LOCUS       sequence-12693-e        5371 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 5371)
  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 5371)
  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: pXP118
FEATURES             Location/Qualifiers
     source          1..5371
                     /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)."
     primer_bind     complement(2153..2169)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2153..2169)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2166..2188)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2177..2193
                     /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(2201..2207,2208..2225,2226..2231))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2246..2267
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2384..2401)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2555..3143)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2635..2654)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3314..4105,4106..4174))
                     /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     3937..3956
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      complement(4199..4446)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     4412..4430
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     5344..5362
                     /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 aattcgtaac ttacacgcgc ctcgtatctt ttaatgatgg
     1681 aataatttgg gaatttactc tgtgtttatt tatttttatg ttttgtattt ggattttaga
     1741 aagtaaataa agaaggtaga agagttacgg aatgaagaaa aaaaaataaa caaaggttta
     1801 aaaaatttca acaaaaagcg tactttacat atatatttat tagacaagaa aagcagatta
     1861 aatagatata cattcgatta acgataagta aaatgtaaaa tcacaggatt ttcgtgtgtg
     1921 gtcttctaca cagacaagat gaaacaattc ggcattaata cctgagagca ggaagagcaa
     1981 gataaaaggt agtatttgtt ggcgatcccc ctagagtctt ttacatcttc ggaaaacaaa
     2041 aactattttt tctttaattt ctttttttac tttctatttt taatttatat atttatatta
     2101 aaaaatttaa attataatta tttttatagc acgtgatgaa ttcgtaatca tggtcatagc
     2161 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     2221 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     2281 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     2341 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     2401 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     2461 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     2521 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2581 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2641 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2701 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     2761 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     2821 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     2881 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     2941 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     3001 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3061 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3121 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3181 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     3241 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3301 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3361 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     3421 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     3481 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     3541 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     3601 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     3661 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     3721 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     3781 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     3841 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     3901 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     3961 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     4021 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4081 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4141 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattggccgc
     4201 aaattaaagc cttcgagcgt cccaaaacct tctcaagcaa ggttttcagt ataatgttac
     4261 atgcgtacac gcgtctgtac agaaaaaaaa gaaaaatttg aaatataaat aacgttctta
     4321 atactaacat aactataaaa aaataaatag ggacctagac ttcaggttgt ctaactcctt
     4381 ccttttcggt tagagcggat gtggggggag ggcgtgaatg taagcgtgac ataactaatt
     4441 acatgactcg agtgcgcaga tatcactagt atatttgttg taaaaagtag ataattactt
     4501 ccttgatgat ctgtaaaaaa gagaaaaaga aagcatctaa gaacttgaaa aactacgaat
     4561 tagaaaagac caaatatgta tttcttgcat tgaccaattt atgcaagttt atatatatgt
     4621 aaatgtaagt ttcacgaggt tctactaaac taaaccaccc ccttggttag aagaaaagag
     4681 tgtgtgagaa caggctgttg ttgtcacacg attcggacaa ttctgtttga aagagagaga
     4741 gtaacagtac gatcgaacga actttgctct ggagatcaca gtgggcatca tagcatgtgg
     4801 tactaaaccc tttcccgcca ttccagaacc ttcgattgct tgttacaaaa cctgtgagcc
     4861 gtcgctagga ccttgttgtg tgacgaaatt ggaagctgca atcaatagga agacaggaag
     4921 tcgagcgtgt ctgggttttt tcagttttgt tctttttgca aacaaatcac gagcgacggt
     4981 aatttctttc tcgataagag gccacgtgct ttatgagggt aacatcaatt caagaaggag
     5041 ggaaacactt cctttttctg gccctgataa tagtatgagg gtgaagccaa aataaaggat
     5101 tcgcgcccaa atcggcatct ttaaatgcag gtatgcgata gttcctcact ctttccttac
     5161 tcacgagtaa ttcttgcaaa tgcctaatat tattgaagca tttatcaggg ttattgtctc
     5221 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     5281 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     5341 aaaaataggc gtatcacgag gccctttcgt c
//
