LOCUS       sequence-12703-e        5633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5633)
  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. 2011 Feb;28(2):123-36. doi: 10.1002/yea.1824. Epub 2010 Oct 
            8.
  PUBMED    20936606
REFERENCE   2  (bases 1 to 5633)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1002/yea.1824"; 
            journalName: "Yeast"; date: "2011-02"; volume: "28"; issue: "2"; 
            pages: "123-36"
COMMENT     Sequence Label: pXP314
FEATURES             Location/Qualifiers
     source          1..5633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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 (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(533..1867)
                     /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 (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(2665..2681)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2665..2681)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2678..2700)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2689..2705
                     /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(join(2713..2719,2720..2737,2738..2743))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2758..2779
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2896..2913)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3067..3655)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3147..3166)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3826..4617,4618..4686))
                     /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     4449..4468
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        4745..5158
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      5196..5443
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5212..5230)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     5606..5624
                     /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 taatgtatgc tatacgaagt tatgtatagt
      481 acttgtgaga gaaagtaggt ttatacataa ttttacaact cattacgcac actcatggtt
      541 tttggccagc gaaaacagtt tcaaaagatt gctggaagtc tgcaataatg tcatcaataa
      601 attcgatacc aacagagaca cgaattaagt ccttggtaac accagatgcc aacttttctt
      661 tgtcatttaa ttgtttgtgg gtagtgaagt atggagcaat gactaaggtc ttggcatcac
      721 caacattggc caagttagag gcaagcttta aattgtcaac aacttgagca ccagaaagtt
      781 tgaatgggtc agtttccttg tcggcatttg gtaagtcttt tacaccgaaa gataagacac
      841 caccgaaacc gttagataga tacttcttag cattttcatg atgagaatga gatgctaaac
      901 cagggtatga aacccaagat acgtatgggg attgttctaa ccatttggct aacttcaatg
      961 cattttcacc gtgtctttca gctctcaaag ataatgtttc aacaccttgt agtagcaaga
     1021 aagaggcaaa tgggttcatc aatggaccca aatctcttaa tagttcagtt ctaacatgaa
     1081 cgatgtatgc caagttaccg taggcttcat tgtagatagt accgtgatat ccttcggcag
     1141 gttgagagaa ttgagggaac ttttctgggt agtccttcca tgggaactta ccagagtcaa
     1201 caataatacc accgatagta gtaccatgac caccaatcca tttggtagca gaatgtgtta
     1261 caatatcagc accgtattta attggctgac agaagtaacc accggcacca aatgtgttgt
     1321 caacgacaac tggaataccg tgtttgtgag caattgcaac aattttttca aaatccggaa
     1381 cattgtactt tggattacca atggtttcca aataaacagc cttggttctt tcatcaaaga
     1441 ccttttcgaa ttcttctgga ttgtcacctt caacaaatct agcctcgata ccaaatcttt
     1501 tgaacgagat tttgaactgg ttataagtac caccgtataa gtaagaagtg gaaacgatgt
     1561 tgtcaccagt gtgtgccaaa ccttggatgg caagggtttg agcggcttga ccggaggaaa
     1621 cagccaaagc agcagcacca ccttctaaag cagcaattct ttcttccaaa acattactgg
     1681 ttgggttttg gaaacgggaa tagacgtaac ctggaacttc tagaccaaac aattgcgaac
     1741 catgcttaga gttttcgaaa acataagaag tggtggcgta aattggtaca gctctggatc
     1801 tgtgagcatt gtcaccaggg ttctcttggc cggcgtgtag ttgaacagta tcgaaatgag
     1861 atggcattgt atggatgggg gtaatagaat tgtatctatg tatctgacga ccctgtatta
     1921 cacgaaggaa atagtagatg aaaagacaag agagcaagaa aaaggaaaaa cttcctttct
     1981 aacagacgct ttacttaacc ttatatatat cttatttttt tcatcgagcg tgttcaattg
     2041 gacaaggtgc caccttttcg acacttcggt tattatgtta cacagttttc atgaggatgg
     2101 cataacttcg tataatgtat gctatacgaa gttatcccgg gtaccgagct cgaattcatc
     2161 acgtgctata aaaataatta taatttaaat tttttaatat aaatatataa attaaaaata
     2221 gaaagtaaaa aaagaaatta aagaaaaaat agtttttgtt ttccgaagat gtaaaagact
     2281 ctagggggat cgccaacaaa tactaccttt tatcttgctc ttcctgctct caggtattaa
     2341 tgccgaattg tttcatcttg tctgtgtaga agaccacaca cgaaaatcct gtgattttac
     2401 attttactta tcgttaatcg aatgtatatc tatttaatct gcttttcttg tctaataaat
     2461 atatatgtaa agtacgcttt ttgttgaaat tttttaaacc tttgtttatt tttttttctt
     2521 cattccgtaa ctcttctacc ttctttattt actttctaaa atccaaatac aaaacataaa
     2581 aataaataaa cacagagtaa attcccaaat tattccatca ttaaaagata cgaggcgcgt
     2641 gtaagttacg aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     2701 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     2761 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     2821 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     2881 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     2941 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3001 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3061 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3121 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3181 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3241 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3301 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3361 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3421 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3481 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     3541 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3601 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3661 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3721 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3781 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     3841 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3901 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     3961 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4021 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4081 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4141 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4201 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4261 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4321 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4381 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4441 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4501 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4561 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4621 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4681 actcatactc ttcctttttc aatattaccg cgaatcctta catcacaccc aatcccccac
     4741 aagtgatccc ccacacacca tagcttcaaa atgtttctac tcctttttta ctcttccaga
     4801 ttttctcgga ctccgcgcat cgccgtacca cttcaaaaca cccaagcaca gcatactaaa
     4861 tttcccctct ttcttcctct agggtgtcgt taattacccg tactaaaggt ttggaaaaga
     4921 aaaaagagac cgcctcgttt ctttttcttc gtcgaaaaag gcaataaaaa tttttatcac
     4981 gtttcttttt cttgaaaatt tttttttttg atttttttct ctttcgatga cctcccattg
     5041 atatttaagt taataaacgg tcttcaattt ctcaagtttc agtttcattt ttcttgttct
     5101 attacaactt tttttacttc ttgctcatta gaaagaaagc atagcaatct aatctaagtt
     5161 ttaattacaa aactagtgat atctgcgcac tcgagtcatg taattagtta tgtcacgctt
     5221 acattcacgc cctcccccca catccgctct aaccgaaaag gaaggagtta gacaacctga
     5281 agtctaggtc cctatttatt tttttatagt tatgttagta ttaagaacgt tatttatatt
     5341 tcaaattttt cttttttttc tgtacagacg cgtgtacgca tgtaacatta tactgaaaac
     5401 cttgcttgag aaggttttgg gacgctcgaa ggctttaatt tgcggccaat attattgaag
     5461 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5521 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     5581 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtc
//
