LOCUS       sequence-12697-e        6738 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 6738)
  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 6738)
  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: pXP214
FEATURES             Location/Qualifiers
     source          1..6738
                     /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    complement(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             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    complement(2102..2135)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      2162..3504
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    2494..2541
                     /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(join(3568..3574,3575..3592,3593..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(join(4681..5472,5473..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"
     terminator      complement(5566..5813)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     5779..5797
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     6711..6729
                     /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 cgaattcgta
     2161 tgatccaata tcaaaggaaa tgatagcatt gaaggatgag actaatccaa ttgaggagtg
     2221 gcagcatata gaacagctaa agggtagtgc tgaaggaagc atacgatacc ccgcatggaa
     2281 tgggataata tcacaggagg tactagacta cctttcatcc tacataaata gacgcatata
     2341 agtacgcatt taagcataaa cacgcactat gccgttcttc tcatgtatat atatatacag
     2401 gcaacacgca gatataggtg cgacgtgaac agtgagctgt atgtgcgcag ctcgcgttgc
     2461 attttcggaa gcgctcgttt tcggaaacgc tttgaagttc ctattccgaa gttcctattc
     2521 tctagaaagt ataggaactt cagagcgctt ttgaaaacca aaagcgctct gaagacgcac
     2581 tttcaaaaaa ccaaaaacgc accggactgt aacgagctac taaaatattg cgaataccgc
     2641 ttccacaaac attgctcaaa agtatctctt tgctatatat ctctgtgcta tatccctata
     2701 taacctaccc atccaccttt cgctccttga acttgcatct aaactcgacc tctacatttt
     2761 ttatgtttat ctctagtatt actctttaga caaaaaaatt gtagtaagaa ctattcatag
     2821 agtgaatcga aaacaatacg aaaatgtaaa catttcctat acgtagtata tagagacaaa
     2881 atagaagaaa ccgttcataa ttttctgacc aatgaagaat catcaacgct atcactttct
     2941 gttcacaaag tatgcgcaat ccacatcggt atagaatata atcggggatg cctttatctt
     3001 gaaaaaatgc acccgcagct tcgctagtaa tcagtaaacg cgggaagtgg agtcaggctt
     3061 tttttatgga agagaaaata gacaccaaag tagccttctt ctaaccttaa cggacctaca
     3121 gtgcaaaaag ttatcaagag actgcattat agagcgcaca aaggagaaaa aaagtaatct
     3181 aagatgcttt gttagaaaaa tagcgctctc gggatgcatt tttgtagaac aaaaaagaag
     3241 tatagattct ttgttggtaa aatagcgctc tcgcgttgca tttctgttct gtaaaaatgc
     3301 agctcagatt ctttgtttga aaaattagcg ctctcgcgtt gcatttttgt tttacaaaaa
     3361 tgaagcacag attcttcgtt ggtaaaatag cgctttcgcg ttgcatttct gttctgtaaa
     3421 aatgcagctc agattctttg tttgaaaaat tagcgctctc gcgttgcatt tttgttctac
     3481 aaaatgaagc acagatgctt cgttgaattc 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 tggccgcaaa ttaaagcctt
     5581 cgagcgtccc aaaaccttct caagcaaggt tttcagtata atgttacatg cgtacacgcg
     5641 tctgtacaga aaaaaaagaa aaatttgaaa tataaataac gttcttaata ctaacataac
     5701 tataaaaaaa taaataggga cctagacttc aggttgtcta actccttcct tttcggttag
     5761 agcggatgtg gggggagggc gtgaatgtaa gcgtgacata actaattaca tgactcgagt
     5821 gcgcagatat cactagtata tttgttgtaa aaagtagata attacttcct tgatgatctg
     5881 taaaaaagag aaaaagaaag catctaagaa cttgaaaaac tacgaattag aaaagaccaa
     5941 atatgtattt cttgcattga ccaatttatg caagtttata tatatgtaaa tgtaagtttc
     6001 acgaggttct actaaactaa accaccccct tggttagaag aaaagagtgt gtgagaacag
     6061 gctgttgttg tcacacgatt cggacaattc tgtttgaaag agagagagta acagtacgat
     6121 cgaacgaact ttgctctgga gatcacagtg ggcatcatag catgtggtac taaacccttt
     6181 cccgccattc cagaaccttc gattgcttgt tacaaaacct gtgagccgtc gctaggacct
     6241 tgttgtgtga cgaaattgga agctgcaatc aataggaaga caggaagtcg agcgtgtctg
     6301 ggttttttca gttttgttct ttttgcaaac aaatcacgag cgacggtaat ttctttctcg
     6361 ataagaggcc acgtgcttta tgagggtaac atcaattcaa gaaggaggga aacacttcct
     6421 ttttctggcc ctgataatag tatgagggtg aagccaaaat aaaggattcg cgcccaaatc
     6481 ggcatcttta aatgcaggta tgcgatagtt cctcactctt tccttactca cgagtaattc
     6541 ttgcaaatgc ctaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6601 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6661 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     6721 tcacgaggcc ctttcgtc
//
