LOCUS Exported 6488 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pXP414 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6488) 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 6488) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6488 /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)." CDS 708..2042 /codon_start=1 /gene="S. cerevisiae MET17" /gene_synonym="MET15; MET25" /product="O-acetylserine and O-acetylhomoserine sulfhydrylase, required for sulfur amino acid synthesis" /label=MET17 /translation="MPSHFDTVQLHAGQENPGDNAHRSRAVPIYATTSYVFENSKHGSQ LFGLEVPGYVYSRFQNPTSNVLEERIAALEGGAAALAVSSGQAAQTLAIQGLAHTGDNI VSTSYLYGGTYNQFKISFKRFGIEARFVEGDNPEEFEKVFDERTKAVYLETIGNPKYNV PDFEKIVAIAHKHGIPVVVDNTFGAGGYFCQPIKYGADIVTHSATKWIGGHGTTIGGII VDSGKFPWKDYPEKFPQFSQPAEGYHGTIYNEAYGNLAYIVHVRTELLRDLGPLMNPFA SFLLLQGVETLSLRAERHGENALKLAKWLEQSPYVSWVSYPGLASHSHHENAKKYLSNG FGGVLSFGVKDLPNADKETDPFKLSGAQVVDNLKLASNLANVGDAKTLVIAPYFTTHKQ LNDKEKLASGVTKDLIRVSVGIEFIDDIIADFQQSFETVFAGQKP" protein_bind 2102..2135 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." rep_origin 2158..3500 /label=2u ori /note="yeast 2u plasmid origin of replication" protein_bind complement(3121..3168) /label=FRT /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." primer_bind complement(3520..3536) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3520..3536) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3533..3555) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3544..3560 /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(3568..3598) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3613..3634 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3751..3768) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3922..4510) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4002..4021) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4681..5541) /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 5304..5323 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter 5606..6013 /gene="S. cerevisiae TEF1" /label=TEF1 promoter /note="promoter for EF-1-alpha" terminator 6051..6298 /gene="S. cerevisiae CYC1" /label=CYC1 terminator /note="transcription terminator for CYC1" primer_bind complement(6067..6085) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" primer_bind 6461..6479 /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 tatgccatcc 481 tcatgaaaac tgtgtaacat aataaccgaa gtgtcgaaaa ggtggcacct tgtccaattg 541 aacacgctcg atgaaaaaaa taagatatat ataaggttaa gtaaagcgtc tgttagaaag 601 gaagtttttc ctttttcttg ctctcttgtc ttttcatcta ctatttcctt cgtgtaatac 661 agggtcgtca gatacataga tacaattcta ttacccccat ccatacaatg ccatctcatt 721 tcgatactgt tcaactacac gccggccaag agaaccctgg tgacaatgct cacagatcca 781 gagctgtacc aatttacgcc accacttctt atgttttcga aaactctaag catggttcgc 841 aattgtttgg tctagaagtt ccaggttacg tctattcccg tttccaaaac ccaaccagta 901 atgttttgga agaaagaatt gctgctttag aaggtggtgc tgctgctttg gctgtttcct 961 ccggtcaagc cgctcaaacc cttgccatcc aaggtttggc acacactggt gacaacatcg 1021 tttccacttc ttacttatac ggtggtactt ataaccagtt caaaatctcg ttcaaaagat 1081 ttggtatcga ggctagattt gttgaaggtg acaatccaga agaattcgaa aaggtctttg 1141 atgaaagaac caaggctgtt tatttggaaa ccattggtaa tccaaagtac aatgttccgg 1201 attttgaaaa aattgttgca attgctcaca aacacggtat tccagttgtc gttgacaaca 1261 catttggtgc cggtggttac ttctgtcagc caattaaata cggtgctgat attgtaacac 1321 attctgctac caaatggatt ggtggtcatg gtactactat cggtggtatt attgttgact 1381 ctggtaagtt cccatggaag gactacccag aaaagttccc tcaattctct caacctgccg 1441 aaggatatca cggtactatc tacaatgaag cctacggtaa cttggcatac atcgttcatg 1501 ttagaactga actattaaga gatttgggtc cattgatgaa cccatttgcc tctttcttgc 1561 tactacaagg tgttgaaaca ttatctttga gagctgaaag acacggtgaa aatgcattga 1621 agttagccaa atggttagaa caatccccat acgtatcttg ggtttcatac cctggtttag 1681 catctcattc tcatcatgaa aatgctaaga agtatctatc taacggtttc ggtggtgtct 1741 tatctttcgg tgtaaaagac ttaccaaatg ccgacaagga aactgaccca ttcaaacttt 1801 ctggtgctca agttgttgac aatttaaagc ttgcctctaa cttggccaat gttggtgatg 1861 ccaagacctt agtcattgct ccatacttca ctacccacaa acaattaaat gacaaagaaa 1921 agttggcatc tggtgttacc aaggacttaa ttcgtgtctc tgttggtatc gaatttattg 1981 atgacattat tgcagacttc cagcaatctt ttgaaactgt tttcgctggc caaaaaccat 2041 gagtgtgcgt aatgagttgt aaaattatgt ataaacctac tttctctcac aagtactata 2101 cataacttcg tatagcatac attatacgaa gttatcccgg gtaccgagct cgaattcaac 2161 gaagcatctg tgcttcattt tgtagaacaa aaatgcaacg cgagagcgct aatttttcaa 2221 acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgaaagc gctattttac 2281 caacgaagaa tctgtgcttc atttttgtaa