LOCUS       sequence-140301-        5882 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 5882)
  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 5882)
  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: pXP114
FEATURES             Location/Qualifiers
     source          1..5882
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    750..783
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(843..2177)
                     /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    2412..2445
                     /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(2490..2507)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2490..2506)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2499..2521)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2715..2734)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2834..2856
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2894..2912)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      3810..4057
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3826..3844)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             join(4082..4150,4151..4942)
                     /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     complement(4300..4319)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5113..5701
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5602..5621
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5855..5872
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga catgattacg aattcatcac
      241 gtgctataaa aataattata atttaaattt tttaatataa atatataaat taaaaataga
      301 aagtaaaaaa agaaattaaa gaaaaaatag tttttgtttt ccgaagatgt aaaagactct
      361 agggggatcg ccaacaaata ctacctttta tcttgctctt cctgctctca ggtattaatg
      421 ccgaattgtt tcatcttgtc tgtgtagaag accacacacg aaaatcctgt gattttacat
      481 tttacttatc gttaatcgaa tgtatatcta tttaatctgc ttttcttgtc taataaatat
      541 atatgtaaag tacgcttttt gttgaaattt tttaaacctt tgtttatttt tttttcttca
      601 ttccgtaact cttctacctt ctttatttac tttctaaaat ccaaatacaa aacataaaaa
      661 taaataaaca cagagtaaat tcccaaatta ttccatcatt aaaagatacg aggcgcgtgt
      721 aagttacgaa ttcgagctcg gtacccggga taacttcgta taatgtatgc tatacgaagt
      781 tatgtatagt acttgtgaga gaaagtaggt ttatacataa ttttacaact cattacgcac
      841 actcatggtt tttggccagc gaaaacagtt tcaaaagatt gctggaagtc tgcaataatg
      901 tcatcaataa attcgatacc aacagagaca cgaattaagt ccttggtaac accagatgcc
      961 aacttttctt tgtcatttaa ttgtttgtgg gtagtgaagt atggagcaat gactaaggtc
     1021 ttggcatcac caacattggc caagttagag gcaagcttta aattgtcaac aacttgagca
     1081 ccagaaagtt tgaatgggtc agtttccttg tcggcatttg gtaagtcttt tacaccgaaa
     1141 gataagacac caccgaaacc gttagataga tacttcttag cattttcatg atgagaatga
     1201 gatgctaaac cagggtatga aacccaagat acgtatgggg attgttctaa ccatttggct
     1261 aacttcaatg cattttcacc gtgtctttca gctctcaaag ataatgtttc aacaccttgt
     1321 agtagcaaga aagaggcaaa tgggttcatc aatggaccca aatctcttaa tagttcagtt
     1381 ctaacatgaa cgatgtatgc caagttaccg taggcttcat tgtagatagt accgtgatat
     1441 ccttcggcag gttgagagaa ttgagggaac ttttctgggt agtccttcca tgggaactta
     1501 ccagagtcaa caataatacc accgatagta gtaccatgac caccaatcca tttggtagca
     1561 gaatgtgtta caatatcagc accgtattta attggctgac agaagtaacc accggcacca
     1621 aatgtgttgt caacgacaac tggaataccg tgtttgtgag caattgcaac aattttttca
     1681 aaatccggaa cattgtactt tggattacca atggtttcca aataaacagc cttggttctt
     1741 tcatcaaaga ccttttcgaa ttcttctgga ttgtcacctt caacaaatct agcctcgata
     1801 ccaaatcttt tgaacgagat tttgaactgg ttataagtac caccgtataa gtaagaagtg
     1861 gaaacgatgt tgtcaccagt gtgtgccaaa ccttggatgg caagggtttg agcggcttga
     1921 ccggaggaaa cagccaaagc agcagcacca ccttctaaag cagcaattct ttcttccaaa
     1981 acattactgg ttgggttttg gaaacgggaa tagacgtaac ctggaacttc tagaccaaac
     2041 aattgcgaac catgcttaga gttttcgaaa acataagaag tggtggcgta aattggtaca
     2101 gctctggatc tgtgagcatt gtcaccaggg ttctcttggc cggcgtgtag ttgaacagta
     2161 tcgaaatgag atggcattgt atggatgggg gtaatagaat tgtatctatg tatctgacga
     2221 ccctgtatta cacgaaggaa atagtagatg aaaagacaag agagcaagaa aaaggaaaaa
     2281 cttcctttct aacagacgct ttacttaacc ttatatatat cttatttttt tcatcgagcg
     2341 tgttcaattg gacaaggtgc caccttttcg acacttcggt tattatgtta cacagttttc
     2401 atgaggatgg cataacttcg tataatgtat gctatacgaa gttatcccgg ggatcctcta
     2461 gagtcgacct gcaggcatgc aagcttggca ctggccgtcg ttttacaacg tcgtgactgg
     2521 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     2581 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     2641 gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     2701 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     2761 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     2821 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     2881 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     2941 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     3001 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     3061 caataatatt aggcatttgc aagaattact cgtgagtaag gaaagagtga ggaactatcg
     3121 catacctgca tttaaagatg ccgatttggg cgcgaatcct ttattttggc ttcaccctca
     3181 tactattatc agggccagaa aaaggaagtg tttccctcct tcttgaattg atgttaccct
     3241 cataaagcac gtggcctctt atcgagaaag aaattaccgt cgctcgtgat ttgtttgcaa
     3301 aaagaacaaa actgaaaaaa cccagacacg ctcgacttcc tgtcttccta ttgattgcag
     3361 cttccaattt cgtcacacaa caaggtccta gcgacggctc acaggttttg taacaagcaa
     3421 tcgaaggttc tggaatggcg ggaaagggtt tagtaccaca tgctatgatg cccactgtga
     3481 tctccagagc aaagttcgtt cgatcgtact gttactctct ctctttcaaa cagaattgtc
     3541 cgaatcgtgt gacaacaaca gcctgttctc acacactctt ttcttctaac caagggggtg
     3601 gtttagttta gtagaacctc gtgaaactta catttacata tatataaact tgcataaatt
     3661 ggtcaatgca agaaatacat atttggtctt ttctaattcg tagtttttca agttcttaga
     3721 tgctttcttt ttctcttttt tacagatcat caaggaagta attatctact ttttacaaca
     3781 aatatactag tgatatctgc gcactcgagt catgtaatta gttatgtcac gcttacattc
     3841 acgccctccc cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta
     3901 ggtccctatt tattttttta tagttatgtt agtattaaga acgttattta tatttcaaat
     3961 ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac attatactga aaaccttgct
     4021 tgagaaggtt ttgggacgct cgaaggcttt aatttgcggc caatattgaa aaaggaagag
     4081 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     4141 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     4201 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     4261 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     4321 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     4381 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     4441 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     4501 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     4561 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     4621 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     4681 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     4741 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     4801 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     4861 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     4921 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     4981 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     5041 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     5101 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5161 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     5221 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     5281 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     5341 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     5401 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     5461 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     5521 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     5581 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     5641 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     5701 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     5761 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     5821 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     5881 ag
//
