LOCUS       sequence-12704-e        5010 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5010)
  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. 2010 Sep 10. ():.
  PUBMED    20936606
REFERENCE   2  (bases 1 to 5010)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Yeast. 2010
            Sep 10. ():."
COMMENT     Sequence Label: pXP316
FEATURES             Location/Qualifiers
     source          1..5010
                     /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)."
     primer_bind     complement(2042..2058)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2042..2058)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2055..2077)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2066..2082
                     /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(2090..2096,2097..2114,2115..2120))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2135..2156
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2273..2290)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2444..3032)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2524..2543)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3203..3994,3995..4063))
                     /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     3826..3845
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        4122..4535
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      4573..4820
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4589..4607)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     4983..5001
                     /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 attcatcacg tgctataaaa ataattataa
     1561 tttaaatttt ttaatataaa tatataaatt aaaaatagaa agtaaaaaaa gaaattaaag
     1621 aaaaaatagt ttttgttttc cgaagatgta aaagactcta gggggatcgc caacaaatac
     1681 taccttttat cttgctcttc ctgctctcag gtattaatgc cgaattgttt catcttgtct
     1741 gtgtagaaga ccacacacga aaatcctgtg attttacatt ttacttatcg ttaatcgaat
     1801 gtatatctat ttaatctgct tttcttgtct aataaatata tatgtaaagt acgctttttg
     1861 ttgaaatttt ttaaaccttt gtttattttt ttttcttcat tccgtaactc ttctaccttc
     1921 tttatttact ttctaaaatc caaatacaaa acataaaaat aaataaacac agagtaaatt
     1981 cccaaattat tccatcatta aaagatacga ggcgcgtgta agttacgaat tcgtaatcat
     2041 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     2101 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     2161 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     2221 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     2281 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     2341 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     2401 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     2461 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     2521 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     2581 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     2641 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     2701 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     2761 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     2821 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     2881 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     2941 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     3001 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     3061 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     3121 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     3181 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     3241 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     3301 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     3361 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     3421 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     3481 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     3541 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     3601 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     3661 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     3721 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     3781 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     3841 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     3901 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     3961 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     4021 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     4081 attaccgcga atccttacat cacacccaat cccccacaag tgatccccca cacaccatag
     4141 cttcaaaatg tttctactcc ttttttactc ttccagattt tctcggactc cgcgcatcgc
     4201 cgtaccactt caaaacaccc aagcacagca tactaaattt cccctctttc ttcctctagg
     4261 gtgtcgttaa ttacccgtac taaaggtttg gaaaagaaaa aagagaccgc ctcgtttctt
     4321 tttcttcgtc gaaaaaggca ataaaaattt ttatcacgtt tctttttctt gaaaattttt
     4381 ttttttgatt tttttctctt tcgatgacct cccattgata tttaagttaa taaacggtct
     4441 tcaatttctc aagtttcagt ttcatttttc ttgttctatt acaacttttt ttacttcttg
     4501 ctcattagaa agaaagcata gcaatctaat ctaagtttta attacaaaac tagtgatatc
     4561 tgcgcactcg agtcatgtaa ttagttatgt cacgcttaca ttcacgccct ccccccacat
     4621 ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt
     4681 ttatagttat gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt
     4741 acagacgcgt gtacgcatgt aacattatac tgaaaacctt gcttgagaag gttttgggac
     4801 gctcgaaggc tttaatttgc ggccaatatt attgaagcat ttatcagggt tattgtctca
     4861 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     4921 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     4981 aaaataggcg tatcacgagg ccctttcgtc
//
