LOCUS       sequence-12696-e        7837 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 7837)
  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 7837)
  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: pXP212
FEATURES             Location/Qualifiers
     source          1..7837
                     /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    440..473
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3201..3234
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      3261..4603
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    3593..3640
                     /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(4619..4635)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4619..4635)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4632..4654)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4643..4659
                     /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(4667..4673,4674..4691,4692..4697))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4712..4733
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4850..4867)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5021..5609)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5101..5120)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5780..6571,6572..6640))
                     /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     6403..6422
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      complement(6665..6912)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     6878..6896
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     7810..7828
                     /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 taatgtatgc tatacgaagt tatggtgaat
      481 catcgataaa aataaatata ctgagattat agtaagctca ttgatccctt aaactttctt
      541 ttcggtgtat gacttatgag ggtgagaatg cgaaatggcg tggaaatgtg atcaaaggta
      601 ataaaacgtc atatctatgc tacaacattc caaaatttgt cccaaaaagt ctttggttca
      661 tgatcttccc atacaattgc ctcaatgtct cttctatcgg aatcgatgtc gacatctcca
      721 atcttccaaa taaatctgca tctgaatatg cattgaaata agatccaaac agctaagaac
      781 aggaaaatag agatataggc ggcagcaaag ctaacaccat tgaattttgg tgcaaaagcc
      841 gtgaaacctt gaataatgat aatgatcgtc ataaatgtgg ccgcataata agccaagccg
      901 ggcattaatt tagctttaaa tggtaactcg tcacgagaga tgccacggta tttcaaagct
      961 tgcataaatc tgatgtgcga gattgagata aataaccatg caaaaaagcc tgcaacacca
     1021 gtgatattta atagccattc gaaaactttg tcaccaccag tagatgtctc catgtaagcc
     1081 aaagcgccaa atgcagcagt aacgaaaact gcaatgtatg gaacaccacc tttggtggtc
     1141 cttgacagga atttaggagc caacttgttc tttgatagac caaataaaat acgggaacca
     1201 acgtaaatat ttgaatttgc ggcagaaata atggttgtta agataacagc gttgaagata
     1261 tgtggcaaaa cctttgtacc agagttctca atagcaataa taaagggaga agtagaaacg
     1321 taggaagtag attgtgttag tttagggtca ttgtatggaa ctaaaagtcc aatgaataat
     1381 agagagccaa tgtagaaggt taagatacgg aaaacaactt ttttgatggc tcttggaacg
     1441 gattttctgg ggtttgcagc ttcaccagca gtgataccaa cgagttcagt accttgaaat
     1501 gtgaaggcag cgttaatcaa agaggaaacc caacctaaga acctcccttc gtttttatcc
     1561 ttagatatta tacctggacc ccaggcacct gggtttctcc aataacggaa tccaactggg
     1621 ccggtaaccc cagcaccaca aaccatacaa aaacagtata ttagaaaccc gataatggct
     1681 aaaactttga tggaagcgac ccagaactcg aattcaccgt aatatttgac agggaacaag
     1741 ttcattattg tgataattac ccaaaaaata ctaatccatg ccgccagtgg aactttgtac
     1801 gtccaaaatt gaatgacttg gccaactaca ctaagttcca gggcaaaagt gattgcccaa
     1861 gaaaaccaat acatgtaacc attggccgca ccaaatgctg gagaaaggaa tctttgtgag
     1921 aaaactgtga aagaggatgt aacagggatg aatgtagcca tttcacccaa ggactgcgtg
     1981 acagaatatg ccaaagaacc cataaataaa tatgatataa gagcgcccac tgggccggcg
     2041 ttggtcagag gtgtggataa accaatgaaa agacctgtac caatagtacc accaagggca
     2101 atcataccaa tatgtctttg cttaagctct ctcttcactt cagcgttctg tacttctcct
     2161 tcatcttcat cacctatgcc atcctccata gagaacgtat cctcgccatt tactctcgtc
     2221 gggaaagagc gcaatggata caattcttta cttttctcat ctttcaatgg tattgaccca
     2281 cgtctgtggt gtgtttgtga agcttcaacg tcgtgaaaga gggttgtgac cggctcattg
     2341 tacatatgct tctcctctat gtcggcgtct tcttttgaat ttgtcattgc tatgcctttt
     2401 tttttttttg tttttacagg agttaagaag tctgaagaac tctgaaataa actttcgatt
     2461 gacgacagat taaaaactgg atccaactat aagtataata gtaacttata tatttctgtt
     2521 ccagatatat acgtaaaaaa aagaaaagga taattgatgt ttttgcaaaa agggattccc
     2581 tgattcggtt tactctgttc gcaaccactc ttctttctgc taatttgtgc tttatctcgt
     2641 atcattctct tatctactta tcttaacctt atcgtcttcc tcgaaatttt cattgataga
     2701 gacaactgca aaaaaagaaa ttcaaaaaag ccgtatgact cacccggaaa ccacacagaa
     2761 accctagatt tctatagggc aaatttcagg gttatactaa aagatatatt attgaccttg
     2821 gttattcttc aagttcgccc tattgaaaat tcggacactt tagggtttgg aatctgttaa
     2881 agatataagt aaatatgatg ataaagaaat aggtttaatt tatacgcact tgccagtaag
     2941 tgattgatat tgcagaagta acatgaatac ggttctgtgg cctttgctgt ttgccgttat
     3001 tttatttctt ttttttgttt tttttctttt tttcagcact atcaaggcta ataagggaca
     3061 agagttctta ctcagtgtga acgtgttcta aataagttct tgttctaatt aattgtaact
     3121 atttttatgc atttcttgta agaggaactg gagtaatcgt atctcggaaa tacatagagg
     3181 tgcgtattca taatgggccc ataacttcgt ataatgtatg ctatacgaag ttatcccggg
