LOCUS       sequence-12710-e        5865 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 5865)
  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 5865)
  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: pXP416
FEATURES             Location/Qualifiers
     source          1..5865
                     /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        474..755
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             756..1430
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     protein_bind    complement(1479..1512)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      1535..2877
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(2498..2545)
                     /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(2897..2913)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2897..2913)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2910..2932)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2921..2937
                     /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(2945..2951,2952..2969,2970..2975))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2990..3011
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3128..3145)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3299..3887)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3379..3398)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4058..4849,4850..4918))
                     /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     4681..4700
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        4977..5390
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      5428..5675
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5444..5462)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     5838..5856
                     /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 tataacgaca
      481 ttactatata tataatatag gaagcattta atagaacagc atcgtaatat atgtgtactt
      541 tgcagttatg acgccagatg gcagtagtgg aagatattct ttattgaaaa atagcttgtc
      601 accttacgta caatcttgat ccggagcttt tctttttttg ccgattaaga attaattcgg
      661 tcgaaaaaag aaaaggagag ggccaagagg gagggcattg gtgactattg agcacgtgag
      721 tatacgtgat taagcacaca aaggcagctt ggagtatgtc tgttattaat ttcacaggta
      781 gttctggtcc attggtgaaa gtttgcggct tgcagagcac agaggccgca gaatgtgctc
      841 tagattccga tgctgacttg ctgggtatta tatgtgtgcc caatagaaag agaacaattg
      901 acccggttat tgcaaggaaa atttcaagtc ttgtaaaagc atataaaaat agttcaggca
      961 ctccgaaata cttggttggc gtgtttcgta atcaacctaa ggaggatgtt ttggctctgg
     1021 tcaatgatta cggcattgat atcgtccaac tgcatggaga tgagtcgtgg caagaatacc
     1081 aagagttcct cggtttgcca gttattaaaa gactcgtatt tccaaaagac tgcaacatac
     1141 tactcagtgc agcttcacag aaacctcatt cgtttattcc cttgtttgat tcagaagcag
     1201 gtgggacagg tgaacttttg gattggaact cgatttctga ctgggttgga aggcaagaga
     1261 gccccgaaag cttacatttt atgttagctg gtggactgac gccagaaaat gttggtgatg
     1321 cgcttagatt aaatggcgtt attggtgttg atgtaagcgg aggtgtggag acaaatggtg
     1381 taaaagactc taacaaaata gcaaatttcg tcaaaaatgc taagaaatag gttattactg
     1441 agtagtattt atttaagtat tgtttgtgca cttgcctgat aacttcgtat agcatacatt
     1501 atacgaagtt atcccgggta ccgagctcga attcaacgaa gcatctgtgc ttcattttgt
     1561 agaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt
     1621 tacagaacag aaatgcaacg cgaaagcgct attttaccaa cgaagaatct gtgcttcatt
     1681 tttgtaaaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc
     1741 atttttacag aacagaaatg caacgcgaga gcgctatttt accaacaaag aatctatact
     1801 tcttttttgt tctacaaaaa tgcatcccga gagcgctatt tttctaacaa agcatcttag
     1861 attacttttt ttctcctttg tgcgctctat aatgcagtct cttgataact ttttgcactg
     1921 taggtccgtt aaggttagaa gaaggctact ttggtgtcta ttttctcttc cataaaaaaa
     1981 gcctgactcc acttcccgcg tttactgatt actagcgaag ctgcgggtgc attttttcaa
     2041 gataaaggca tccccgatta tattctatac cgatgtggat tgcgcatact ttgtgaacag
     2101 aaagtgatag cgttgatgat tcttcattgg tcagaaaatt atgaacggtt tcttctattt
     2161 tgtctctata tactacgtat aggaaatgtt tacattttcg tattgttttc gattcactct
     2221 atgaatagtt cttactacaa tttttttgtc taaagagtaa tactagagat aaacataaaa
     2281 aatgtagagg tcgagtttag atgcaagttc aaggagcgaa aggtggatgg gtaggttata
     2341 tagggatata gcacagagat atatagcaaa gagatacttt tgagcaatgt ttgtggaagc
     2401 ggtattcgca atattttagt agctcgttac agtccggtgc gtttttggtt ttttgaaagt
     2461 gcgtcttcag agcgcttttg gttttcaaaa gcgctctgaa gttcctatac tttctagaga
     2521 ataggaactt cggaatagga acttcaaagc gtttccgaaa acgagcgctt ccgaaaatgc
     2581 aacgcgagct gcgcacatac agctcactgt tcacgtcgca cctatatctg cgtgttgcct
     2641 gtatatatat atacatgaga agaacggcat agtgcgtgtt tatgcttaaa tgcgtactta
     2701 tatgcgtcta tttatgtagg atgaaaggta gtctagtacc tcctgtgata ttatcccatt
     2761 ccatgcgggg tatcgtatgc ttccttcagc actacccttt agctgttcta tatgctgcca
     2821 ctcctcaatt ggattagtct catccttcaa tgctatcatt tcctttgata ttggatcata
     2881 cgaattcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     2941 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     3001 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     3061 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     3121 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3181 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3241 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3301 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3361 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3421 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3481 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3541 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3601 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3661 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3721 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3781 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3841 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3901 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3961 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4021 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4081 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4141 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4201 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     4261 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4321 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4381 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4441 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4501 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4561 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4621 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4681 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4741 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4801 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4861 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4921 tcttcctttt tcaatattac cgcgaatcct tacatcacac ccaatccccc acaagtgatc
     4981 ccccacacac catagcttca aaatgtttct actccttttt tactcttcca gattttctcg
     5041 gactccgcgc atcgccgtac cacttcaaaa cacccaagca cagcatacta aatttcccct
     5101 ctttcttcct ctagggtgtc gttaattacc cgtactaaag gtttggaaaa gaaaaaagag
     5161 accgcctcgt ttctttttct tcgtcgaaaa aggcaataaa aatttttatc acgtttcttt
     5221 ttcttgaaaa tttttttttt tgattttttt ctctttcgat gacctcccat tgatatttaa
     5281 gttaataaac ggtcttcaat ttctcaagtt tcagtttcat ttttcttgtt ctattacaac
     5341 tttttttact tcttgctcat tagaaagaaa gcatagcaat ctaatctaag ttttaattac
     5401 aaaactagtg atatctgcgc actcgagtca tgtaattagt tatgtcacgc ttacattcac
     5461 gccctccccc cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg
     5521 tccctattta tttttttata gttatgttag tattaagaac gttatttata tttcaaattt
     5581 ttcttttttt tctgtacaga cgcgtgtacg catgtaacat tatactgaaa accttgcttg
     5641 agaaggtttt gggacgctcg aaggctttaa tttgcggcca atattattga agcatttatc
     5701 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5761 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     5821 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtc
//
