LOCUS       sequence-348279-        4827 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Xenopus oocyte expression vector containing YBL020W. Contains T7 RNA
            polymerase binding site, and adds polyA tail to YBL020W. 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 4827)
  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 4827)
  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: pCfB8151
FEATURES             Location/Qualifiers
     source          1..4827
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     24..43
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        24..42
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           74..116
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     3'UTR           1975..2100
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     promoter        complement(2192..2210)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2193..2210)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2228..2244)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2228..2244)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2241..2263)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2252..2268
                     /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(2276..2282,2283..2300,2301..2306))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2321..2342
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2459..2476)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2630..3218)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2710..2729)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3389..4180,4181..4249))
                     /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     4012..4031
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4250..4354)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4422..4440
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4478..4500)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4600..4619
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(4813..4827,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4827,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttgtaatacg actcactata gggcgaatta attcgagctc
       61 ggtacccagc ttgcttgttc tttttgcaga agctcagaat aaacgctcaa ctttggcaga
      121 tcaattcccg gggctgaggc ttaatatggc gaaaaaaaac tcacaattgc cctctactag
      181 cgagcagatc ttggaaaggt ccacaacagg agctaccttt ctcatgatgg gccaactttt
      241 caccaaactg gtaacgttca tactaaataa tttgttgatc aggtttctgt cgcccagaat
      301 tttcggtatc acggcctttc tagaatttat acagggcaca gtgttatttt ttagcagaga
      361 tgcgattcgt ctgtcgacgt tgagaatctc agactccggt aatggaataa tcgatgatga
      421 cgacgaggag gagtaccagg aaactcatta caagtctaaa gttttgcaaa ccgcagtcaa
      481 ttttgcttac attccgtttt ggatcgggtt tccactgtcc attggtctta tcgcctggca
      541 gtacagaaac atcaacgcgt atttcatcac tcttccattc ttcaggtggt cgatttttct
      601 tatctggctg agtatcatcg tggagctgtt aagcgagcca ttcttcatcg tcaaccagtt
      661 tatgttgaac tatgccgcaa ggtcaagatt tgaaagcatc gcggtgacta caggatgtat
      721 tgtcaatttt atagttgttt atgccgttca gcaatcccgc tacccaatgg gggttgtcac
      781 atcggacatt gacaaagaag gcatcgccat attggcattt gccttgggaa agttagcaca
      841 ttcgatcacc ctgctagcat gttactactg ggactatctc aagaatttca aaccaaagaa
      901 attgttcagt accaggctaa cgaagataaa aacgcgtgaa aataacgaat tgaagaaagg
      961 ctacccaaag agcacatctt attttttcca aaacgacatt ttacagcact tcaaaaaagt
     1021 ttattttcaa ctatgtttta agcatttgtt gacagagggt gataagttga ttatcaattc
     1081 tttatgtact gtggaagaac aaggcattta cgctctattg tcgaactatg gatcgctact
     1141 aacaagatta ttatttgcgc cgatcgaaga atctctgcgg ttatttttgg cccgtttatt
     1201 atcctcgcat aaccctaaaa atttaaaact atctattgaa gtcctggtga atttaacaag
     1261 gttttacata tacttatcgt taatgatcat tgtatttggg cctgccaatt catccttttt
     1321 attgcagttc ttgattggct cgaaatggtc cactacttcc gttttggaca ctataagagt
     1381 ctactgcttt tacatcccat ttttatcgct taatggtatt tttgaagctt ttttccagag
     1441 tgtagccact ggtgaccaaa ttttgaaaca ttcatatttt atgatggcct tttctggtat
     1501 tttcctgctc aattcctggc ttcttattga aaaactcaaa ctatcaatcg aaggcttgat
     1561 attgagtaac atcattaaca tggtgttgag aatattgtat tgtggagttt tcttgaataa
     1621 atttcatagg gaactgttta cagattcctc ttttttcttc aattttaagg atttcaaaac
     1681 agttattatt gctggctcaa cgatctgtct acttgactgg tggtttattg ggtacgttaa
     1741 aaatttacaa caatttgttg ttaacgtatt attcgcaatg ggattgttag cgttaatttt
     1801 ggtcaaggag cgccaaacca tacaatcttt tattaacaag agggcggttt ccaattctaa
     1861 agatgtatga attaaacctc agcgaattcg tgagaaagct tgggaatagg cctttcagat
     1921 ctgcccgaga ccatggatga agtatctaga aatagcttga tctggttacc actaaaccag
     1981 cctcaagaac acccgaatgg agtctctaag ctacataata ccaacttaca ctttacaaaa
     2041 tgttgtcccc caaaatgtag ccattcgtat ctgctcctaa taaaaaraaa gtttcttcac
     2101 attctaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaam cccccccccc cccccccctg
     2161 caggcggccg cctcgaggct agcttgagta ttctatagtg tcacctaaat agcttggcgt
     2221 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     2281 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     2341 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     2401 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     2461 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     2521 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     2581 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     2641 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     2701 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     2761 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2821 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2881 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2941 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     3001 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     3061 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     3121 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     3181 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     3241 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     3301 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     3361 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     3421 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     3481 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     3541 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     3601 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     3661 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     3721 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     3781 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     3841 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     3901 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     3961 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     4021 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     4081 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     4141 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     4201 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     4261 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     4321 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     4381 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     4441 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     4501 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     4561 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     4621 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     4681 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     4741 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     4801 cgccagggtt ttcccagtca cgacgtt
//