     3241 taccgagctc gaattcgtat gatccaatat caaaggaaat gatagcattg aaggatgaga
     3301 ctaatccaat tgaggagtgg cagcatatag aacagctaaa gggtagtgct gaaggaagca
     3361 tacgataccc cgcatggaat gggataatat cacaggaggt actagactac ctttcatcct
     3421 acataaatag acgcatataa gtacgcattt aagcataaac acgcactatg ccgttcttct
     3481 catgtatata tatatacagg caacacgcag atataggtgc gacgtgaaca gtgagctgta
     3541 tgtgcgcagc tcgcgttgca ttttcggaag cgctcgtttt cggaaacgct ttgaagttcc
     3601 tattccgaag ttcctattct ctagaaagta taggaacttc agagcgcttt tgaaaaccaa
     3661 aagcgctctg aagacgcact ttcaaaaaac caaaaacgca ccggactgta acgagctact
     3721 aaaatattgc gaataccgct tccacaaaca ttgctcaaaa gtatctcttt gctatatatc
     3781 tctgtgctat atccctatat aacctaccca tccacctttc gctccttgaa cttgcatcta
     3841 aactcgacct ctacattttt tatgtttatc tctagtatta ctctttagac aaaaaaattg
     3901 tagtaagaac tattcataga gtgaatcgaa aacaatacga aaatgtaaac atttcctata
     3961 cgtagtatat agagacaaaa tagaagaaac cgttcataat tttctgacca atgaagaatc
     4021 atcaacgcta tcactttctg ttcacaaagt atgcgcaatc cacatcggta tagaatataa
     4081 tcggggatgc ctttatcttg aaaaaatgca cccgcagctt cgctagtaat cagtaaacgc
     4141 gggaagtgga gtcaggcttt ttttatggaa gagaaaatag acaccaaagt agccttcttc
     4201 taaccttaac ggacctacag tgcaaaaagt tatcaagaga ctgcattata gagcgcacaa
     4261 aggagaaaaa aagtaatcta agatgctttg ttagaaaaat agcgctctcg ggatgcattt
     4321 ttgtagaaca aaaaagaagt atagattctt tgttggtaaa atagcgctct cgcgttgcat
     4381 ttctgttctg taaaaatgca gctcagattc tttgtttgaa aaattagcgc tctcgcgttg
     4441 catttttgtt ttacaaaaat gaagcacaga ttcttcgttg gtaaaatagc gctttcgcgt
     4501 tgcatttctg ttctgtaaaa atgcagctca gattctttgt ttgaaaaatt agcgctctcg
     4561 cgttgcattt ttgttctaca aaatgaagca cagatgcttc gttgaattcg taatcatggt
     4621 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     4681 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     4741 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4801 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4861 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4921 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4981 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5041 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5101 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5161 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5221 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5281 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5341 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5401 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5461 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5521 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5581 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5641 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5701 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5761 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     5821 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     5881 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5941 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6001 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6061 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6121 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6181 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6241 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6301 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6361 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6421 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6481 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6541 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6601 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6661 ggccgcaaat taaagccttc gagcgtccca aaaccttctc aagcaaggtt ttcagtataa
     6721 tgttacatgc gtacacgcgt ctgtacagaa aaaaaagaaa aatttgaaat ataaataacg
     6781 ttcttaatac taacataact ataaaaaaat aaatagggac ctagacttca ggttgtctaa
     6841 ctccttcctt ttcggttaga gcggatgtgg ggggagggcg tgaatgtaag cgtgacataa
     6901 ctaattacat gactcgagtg cgcagatatc actagtatat ttgttgtaaa aagtagataa
     6961 ttacttcctt gatgatctgt aaaaaagaga aaaagaaagc atctaagaac ttgaaaaact
     7021 acgaattaga aaagaccaaa tatgtatttc ttgcattgac caatttatgc aagtttatat
     7081 atatgtaaat gtaagtttca cgaggttcta ctaaactaaa ccaccccctt ggttagaaga
     7141 aaagagtgtg tgagaacagg ctgttgttgt cacacgattc ggacaattct gtttgaaaga
     7201 gagagagtaa cagtacgatc gaacgaactt tgctctggag atcacagtgg gcatcatagc
     7261 atgtggtact aaaccctttc ccgccattcc agaaccttcg attgcttgtt acaaaacctg
     7321 tgagccgtcg ctaggacctt gttgtgtgac gaaattggaa gctgcaatca ataggaagac
     7381 aggaagtcga gcgtgtctgg gttttttcag ttttgttctt tttgcaaaca aatcacgagc
     7441 gacggtaatt tctttctcga taagaggcca cgtgctttat gagggtaaca tcaattcaag
     7501 aaggagggaa acacttcctt tttctggccc tgataatagt atgagggtga agccaaaata
     7561 aaggattcgc gcccaaatcg gcatctttaa atgcaggtat gcgatagttc ctcactcttt
     7621 ccttactcac gagtaattct tgcaaatgcc taatattatt gaagcattta tcagggttat
     7681 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7741 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     7801 acctataaaa ataggcgtat cacgaggccc tttcgtc
//
