LOCUS       sequence-12715-e        5150 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 5150)
  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 5150)
  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: pXP522
FEATURES             Location/Qualifiers
     source          1..5150
                     /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"
     terminator      598..845
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(614..632)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     protein_bind    complement(891..924)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(2366..2399)
                     /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(2929..2945)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2929..2945)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2942..2964)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2953..2969
                     /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(2977..2983,2984..3001,3002..3007))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3022..3043
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3160..3177)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3331..3919)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3411..3430)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4090..4881,4882..4950))
                     /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     4713..4732
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4951..5055)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5123..5141
                     /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 accatatgct cgtaggaaca atttcgggcc cctgcgtgtt cttctgaggt tcatctttta
      241 catttgcttc tgctggataa ttttcagagg caacaaggaa aaattagatg gcaaaaagtc
      301 gtctttcaag gaaaaatccc caccatcttt cgagatcccc tgtaacttat tggcaactga
      361 aagaatgaaa aggaggaaaa tacaaaatat actagaactg aaaaaaaaaa agtataaata
      421 gagacgatat atgccaatac ttcacaatgt tcgaatctat tcttcatttg cagctattgt
      481 aaaataataa aacatcaaga acaaacaagc tcaacttgtc ttttctaaga acaaagaata
      541 aacacaaaaa caaaaagttt ttttaatttt aatcaaaaaa ctagtgatat cctcgagtca
      601 tgtaattagt tatgtcacgc ttacattcac gccctccccc cacatccgct ctaaccgaaa
      661 aggaaggagt tagacaacct gaagtctagg tccctattta tttttttata gttatgttag
      721 tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg
      781 catgtaacat tatactgaaa accttgcttg agaaggtttt gggacgctcg aaggctttaa
      841 tttgcggcca agcttgcatg cctgcaggtc gactctagag gatccccggg ataacttcgt
      901 atagcataca ttatacgaag ttattaacca ttattttttt cctcaacata acgagaacac
      961 acaggggcgc tatcgcacag aatcaaattc gatgactgga aattttttgt taatttcaga
     1021 ggtcgcctga cgcatatacc tttttcaact gaaaaattgg gagaaaaagg aaaggtgaga
     1081 ggccggaacc ggcttttcat atagaataga gaagcgttca tgactaaatg cttgcatcac
     1141 aatacttgaa gttgacaata ttatttaagg acctattgtt ttttccaata ggtggttagc
     1201 aatcgtctta ctttctaact tttcttacct tttacatttc agcaatatat atatatattt
     1261 caaggatata ccattctaat gtctgcccct aagaagatcg tcgttttgcc aggtgaccac
     1321 gttggtcaag aaatcacagc cgaagccatt aaggttctta aagctatttc tgatgttcgt
     1381 tccaatgtca agttcgattt cgaaaatcat ttaattggtg gtgctgctat cgaagctaca
     1441 ggtgtcccac ttccagatga ggcgctggaa gcctccaaga aggctgatgc cgttttgtta
     1501 ggtgctgtgg gtggtcctaa atggggtaca ggtagtgtta gacctgaaca aggtttacta
     1561 aaaatccgta aagaacttca attgtacgcc aacttaagac catgtaactt tgcatccgac
     1621 tctcttttag acttatctcc aatcaagcca caatttgcta aaggtactga cttcgttgtt
     1681 gtcagagaat tagtgggagg tatttacttt ggtaagagaa aggaagatga tggtgatggt
     1741 gtcgcttggg atagtgaaca atacaccgtt ccagaagtgc aaagaatcac aagaatggcc
     1801 gctttcatgg ccctacaaca tgagccacca ttgcctattt ggtccttgga taaagctaat
     1861 gttttggcct cttcaagatt atggagaaaa actgtggagg aaaccatcaa gaacgaattt
     1921 cctacattga aggttcaaca tcaattgatt gattctgccg ccatgatcct agttaagaac
     1981 ccaacccacc taaatggtat tataatcacc agcaacatgt ttggtgatat catctccgat
     2041 gaagcctccg ttatcccagg ttccttgggt ttgttgccat ctgcgtcctt ggcctctttg
     2101 ccagacaaga acaccgcatt tggtttgtac gaaccatgcc acggttctgc tccagatttg
     2161 ccaaagaata aggtcaaccc tatcgccact atcttgtctg ctgcaatgat gttgaaattg
     2221 tcattgaact tgcctgaaga aggtaaggcc attgaagatg cagttaaaaa ggttttggat
     2281 gcaggtatca gaactggtga tttaggtggt tccaacagta ccaccgaagt cggtgatgct
     2341 gtcgccgaag aagttaagaa aatccataac ttcgtatagc atacattata cgaagttatc
     2401 ccgggtaccg agctcgaatt cgtaacttac acgcgcctcg tatcttttaa tgatggaata
     2461 atttgggaat ttactctgtg tttatttatt tttatgtttt gtatttggat tttagaaagt
     2521 aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa
     2581 atttcaacaa aaagcgtact ttacatatat atttattaga caagaaaagc agattaaata
     2641 gatatacatt cgattaacga taagtaaaat gtaaaatcac aggattttcg tgtgtggtct
     2701 tctacacaga caagatgaaa caattcggca ttaatacctg agagcaggaa gagcaagata
     2761 aaaggtagta tttgttggcg atccccctag agtcttttac atcttcggaa aacaaaaact
     2821 attttttctt taatttcttt ttttactttc tatttttaat ttatatattt atattaaaaa
     2881 atttaaatta taattatttt tatagcacgt gatgaattcg taatcatggt catagctgtt
     2941 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     3001 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     3061 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     3121 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     3181 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3241 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3301 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3361 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3421 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3481 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3541 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3601 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3661 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3721 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     3781 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3841 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3901 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3961 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4021 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4081 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4141 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4201 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4261 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4321 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4381 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4441 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4501 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4561 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4621 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4681 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4741 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4801 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4861 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4921 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4981 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5041 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     5101 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
//
