LOCUS       sequence-65296-e        5866 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 5866)
  AUTHORS   Shen MW, Fang F, Sandmeyer S, Da Silva NA
  TITLE     Development and characterization of a vector set with regulated 
            promoters for systematic metabolic engineering in Saccharomyces 
            cerevisiae.
  JOURNAL   Yeast. 2012 Dec;29(12):495-503. doi: 10.1002/yea.2930. Epub 2012 Nov
            19.
  PUBMED    23166051
REFERENCE   2  (bases 1 to 5866)
  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.2930"; 
            journalName: "Yeast"; date: "2012-12"; volume: "29"; issue: "12"; 
            pages: "495-503"
COMMENT     Sequence Label: pXP842
FEATURES             Location/Qualifiers
     source          1..5866
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      602..849
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(618..636)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     protein_bind    complement(895..928)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        946..1162
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             1163..1966
                     /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(2045..2078)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      2101..3443
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(3064..3111)
                     /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(3463..3479)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3463..3479)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3476..3498)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3487..3503
                     /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(3511..3517,3518..3535,3536..3541))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3556..3577
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3694..3711)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3865..4453)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3945..3964)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4624..5415,5416..5484))
                     /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     5247..5266
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5485..5589)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5657..5675
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5713..5735)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5835..5854
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 catatgattt agcggccgca aaacgtaggg gcaaacaaac ggaaaaatcg tttctcaaat
       61 tttctgatgc caagaactct aaccagtctt atctaaaaat tgccttatga tccgtctctc
      121 cggttacagc ctgtgtaact gattaatcct gcctttctaa tcaccattct aatgttttaa
      181 ttaagggatt ttgtcttcat taacggcttt cgctcataaa aatgttatga cgttttgccc
      241 gcaggcggga aaccatccac ttcacgagac tgatctcctc tgccggaaca ccgggcatct
      301 ccaacttata agttggagaa ataagagaat ttcagattga gagaatgaaa aaaaaaaaaa
      361 aaaaaaaggc agaggagagc atagaaatgg ggttcacttt ttggtaaagc tatagcatgc
      421 ctatcacata taaatagagt gccagtagcg acttttttca cactcgaaat actcttacta
      481 ctgctctctt gttgttttta tcacttcttg tttcttcttg gtaaatagaa tatcaagcta
      541 caaaaagcat acaatcaact atcaactatt aactatatcg taatacacaa ctagtctcga
      601 gtcatgtaat tagttatgtc acgcttacat tcacgccctc cccccacatc cgctctaacc
      661 gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt tatagttatg
      721 ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta cagacgcgtg
      781 tacgcatgta acattatact gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct
      841 ttaatttgcg gccaagcttg catgcctgca ggtcgactct agaggatccc cgggataact
      901 tcgtatagca tacattatac gaagttatca gggtccataa agcttttcaa ttcatctttt
      961 ttttttttgt tctttttttt gattccggtt tctttgaaat ttttttgatt cggtaatctc
     1021 cgagcagaag gaagaacgaa ggaaggagca cagacttaga ttggtatata tacgcatatg
     1081 tggtgttgaa gaaacatgaa attgcccagt attcttaacc caactgcaca gaacaaaaac
     1141 ctgcaggaaa cgaagataaa tcatgtcgaa agctacatat aaggaacgtg ctgctactca
     1201 tcctagtcct gttgctgcca agctatttaa tatcatgcac gaaaagcaaa caaacttgtg
     1261 tgcttcattg gatgttcgta ccaccaagga attactggag ttagttgaag cattaggtcc
     1321 caaaatttgt ttactaaaaa cacatgtgga tatcttgact gatttttcca tggagggcac
     1381 agttaagccg ctaaaggcat tatccgccaa gtacaatttt ttactcttcg aagacagaaa
     1441 atttgctgac attggtaata cagtcaaatt gcagtactct gcgggtgtat acagaatagc
     1501 agaatgggca gacattacga atgcacacgg tgtggtgggc ccaggtattg ttagcggttt
     1561 gaagcaggcg gcggaagaag taacaaagga acctagaggc cttttgatgt tagcagaatt
     1621 gtcatgcaag ggctccctag ctactggaga atatactaag ggtactgttg acattgcgaa
     1681 gagcgacaaa gattttgtta tcggctttat tgctcaaaga gacatgggtg gaagagatga
     1741 aggttacgat tggttgatta tgacacccgg tgtgggttta gatgacaagg gagacgcatt
     1801 gggtcaacag tatagaaccg tggatgatgt ggtctctaca ggatctgaca ttattattgt
     1861 tggaagagga ctatttgcaa agggaaggga tgctaaggta gagggtgaac gttacagaaa
     1921 agcaggctgg gaagcatatt tgagaagatg cggccagcaa aactaaaaaa ctgtattata
     1981 agtaaatgca tgtatactaa actcacaaat tagagcttca atttaattat atcagttatt
     2041 acccataact tcgtatagca tacattatac gaagttatcc cgggtaccga gctcgaattc
     2101 aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc gctaattttt
     2161 caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa agcgctattt
     2221 taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa
     2281 tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgagagcgc
     2341 tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca tcccgagagc
     2401 gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg ctctataatg
     2461 cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag gctactttgg
     2521 tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta ctgattacta
     2581 gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt ctataccgat
     2641 gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt cattggtcag
     2701 aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga aatgtttaca
     2761 ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt tttgtctaaa
     2821 gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc aagttcaagg
     2881 agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat agcaaagaga
     2941 tacttttgag caatgtttgt ggaagcggta ttcgcaatat tttagtagct cgttacagtc
     3001 cggtgcgttt ttggtttttt gaaagtgcgt cttcagagcg cttttggttt tcaaaagcgc
     3061 tctgaagttc ctatactttc tagagaatag gaacttcgga ataggaactt caaagcgttt
     3121 ccgaaaacga gcgcttccga aaatgcaacg cgagctgcgc acatacagct cactgttcac
     3181 gtcgcaccta tatctgcgtg ttgcctgtat atatatatac atgagaagaa cggcatagtg
     3241 cgtgtttatg cttaaatgcg tacttatatg cgtctattta tgtaggatga aaggtagtct
     3301 agtacctcct gtgatattat cccattccat gcggggtatc gtatgcttcc ttcagcacta
     3361 ccctttagct gttctatatg ctgccactcc tcaattggat tagtctcatc cttcaatgct
     3421 atcatttcct ttgatattgg atcatacgaa ttcgtaatca tggtcatagc tgtttcctgt
     3481 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     3541 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     3601 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     3661 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     3721 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3781 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3841 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3901 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3961 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4021 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4081 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4141 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4201 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4261 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     4321 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4381 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4441 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4501 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4561 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4621 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4681 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4741 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4801 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4861 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4921 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4981 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5041 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5101 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5161 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5221 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5281 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5341 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5401 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5461 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     5521 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     5581 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     5641 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     5701 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     5761 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     5821 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcac
//
