_bind 440..473 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." promoter 474..755 /gene="S. cerevisiae TRP1" /label=TRP1 promoter CDS 756..1430 /codon_start=1 /gene="S. cerevisiae TRP1" /product="phosphoribosylanthranilate isomerase, required for tryptophan biosynthesis" /label=TRP1 /note="yeast auxotrophic marker" /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA KK" protein_bind 1479..1512 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." primer_bind complement(2042..2058) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2042..2058) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2055..2077) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2066..2082 /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(2090..2120) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2135..2156 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2273..2290) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2444..3032) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2524..2543) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3203..4063) /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 3826..3845 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter 4128..4535 /gene="S. cerevisiae TEF1" /label=TEF1 promoter /note="promoter for EF-1-alpha" terminator 4573..4820 /gene="S. cerevisiae CYC1" /label=CYC1 terminator /note="transcription terminator for CYC1" primer_bind complement(4589..4607) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" primer_bind 4983..5001 /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 ccagtgccaa gcttgcatgc ctgcaggtcg 421 actctagagg atccccggga taacttcgta tagcatacat tatacgaagt tataacgaca 481 ttactatata tataatatag gaagcattta atagaacagc atcgtaatat atgtgtactt 541 tgcagttatg acgccagatg gcagtagtgg aagatattct ttattgaaaa atagcttgtc 601 accttacgta caatcttgat ccggagcttt tctttttttg ccgattaaga attaattcgg 661 tcgaaaaaag aaaaggagag ggccaagagg gagggcattg gtgactattg agcacgtgag 721 tatacgtgat taagcacaca aaggcagctt ggagtatgtc tgttattaat ttcacaggta 781 gttctggtcc attggtgaaa gtttgcggct tgcagagcac agaggccgca gaatgtgctc 841 tagattccga tgctgacttg ctgggtatta tatgtgtgcc caatagaaag agaacaattg 901 acccggttat tgcaaggaaa atttcaagtc ttgtaaaagc atataaaaat agttcaggca 961 ctccgaaata cttggttggc gtgtttcgta atcaacctaa ggaggatgtt ttggctctgg 1021 tcaatgatta cggcattgat atcgtccaac tgcatggaga tgagtcgtgg caagaatacc 1081 aagagttcct cggtttgcca gttattaaaa gactcgtatt tccaaaagac tgcaacatac 1141 tactcagtgc agcttcacag aaacctcatt cgtttattcc cttgtttgat tcagaagcag 1201 gtgggacagg tgaacttttg gattggaact cgatttctga ctgggttgga aggcaagaga 1261 gccccgaaag cttacatttt atgttagctg gtggactgac gccagaaaat gttggtgatg 1321 cgcttagatt aaatggcgtt attggtgttg atgtaagcgg aggtgtggag acaaatggtg 1381 taaaagactc taacaaaata gcaaatttcg tcaaaaatgc taagaaatag gttattactg 1441 agtagtattt atttaagtat tgtttgtgca cttgcctgat aacttcgtat agcatacatt 1501 atacgaagtt atcccgggta ccgagctcga attcatcacg tgctataaaa ataattataa 1561 tttaaatttt ttaatataaa tatataaatt aaaaatagaa agtaaaaaaa gaaattaaag 1621 aaaaaatagt ttttgttttc cgaagatgta aaagactcta gggggatcgc caacaaatac 1681 taccttttat cttgctcttc ctgctctcag gtattaatgc cgaattgttt catcttgtct 1741 gtgtagaaga ccacacacga aaatcctgtg attttacatt ttacttatcg ttaatcgaat 1801 gtatatctat ttaatctgct tttcttgtct aataaatata tatgtaaagt acgctttttg 1861 ttgaaatttt ttaaaccttt gtttattttt ttttcttcat tccgtaactc ttctaccttc 1921 tttatttact ttctaaaatc caaatacaaa acataaaaat aaataaacac agagtaaatt 1981 cccaaattat tccatcatta aaagatacga ggcgcgtgta agttacgaat tcgtaatcat 2041 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag 2101 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg 2161 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa 2221 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca 2281 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 2341 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 2401 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 2461 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 2521 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 2581 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata 2641 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 2701 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 2761 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 2821 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 2881 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 2941 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc 3001 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt 3061 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa 3121 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat 3181 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga 3241 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac 3301 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg 3361 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg 3421 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt 3481 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct 3541 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat 3601 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta 3661 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca 3721 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat 3781 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac 3841 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa 3901 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt 3961 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg 4021 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat 4081 attaccgcga atccttacat cacacccaat cccccacaag tgatccccca cacaccatag 4141 cttcaaaatg tttctactcc ttttttactc ttccagattt tctcggactc cgcgcatcgc 4201 cgtaccactt caaaacaccc aagcacagca tactaaattt cccctctttc ttcctctagg 4261 gtgtcgttaa ttacccgtac taaaggtttg gaaaagaaaa aagagaccgc ctcgtttctt 4321 tttcttcgtc gaaaaaggca ataaaaattt ttatcacgtt tctttttctt gaaaattttt 4381 ttttttgatt tttttctctt tcgatgacct cccattgata tttaagttaa taaacggtct 4441 tcaatttctc aagtttcagt ttcatttttc ttgttctatt acaacttttt ttacttcttg 4501 ctcattagaa agaaagcata gcaatctaat ctaagtttta attacaaaac tagtgatatc 4561 tgcgcactcg agtcatgtaa ttagttatgt cacgcttaca ttcacgccct ccccccacat 4621 ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt 4681 ttatagttat gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt 4741 acagacgcgt gtacgcatgt aacattatac tgaaaacctt gcttgagaag gttttgggac 4801 gctcgaaggc tttaatttgc ggccaatatt attgaagcat ttatcagggt tattgtctca 4861 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat 4921 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata 4981 aaaataggcg tatcacgagg ccctttcgtc //