LOCUS       sequence-65295-e        6191 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 6191)
  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 6191)
  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: pXP841
FEATURES             Location/Qualifiers
     source          1..6191
                     /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)."
     rep_origin      2426..3768
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(3389..3436)
                     /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(3788..3804)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3788..3804)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3801..3823)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3812..3828
                     /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(3836..3842,3843..3860,3861..3866))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3881..3902
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4019..4036)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4190..4778)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4270..4289)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4949..5740,5741..5809))
                     /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     5572..5591
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5810..5914)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5982..6000
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6038..6060)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6160..6179
                     /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 aattcaacga agcatctgtg cttcattttg tagaacaaaa
     2461 atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca
     2521 gaaatgcaac gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa
     2581 caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca
     2641 gaacagaaat gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg
     2701 ttctacaaaa atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt
     2761 tttctccttt gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt
     2821 taaggttaga agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc
     2881 cacttcccgc gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc
     2941 atccccgatt atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata
     3001 gcgttgatga ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat
     3061 atactacgta taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt
     3121 tcttactaca atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag
     3181 gtcgagttta gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat
     3241 agcacagaga tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc
     3301 aatattttag tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcttca
     3361 gagcgctttt ggttttcaaa agcgctctga agttcctata ctttctagag aataggaact
     3421 tcggaatagg aacttcaaag cgtttccgaa aacgagcgct tccgaaaatg caacgcgagc
     3481 tgcgcacata cagctcactg ttcacgtcgc acctatatct gcgtgttgcc tgtatatata
     3541 tatacatgag aagaacggca tagtgcgtgt ttatgcttaa atgcgtactt atatgcgtct
     3601 atttatgtag gatgaaaggt agtctagtac ctcctgtgat attatcccat tccatgcggg
     3661 gtatcgtatg cttccttcag cactaccctt tagctgttct atatgctgcc actcctcaat
     3721 tggattagtc tcatccttca atgctatcat ttcctttgat attggatcat acgaattcgt
     3781 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     3841 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     3901 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     3961 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4021 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4081 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4141 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4201 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4261 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4321 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4381 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4441 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4501 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4561 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4621 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4681 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4741 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4801 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4861 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4921 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     4981 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5041 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5101 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5161 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5221 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5281 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5341 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5401 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5461 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5521 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     5581 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     5641 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     5701 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     5761 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     5821 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     5881 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     5941 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     6001 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     6061 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     6121 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     6181 tgagagtgca c
//
