LOCUS       sequence-12711-e        5976 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 5976)
  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 5976)
  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: pXP418
FEATURES             Location/Qualifiers
     source          1..5976
                     /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)."
     promoter        491..707
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             708..1511
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLATGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     protein_bind    complement(1590..1623)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      1646..2988
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(2609..2656)
                     /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(3008..3024)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3008..3024)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3021..3043)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3032..3048
                     /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(3056..3062,3063..3080,3081..3086))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3101..3122
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3239..3256)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3410..3998)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3490..3509)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4169..4960,4961..5029))
                     /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     4792..4811
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        5088..5501
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      5539..5786
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5555..5573)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     5949..5967
                     /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 tatcagggtc
      481 cataaagctt ttcaattcat cttttttttt tttgttcttt tttttgattc cggtttcttt
      541 gaaatttttt tgattcggta atctccgagc agaaggaaga acgaaggaag gagcacagac
      601 ttagattggt atatatacgc atatgtggtg ttgaagaaac atgaaattgc ccagtattct
      661 taacccaact gcacagaaca aaaacctgca ggaaacgaag ataaatcatg tcgaaagcta
      721 catataagga acgtgctgct actcatccta gtcctgttgc tgccaagcta tttaatatca
      781 tgcacgaaaa gcaaacaaac ttgtgtgctt cattggatgt tcgtaccacc aaggaattac
      841 tggagttagt tgaagcatta ggtcccaaaa tttgtttact aaaaacacat gtggatatct
      901 tgactgattt ttccatggag ggcacagtta agccgctaaa ggcattatcc gccaagtaca
      961 attttttact cttcgaagac agaaaatttg ctgacattgg taatacagtc aaattgcagt
     1021 actctgcggg tgtatacaga atagcagaat gggcagacat tacgaatgca cacggtgtgg
     1081 tgggcccagg tattgttagc ggtttgaagc aggcggcgga agaagtaaca aaggaaccta
     1141 gaggcctttt gatgttagca gaattgtcat gcaagggctc cctagctact ggagaatata
     1201 ctaagggtac tgttgacatt gcgaagagcg acaaagattt tgttatcggc tttattgctc
     1261 aaagagacat gggtggaaga gatgaaggtt acgattggtt gattatgaca cccggtgtgg
     1321 gtttagatga caagggagac gcattgggtc aacagtatag aaccgtggat gatgtggtct
     1381 ctacaggatc tgacattatt attgttggaa gaggactatt tgcaaaggga agggatgcta
     1441 aggtagaggg tgaacgttac agaaaagcag gctgggaagc atatttgaga agatgcggcc
     1501 agcaaaacta aaaaactgta ttataagtaa atgcatgtat actaaactca caaattagag
     1561 cttcaattta attatatcag ttattaccca taacttcgta tagcatacat tatacgaagt
     1621 tatcccgggt accgagctcg aattcaacga agcatctgtg cttcattttg tagaacaaaa
     1681 atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca
     1741 gaaatgcaac gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa
     1801 caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca
     1861 gaacagaaat gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg
     1921 ttctacaaaa atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt
     1981 tttctccttt gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt
     2041 taaggttaga agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc
     2101 cacttcccgc gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc
     2161 atccccgatt atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata
     2221 gcgttgatga ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat
     2281 atactacgta taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt
     2341 tcttactaca atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag
     2401 gtcgagttta gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat
     2461 agcacagaga tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc
     2521 aatattttag tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcttca
     2581 gagcgctttt ggttttcaaa agcgctctga agttcctata ctttctagag aataggaact
     2641 tcggaatagg aacttcaaag cgtttccgaa aacgagcgct tccgaaaatg caacgcgagc
     2701 tgcgcacata cagctcactg ttcacgtcgc acctatatct gcgtgttgcc tgtatatata
     2761 tatacatgag aagaacggca tagtgcgtgt ttatgcttaa atgcgtactt atatgcgtct
     2821 atttatgtag gatgaaaggt agtctagtac ctcctgtgat attatcccat tccatgcggg
     2881 gtatcgtatg cttccttcag cactaccctt tagctgttct atatgctgcc actcctcaat
     2941 tggattagtc tcatccttca atgctatcat ttcctttgat attggatcat acgaattcgt
     3001 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     3061 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     3121 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     3181 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     3241 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     3301 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     3361 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3421 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3481 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3541 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     3601 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3661 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3721 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     3781 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     3841 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     3901 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     3961 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4021 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4081 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4141 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     4201 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     4261 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     4321 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     4381 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     4441 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     4501 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     4561 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     4621 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     4681 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     4741 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     4801 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     4861 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     4921 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     4981 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     5041 ttcaatatta ccgcgaatcc ttacatcaca cccaatcccc cacaagtgat cccccacaca
     5101 ccatagcttc aaaatgtttc tactcctttt ttactcttcc agattttctc ggactccgcg
     5161 catcgccgta ccacttcaaa acacccaagc acagcatact aaatttcccc tctttcttcc
     5221 tctagggtgt cgttaattac ccgtactaaa ggtttggaaa agaaaaaaga gaccgcctcg
     5281 tttctttttc ttcgtcgaaa aaggcaataa aaatttttat cacgtttctt tttcttgaaa
     5341 attttttttt ttgatttttt tctctttcga tgacctccca ttgatattta agttaataaa
     5401 cggtcttcaa tttctcaagt ttcagtttca tttttcttgt tctattacaa ctttttttac
     5461 ttcttgctca ttagaaagaa agcatagcaa tctaatctaa gttttaatta caaaactagt
     5521 gatatctgcg cactcgagtc atgtaattag ttatgtcacg cttacattca cgccctcccc
     5581 ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag gtccctattt
     5641 atttttttat agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt
     5701 ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt gagaaggttt
     5761 tgggacgctc gaaggcttta atttgcggcc aatattattg aagcatttat cagggttatt
     5821 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     5881 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     5941 cctataaaaa taggcgtatc acgaggccct ttcgtc
//
