LOCUS       sequence-64880-e        5336 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 5336)
  AUTHORS   Shen MW, Fang F, Sandmeyer S, Da Silva NA
  TITLE     Development and characterization of a vector set with regulated 
            promoters for systematic metabolic engineering in Saccharomyces 
            cerevisiae.
  JOURNAL   Yeast. 2012 Dec;29(12):495-503. doi: 10.1002/yea.2930. Epub 2012 Nov
            19.
  PUBMED    23166051
REFERENCE   2  (bases 1 to 5336)
  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.2930"; 
            journalName: "Yeast"; date: "2012-12"; volume: "29"; issue: "12"; 
            pages: "495-503"
COMMENT     Sequence Label: pXP741
FEATURES             Location/Qualifiers
     source          1..5336
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      602..849
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(618..636)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     protein_bind    895..928
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2370..2403
                     /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(2933..2949)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2933..2949)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2946..2968)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2957..2973
                     /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(2981..2987,2988..3005,3006..3011))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3026..3047
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3164..3181)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3335..3923)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3415..3434)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4094..4885,4886..4954))
                     /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     4717..4736
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4955..5059)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5127..5145
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5183..5205)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5305..5324
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 catatgattt agcggccgca aaacgtaggg gcaaacaaac ggaaaaatcg tttctcaaat
       61 tttctgatgc caagaactct aaccagtctt atctaaaaat tgccttatga tccgtctctc
      121 cggttacagc ctgtgtaact gattaatcct gcctttctaa tcaccattct aatgttttaa
      181 ttaagggatt ttgtcttcat taacggcttt cgctcataaa aatgttatga cgttttgccc
      241 gcaggcggga aaccatccac ttcacgagac tgatctcctc tgccggaaca ccgggcatct
      301 ccaacttata agttggagaa ataagagaat ttcagattga gagaatgaaa aaaaaaaaaa
      361 aaaaaaaggc agaggagagc atagaaatgg ggttcacttt ttggtaaagc tatagcatgc
      421 ctatcacata taaatagagt gccagtagcg acttttttca cactcgaaat actcttacta
      481 ctgctctctt gttgttttta tcacttcttg tttcttcttg gtaaatagaa tatcaagcta
      541 caaaaagcat acaatcaact atcaactatt aactatatcg taatacacaa ctagtctcga
      601 gtcatgtaat tagttatgtc acgcttacat tcacgccctc cccccacatc cgctctaacc
      661 gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt tatagttatg
      721 ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta cagacgcgtg
      781 tacgcatgta acattatact gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct
      841 ttaatttgcg gccaagcttg catgcctgca ggtcgactct agaggatccc cgggataact
      901 tcgtataatg tatgctatac gaagttatgg attttcttaa cttcttcggc gacagcatca
      961 ccgacttcgg tggtactgtt ggaaccacct aaatcaccag ttctgatacc tgcatccaaa
     1021 acctttttaa ctgcatcttc aatggcctta ccttcttcag gcaagttcaa tgacaatttc
     1081 aacatcattg cagcagacaa gatagtggcg atagggttga ccttattctt tggcaaatct
     1141 ggagcagaac cgtggcatgg ttcgtacaaa ccaaatgcgg tgttcttgtc tggcaaagag
     1201 gccaaggacg cagatggcaa caaacccaag gaacctggga taacggaggc ttcatcggag
     1261 atgatatcac caaacatgtt gctggtgatt ataataccat ttaggtgggt tgggttctta
     1321 actaggatca tggcggcaga atcaatcaat tgatgttgaa ccttcaatgt aggaaattcg
     1381 ttcttgatgg tttcctccac agtttttctc cataatcttg aagaggccaa aacattagct
     1441 ttatccaagg accaaatagg caatggtggc tcatgttgta gggccatgaa agcggccatt
     1501 cttgtgattc tttgcacttc tggaacggtg tattgttcac tatcccaagc gacaccatca
     1561 ccatcatctt cctttctctt accaaagtaa atacctccca ctaattctct gacaacaacg
     1621 aagtcagtac ctttagcaaa ttgtggcttg attggagata agtctaaaag agagtcggat
     1681 gcaaagttac atggtcttaa gttggcgtac aattgaagtt ctttacggat ttttagtaaa
     1741 ccttgttcag gtctaacact acctgtaccc catttaggac cacccacagc acctaacaaa
     1801 acggcatcag ccttcttgga ggcttccagc gcctcatctg gaagtgggac acctgtagct
     1861 tcgatagcag caccaccaat taaatgattt tcgaaatcga acttgacatt ggaacgaaca
     1921 tcagaaatag ctttaagaac cttaatggct tcggctgtga tttcttgacc aacgtggtca
     1981 cctggcaaaa cgacgatctt cttaggggca gacattagaa tggtatatcc ttgaaatata
     2041 tatatatatt gctgaaatgt aaaaggtaag aaaagttaga aagtaagacg attgctaacc
     2101 acctattgga aaaacaatag gtccttaaat aatattgtca acttcaagta ttgtgatgca
     2161 agcatttagt catgaacgct tctctattct atatgaaaag ccggttccgg cgctctcacc
     2221 tttccttttt ctcccaattt ttcagttgaa aaaggtatat gcgtcaggcg acctctgaaa
     2281 ttaacaaaaa atttccagtc atcgaatttg attctgtgcg atagcgcccc tgtgtgttct
     2341 cgttatgttg aggaaaaaaa taatggttaa taacttcgta taatgtatgc tatacgaagt
     2401 tatcccgggt accgagctcg aattcatcac gtgctataaa aataattata atttaaattt
     2461 tttaatataa atatataaat taaaaataga aagtaaaaaa agaaattaaa gaaaaaatag
     2521 tttttgtttt ccgaagatgt aaaagactct agggggatcg ccaacaaata ctacctttta
     2581 tcttgctctt cctgctctca ggtattaatg ccgaattgtt tcatcttgtc tgtgtagaag
     2641 accacacacg aaaatcctgt gattttacat tttacttatc gttaatcgaa tgtatatcta
     2701 tttaatctgc ttttcttgtc taataaatat atatgtaaag tacgcttttt gttgaaattt
     2761 tttaaacctt tgtttatttt tttttcttca ttccgtaact cttctacctt ctttatttac
     2821 tttctaaaat ccaaatacaa aacataaaaa taaataaaca cagagtaaat tcccaaatta
     2881 ttccatcatt aaaagatacg aggcgcgtgt aagttacgaa ttcgtaatca tggtcatagc
     2941 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     3001 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     3061 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     3121 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     3181 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     3241 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     3301 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3361 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3421 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3481 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     3541 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3601 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3661 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3721 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     3781 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3841 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3901 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3961 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4021 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     4081 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4141 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4201 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4261 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     4321 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4381 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     4441 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     4501 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     4561 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     4621 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4681 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     4741 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     4801 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4861 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4921 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     4981 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     5041 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     5101 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     5161 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     5221 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     5281 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcac
//