aacaaaaatg caacgcgaga gcgctaattt 2341 ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg agagcgctat 2401 tttaccaaca aagaatctat acttcttttt tgttctacaa aaatgcatcc cgagagcgct 2461 atttttctaa caaagcatct tagattactt tttttctcct ttgtgcgctc tataatgcag 2521 tctcttgata actttttgca ctgtaggtcc gttaaggtta gaagaaggct actttggtgt 2581 ctattttctc ttccataaaa aaagcctgac tccacttccc gcgtttactg attactagcg 2641 aagctgcggg tgcatttttt caagataaag gcatccccga ttatattcta taccgatgtg 2701 gattgcgcat actttgtgaa cagaaagtga tagcgttgat gattcttcat tggtcagaaa 2761 attatgaacg gtttcttcta ttttgtctct atatactacg tataggaaat gtttacattt 2821 tcgtattgtt ttcgattcac tctatgaata gttcttacta caattttttt gtctaaagag 2881 taatactaga gataaacata aaaaatgtag aggtcgagtt tagatgcaag ttcaaggagc 2941 gaaaggtgga tgggtaggtt atatagggat atagcacaga gatatatagc aaagagatac 3001 ttttgagcaa tgtttgtgga agcggtattc gcaatatttt agtagctcgt tacagtccgg 3061 tgcgtttttg gttttttgaa agtgcgtctt cagagcgctt ttggttttca aaagcgctct 3121 gaagttccta tactttctag agaataggaa cttcggaata ggaacttcaa agcgtttccg 3181 aaaacgagcg cttccgaaaa tgcaacgcga gctgcgcaca tacagctcac tgttcacgtc 3241 gcacctatat ctgcgtgttg cctgtatata tatatacatg agaagaacgg catagtgcgt 3301 gtttatgctt aaatgcgtac ttatatgcgt ctatttatgt aggatgaaag gtagtctagt 3361 acctcctgtg atattatccc attccatgcg gggtatcgta tgcttccttc agcactaccc 3421 tttagctgtt ctatatgctg ccactcctca attggattag tctcatcctt caatgctatc 3481 atttcctttg atattggatc atacgaattc gtaatcatgg tcatagctgt ttcctgtgtg 3541 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc 3601 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt 3661 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg 3721 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt 3781 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 3841 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 3901 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 3961 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 4021 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 4081 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 4141 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 4201 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 4261 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 4321 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg 4381 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 4441 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 4501 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 4561 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 4621 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 4681 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 4741 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 4801 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 4861 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 4921 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 4981 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 5041 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 5101 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 5161 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 5221 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 5281 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 5341 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 5401 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 5461 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 5521 acggaaatgt tgaatactca tactcttcct ttttcaatat taccgcgaat ccttacatca 5581 cacccaatcc cccacaagtg atcccccaca caccatagct tcaaaatgtt tctactcctt 5641 ttttactctt ccagattttc tcggactccg cgcatcgccg taccacttca aaacacccaa 5701 gcacagcata ctaaatttcc cctctttctt cctctagggt gtcgttaatt acccgtacta 5761 aaggtttgga aaagaaaaaa gagaccgcct cgtttctttt tcttcgtcga aaaaggcaat 5821 aaaaattttt atcacgtttc tttttcttga aaattttttt ttttgatttt tttctctttc 5881 gatgacctcc cattgatatt taagttaata aacggtcttc aatttctcaa gtttcagttt 5941 catttttctt gttctattac aacttttttt acttcttgct cattagaaag aaagcatagc 6001 aatctaatct aagttttaat tacaaaacta gtgatatctg cgcactcgag tcatgtaatt 6061 agttatgtca cgcttacatt cacgccctcc ccccacatcc gctctaaccg aaaaggaagg 6121 agttagacaa cctgaagtct aggtccctat ttattttttt atagttatgt tagtattaag 6181 aacgttattt atatttcaaa tttttctttt ttttctgtac agacgcgtgt acgcatgtaa 6241 cattatactg aaaaccttgc ttgagaaggt tttgggacgc tcgaaggctt taatttgcgg 6301 ccaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg 6361 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga 6421 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc 6481 ctttcgtc //