LOCUS       sequence-12699-e        6226 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 6226)
  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 6226)
  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: pXP218
FEATURES             Location/Qualifiers
     source          1..6226
                     /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)."
     CDS             complement(552..1355)
                     /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"
     promoter        complement(1356..1572)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     protein_bind    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      1650..2992
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    1982..2029
                     /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"
     terminator      complement(5054..5301)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     5267..5285
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     6199..6217
                     /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 tatgggtaat
      481 aactgatata attaaattga agctctaatt tgtgagttta gtatacatgc atttacttat
      541 aatacagttt tttagttttg ctggccgcat cttctcaaat atgcttccca gcctgctttt
      601 ctgtaacgtt caccctctac cttagcatcc cttccctttg caaatagtcc tcttccaaca
      661 ataataatgt cagatcctgt agagaccaca tcatccacgg ttctatactg ttgacccaat
      721 gcgtctccct tgtcatctaa acccacaccg ggtgtcataa tcaaccaatc gtaaccttca
      781 tctcttccac ccatgtctct ttgagcaata aagccgataa caaaatcttt gtcgctcttc
      841 gcaatgtcaa cagtaccctt agtatattct ccagtagcta gggagccctt gcatgacaat
      901 tctgctaaca tcaaaaggcc tctaggttcc tttgttactt cttccgccgc ctgcttcaaa
      961 ccgctaacaa tacctgggcc caccacaccg tgtgcattcg taatgtctgc ccattctgct
     1021 attctgtata cacccgcaga gtactgcaat ttgactgtat taccaatgtc agcaaatttt
     1081 ctgtcttcga agagtaaaaa attgtacttg gcggataatg cctttagcgg cttaactgtg
     1141 ccctccatgg aaaaatcagt caagatatcc acatgtgttt ttagtaaaca aattttggga
     1201 cctaatgctt caactaactc cagtaattcc ttggtggtac gaacatccaa tgaagcacac
     1261 aagtttgttt gcttttcgtg catgatatta aatagcttgg cagcaacagg actaggatga
     1321 gtagcagcac gttccttata tgtagctttc gacatgattt atcttcgttt cctgcaggtt
     1381 tttgttctgt gcagttgggt taagaatact gggcaatttc atgtttcttc aacaccacat
     1441 atgcgtatat ataccaatct aagtctgtgc tccttccttc gttcttcctt ctgctcggag
     1501 attaccgaat caaaaaaatt tcaaagaaac cggaatcaaa aaaaagaaca aaaaaaaaaa
     1561 agatgaattg aaaagcttta tggaccctga taacttcgta taatgtatgc tatacgaagt
     1621 tatcccgggt accgagctcg aattcgtatg atccaatatc aaaggaaatg atagcattga
     1681 aggatgagac taatccaatt gaggagtggc agcatataga acagctaaag ggtagtgctg
     1741 aaggaagcat acgatacccc gcatggaatg ggataatatc acaggaggta ctagactacc
     1801 tttcatccta cataaataga cgcatataag tacgcattta agcataaaca cgcactatgc
     1861 cgttcttctc atgtatatat atatacaggc aacacgcaga tataggtgcg acgtgaacag
     1921 tgagctgtat gtgcgcagct cgcgttgcat tttcggaagc gctcgttttc ggaaacgctt
     1981 tgaagttcct attccgaagt tcctattctc tagaaagtat aggaacttca gagcgctttt
     2041 gaaaaccaaa agcgctctga agacgcactt tcaaaaaacc aaaaacgcac cggactgtaa
     2101 cgagctacta aaatattgcg aataccgctt ccacaaacat tgctcaaaag tatctctttg
     2161 ctatatatct ctgtgctata tccctatata acctacccat ccacctttcg ctccttgaac
     2221 ttgcatctaa actcgacctc tacatttttt atgtttatct ctagtattac tctttagaca
     2281 aaaaaattgt agtaagaact attcatagag tgaatcgaaa acaatacgaa aatgtaaaca
     2341 tttcctatac gtagtatata gagacaaaat agaagaaacc gttcataatt ttctgaccaa
     2401 tgaagaatca tcaacgctat cactttctgt tcacaaagta tgcgcaatcc acatcggtat
     2461 agaatataat cggggatgcc tttatcttga aaaaatgcac ccgcagcttc gctagtaatc
     2521 agtaaacgcg ggaagtggag tcaggctttt tttatggaag agaaaataga caccaaagta
     2581 gccttcttct aaccttaacg gacctacagt gcaaaaagtt atcaagagac tgcattatag
     2641 agcgcacaaa ggagaaaaaa agtaatctaa gatgctttgt tagaaaaata gcgctctcgg
     2701 gatgcatttt tgtagaacaa aaaagaagta tagattcttt gttggtaaaa tagcgctctc
     2761 gcgttgcatt tctgttctgt aaaaatgcag ctcagattct ttgtttgaaa aattagcgct
     2821 ctcgcgttgc atttttgttt tacaaaaatg aagcacagat tcttcgttgg taaaatagcg
     2881 ctttcgcgtt gcatttctgt tctgtaaaaa tgcagctcag attctttgtt tgaaaaatta
     2941 gcgctctcgc gttgcatttt tgttctacaa aatgaagcac agatgcttcg ttgaattcgt
     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 ttcaatattg gccgcaaatt aaagccttcg agcgtcccaa aaccttctca agcaaggttt
     5101 tcagtataat gttacatgcg tacacgcgtc tgtacagaaa aaaaagaaaa atttgaaata
     5161 taaataacgt tcttaatact aacataacta taaaaaaata aatagggacc tagacttcag
     5221 gttgtctaac tccttccttt tcggttagag cggatgtggg gggagggcgt gaatgtaagc
     5281 gtgacataac taattacatg actcgagtgc gcagatatca ctagtatatt tgttgtaaaa
     5341 agtagataat tacttccttg atgatctgta aaaaagagaa aaagaaagca tctaagaact
     5401 tgaaaaacta cgaattagaa aagaccaaat atgtatttct tgcattgacc aatttatgca
     5461 agtttatata tatgtaaatg taagtttcac gaggttctac taaactaaac cacccccttg
     5521 gttagaagaa aagagtgtgt gagaacaggc tgttgttgtc acacgattcg gacaattctg
     5581 tttgaaagag agagagtaac agtacgatcg aacgaacttt gctctggaga tcacagtggg
     5641 catcatagca tgtggtacta aaccctttcc cgccattcca gaaccttcga ttgcttgtta
     5701 caaaacctgt gagccgtcgc taggaccttg ttgtgtgacg aaattggaag ctgcaatcaa
     5761 taggaagaca ggaagtcgag cgtgtctggg ttttttcagt tttgttcttt ttgcaaacaa
     5821 atcacgagcg acggtaattt ctttctcgat aagaggccac gtgctttatg agggtaacat
     5881 caattcaaga aggagggaaa cacttccttt ttctggccct gataatagta tgagggtgaa
     5941 gccaaaataa aggattcgcg cccaaatcgg catctttaaa tgcaggtatg cgatagttcc
     6001 tcactctttc cttactcacg agtaattctt gcaaatgcct aatattattg aagcatttat
     6061 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6121 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     6181 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtc
//
