LOCUS       sequence-12705-e        5121 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 5121)
  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 5121)
  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: pXP318
FEATURES             Location/Qualifiers
     source          1..5121
                     /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)."
     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"
     promoter        4233..4646
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      4684..4931
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4700..4718)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     5094..5112
                     /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 aattcatcac gtgctataaa aataattata atttaaattt
     1681 tttaatataa atatataaat taaaaataga aagtaaaaaa agaaattaaa gaaaaaatag
     1741 tttttgtttt ccgaagatgt aaaagactct agggggatcg ccaacaaata ctacctttta
     1801 tcttgctctt cctgctctca ggtattaatg ccgaattgtt tcatcttgtc tgtgtagaag
     1861 accacacacg aaaatcctgt gattttacat tttacttatc gttaatcgaa tgtatatcta
     1921 tttaatctgc ttttcttgtc taataaatat atatgtaaag tacgcttttt gttgaaattt
     1981 tttaaacctt tgtttatttt tttttcttca ttccgtaact cttctacctt ctttatttac
     2041 tttctaaaat ccaaatacaa aacataaaaa taaataaaca cagagtaaat tcccaaatta
     2101 ttccatcatt aaaagatacg aggcgcgtgt aagttacgaa 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 tattaccgcg
     4201 aatccttaca tcacacccaa tcccccacaa gtgatccccc acacaccata gcttcaaaat
     4261 gtttctactc cttttttact cttccagatt ttctcggact ccgcgcatcg ccgtaccact
     4321 tcaaaacacc caagcacagc atactaaatt tcccctcttt cttcctctag ggtgtcgtta
     4381 attacccgta ctaaaggttt ggaaaagaaa aaagagaccg cctcgtttct ttttcttcgt
     4441 cgaaaaaggc aataaaaatt tttatcacgt ttctttttct tgaaaatttt tttttttgat
     4501 ttttttctct ttcgatgacc tcccattgat atttaagtta ataaacggtc ttcaatttct
     4561 caagtttcag tttcattttt cttgttctat tacaactttt tttacttctt gctcattaga
     4621 aagaaagcat agcaatctaa tctaagtttt aattacaaaa ctagtgatat ctgcgcactc
     4681 gagtcatgta attagttatg tcacgcttac attcacgccc tccccccaca tccgctctaa
     4741 ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt tttatagtta
     4801 tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg tacagacgcg
     4861 tgtacgcatg taacattata ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg
     4921 ctttaatttg cggccaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     4981 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5041 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     5101 gtatcacgag gccctttcgt c
//
