LOCUS       sequence-363153-        4384 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Xenopus oocyte expression vector containing YBR287W. Contains T7 RNA
            polymerase binding site, and adds polyA tail to YBR287W. Used for 
            generation of mRNA, for injection into xenopus laevis oocytes..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4384)
  AUTHORS   Moller-Hansen I, Saez-Saez J, van der Hoek SA, Dyekjaer JD, 
            Christensen HB, Wright Muelas M, O'Hagan S, Kell DB, Borodina I
  TITLE     Deorphanizing solute carriers in Saccharomyces cerevisiae for 
            secondary uptake of xenobiotic compounds.
  JOURNAL   Front Microbiol. 2024 Apr 12;15:1376653. doi: 
            10.3389/fmicb.2024.1376653.
  PUBMED    38680917
REFERENCE   2  (bases 1 to 4384)
  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"; journalName: "Front 
            Microbiol."; date: "2024-04-12"; volume: "15:1376653. doi"; pages: "
            10.3389/fmicb.2024.1376653"
COMMENT     Sequence Label: pCfB8154
FEATURES             Location/Qualifiers
     source          1..4384
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     5'UTR           1..43
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     3'UTR           1461..1586
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     promoter        complement(1676..1694)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1677..1694)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1712..1728)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1712..1728)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1725..1747)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1736..1752
                     /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(1760..1766,1767..1784,1785..1790))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1805..1826
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1943..1960)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2114..2702)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2194..2213)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2873..3664,3665..3733))
                     /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     3496..3515
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3734..3838)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3906..3924
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3962..3984)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4084..4103
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4297..4319
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4311..4328
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4312..4328
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4335..4354
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4335..4353
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cttgttcttt ttgcagaagc tcagaataaa cgctcaactt tggcagatca attcccgggg
       61 ctgaggctta atatggttga aacatttagt tttgctcatt tggcttattt ggtctttgag
      121 tccgttttac aggtagtcat tatcgcactc gctggttttt ggagcgctag ttctggtctg
      181 ctgcccaaac aaagtcaaaa aatcatttct ctgttgaacg tggacctttt tacaccatgt
      241 cttattttta gtaagttggc caaatccctg tccatggcca aaatctttga aatagccatc
      301 attcctattt tttttggttt gactactggt atttcattta tttccggaaa gataatgagc
      361 cgtatcctag acttggataa agatgaaact aattttgtgg tggcaaactc tgtctttggt
      421 aacagtaact ctctacctgt gtccttgaca ttatctttgg cttacactct gcccaactta
      481 acttgggacc aaattcctaa tgataacaga gataacgttg catcaagggg tattttatac
      541 ttgctgatct ttcaacaaat cggacaaatg ctgaggtgga gctggggtta caataagcta
      601 atgaagtggt ctggagagaa cactcagcat atgccacctt ctcaagtgca atctttgttg
      661 gaaaggacgc ccaacattga taacgaggaa cttgtcaatg aggaacaaga ggaacaagaa
      721 ttgcttgaag aagaaaataa cagaatgaac tcatcttttt tgagttcgag ttccatcggt
      781 gataagattt ggcaaaaatc atgcactgta tttgagagaa tcagagcgaa ccttaatcct
      841 ccgttatatt ccatgatttt tgctgttgtt gtggcagcaa ttggtccgtt gcagagagag
      901 ctatttatgg aagatgggtt tatcaataac acatttgccg aggctgttac gcagctgggt
      961 tcagtttcta tcccactgat cctggttgtc ctcggttcta acttatatcc ttctgcggaa
     1021 gtcttcccta agacggttca ccatagcaag ttattgatcg gctccatcat cggaagaatg
     1081 attttacctt cttgcttcct tttgcctatt attgctattg ctgttaagta cataaacgtg
     1141 agtattttgg acgatccaat tttccttgtt gtcggttttc ttttgactgt ttcccctcct
     1201 gccattcaat taactcaaat cactcaactg aatgaatttt tcgaggcaga aatggccgat
     1261 attctatttt gggggtacgc tgtcttaagt ttgcctgtaa gtattattgt tgtctcaggt
     1321 gcaatttacg ttttacagtg ggctaaccca acttgaatta aacctcagcg aattcgtgag
     1381 aaagcttggg aataggcctt tcagatctgc ccgagaccat ggatgaagta tctagaaata
     1441 gcttgatctg gttaccacta aaccagcctc aagaacaccc gaatggagtc tctaagctac
     1501 ataataccaa cttacacttt acaaaatgtt gtcccccaaa atgtagccat tcgtatctgc
     1561 tcctaataaa aagaaagttt cttcacattc taaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     1621 aaaaamcccc cccccccccc cctgcaggcg gccgcctcga ggctagcttg agtattctat
     1681 agtgtcacct aaatagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     1741 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     1801 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     1861 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     1921 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     1981 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2041 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2101 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2161 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2221 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2281 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     2341 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2401 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     2461 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2521 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     2581 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2641 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2701 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     2761 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     2821 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     2881 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     2941 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     3001 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     3061 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     3121 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     3181 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     3241 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     3301 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     3361 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     3421 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     3481 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     3541 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     3601 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     3661 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     3721 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     3781 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     3841 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     3901 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     3961 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     4021 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     4081 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     4141 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt
     4201 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     4261 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     4321 cggccagtga attgtaatac gactcactat agggcgaatt aattcgagct cggtacccag
     4381 cttg
//
