LOCUS       sequence-348277-        6464 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Xenopus oocyte expression vector containing YBR235W. Contains T7 RNA
            polymerase binding site, and adds polyA tail to YBR235W. 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 6464)
  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 6464)
  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: pCfB8153
FEATURES             Location/Qualifiers
     source          1..6464
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     173..191
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(229..251)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     351..370
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     564..586
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     578..595
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     579..595
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     602..621
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        602..620
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           652..694
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     3'UTR           4191..4316
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     promoter        complement(4407..4425)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4408..4425)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4443..4459)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4443..4459)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4456..4478)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4467..4483
                     /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(4491..4497,4498..4515,4516..4521))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4536..4557
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4674..4691)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4845..5433)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4925..4944)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5604..6395,6396..6464))
                     /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     6227..6246
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
      181 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
      241 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
      301 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
      361 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
      421 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
      481 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
      541 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt
      601 gtaatacgac tcactatagg gcgaattaat tcgagctcgg tacccagctt gcttgttctt
      661 tttgcagaag ctcagaataa acgctcaact ttggcagatc aattcccggg gctgaggctt
      721 aatatggtta gtaggtttta tcagattcca ggtacgcatc ggccatcgtc ggcaatatct
      781 tcttcaaatg aatcttcttc actgttgtct gcaaggagaa taagtcaaac atatttcaat
      841 tatcaggcga ctcctgaatg ccaaaaagtt tcctctaagt atgatcctga taacccaaac
      901 aaagataagt tgggaacata cgatggggta tttgtgccta ctgctttaaa cgtattgtct
      961 atccttatgt ttcttcgttt tggcttcatt ttgggtcagt taggtattat atgcaccatc
     1021 ggtcttctgc tattgagtta cactattaat cttctcacca cgttaagtat atctgctata
     1081 tctacaaacg ggactgttag gggcgggggt gcttattata tgatttcaag aagtttagga
     1141 cctgaatttg gtggatccat tgggcttgtc ttcttcttag ggcaggtgtt caatgcaggt
     1201 atgaacgcag tgggtattat cgaaccttta ctctataact tgggttattc tgctcaaggc
     1261 gagccacctg cagctttggg agagctacta ccaagagggc attggcacga atttacatat
     1321 gccacggtca ttcttttctt atgtttttct gtggcttttg ttggctcgca aacagtgtca
     1381 agagcaggga atattctttt tttggtactg gcagcttcta tattttccat tccactttct
     1441 gcattgatta gatcgccatt cactgaaggc ggtatcagtt atacaggtcc ttcatggcaa
     1501 acttttcatg ataacctgct ccctcactta acaaaaggtg cagcaggttc gttactcaaa
     1561 ggtaaggaaa cattcaatga tttatttggc gttttttttc ctgctacagc tggtatattc
     1621 gcgggcgcag gaatgtcaag tgaattgaga aagccctcta aatcaattcc taagggtacc
     1681 ttatggggtt tactattcac ttttatctgt tatgccgttg ttgtcttctc catgggttgt
     1741 tccattccca gaagatcgtt atatgatgaa gtacaaatta tacagacgat cagttccgtt
     1801 caatgggtta tattcatggg tgaaatggct acttctcttt tctccatcat agtggggatg
     1861 ctcggtgctg cttatgtatt agaagcaatt gcaaaggata acattattcc tggtttggag
     1921 atttttgctc attcaccgtt atattcattg atttttactt ggattctaac acagctatgt
     1981 ttgttttcag acgtcaataa aatcgccaca tttatcacaa tgacattctt aatgacgttt
     2041 gtggtcatga atttggcgtg cttcctatta ggtatttcct ctgctcccaa tttcaggccc
     2101 tctttcaaat actttaacag gtatactaca gcaattggcg cattactttc tgttgttgcg
     2161 atgttaatag ttgatggtat ctcagcatct gtcttgtttt tggctatgat tttgctattt
     2221 ttattcattc attatttttc accaccgaag tcttggggtg atgtgtctca aagtttaata
     2281 tatcatcaag tgagaaaata tttgcttcgt ctacgtcaag acaatattaa atactggagg
     2341 cctcaaatat tgctgtttgt agataatcca aggacgagct ggaacttaat aaggttttgc
     2401 aatcatttga aaaaaggtgg tttgtatatt ttaggccatg ttgcagtaac tgcggatttt
     2461 ccaaagcaac taaatgaatt aaaaactcag cagaaagctt ggatgaagat cagagacatg
     2521 gctgcaataa aggcttttgt ccaagttggg actggtccct cccttatatg gggaattagg
     2581 aatgttttta ttggttccgg gcttggaggt atgaaaccaa acattactgt tgtcggtttt
     2641 tttgacctcg aaagttatag aaagcacatc ccacaaagtc gcagccaaaa taaccttcag
     2701 aaacaggtgg aaataaaagc tacagtgcca cgtagtacat gctctgatgt caaaatcaat
     2761 gttccattgc caacggatga gtgtaaaaat gaaaccaaag ttaatgtaca acaatgggtt
     2821 caaattgtag aagatctatc actgatgcaa tccaatattg ccatagcgca tggattcaaa
     2881 aatttagaga taccgaacaa aagggaccgc tgctttccaa aaaaaactat agatctttat
     2941 cccatccaaa tgtgcggaaa agttgaggca aagggtgatc agcctgcggc tatcactact
     3001 aactttgata cgtatacact gattctacaa ttagctgcca ttttggttac tgttcctgaa
     3061 tggaagcgca cacattcgct tcgagttatt ttatttgtgg aacaagaata tcatagaacg
     3121 aacgaaactc aacgaatgaa aaagctatta caggttctaa gaattgacgc agaggttttg
     3181 gtagtatctt tagatcaatt cagagtatac aacacaattg tcaagggaga cccaatcgtg
     3241 tttgattatg tcaactcgaa attggccgat aatgaatggt ggaaagattt ggttgaagct
     3301 cgtgatacat taaaaccgaa gcggaggttt tccactattg aacctcaaac gatagcaaag
     3361 caatttacac aatcaagaaa atatacgtct ggggttcaga aattgggtgt ctcatttaca
     3421 atgaatacga gaatgccaac taaccgcatt gataccccat gcgagagtga ggactccgat
     3481 ttggatacag atcttacgtc aattcgtgat gccttctcgg catcgactaa tatttcagta
     3541 ggtaaggact taacgaccaa atcgaagacc ggttccgaca gaacaaattt actcgtaaag
     3601 attttacaaa gcgatgtgtc aacacagtcg ttaagacccg ttttctcgag taatactttg
     3661 ccgaggacga gagtcgtgga agatggcacc ggtgaacagc cgacactgat cccaattgcc
     3721 gaacctgacc tttcaaatgg gaacggtact gggagcggta ttggcaacgg caacaaatta
     3781 aagaaaccgg ttcttcccga attatcacct tgttgttcca aagatagttt agtcacagcg
     3841 atgcaaaatt taggtttcaa tgatcttccc agcactgctc aacatttggt tctgaatgat
     3901 gtgatgacac aaatgtcaaa gagttcggat ttaatttttt caaccctgcc agttccagct
     3961 cttggaacac atgaagatca cgatgcaagt ttacaatatg tcgaggactt ggatatttgg
     4021 ttggagggct tacctccatg catgttaatc aattcgcaaa ccatgacagt aactactgca
     4081 ttatgaatta aacctcagcg aattcgtgag aaagcttggg aataggcctt tcagatctgc
     4141 ccgagaccat ggatgaagta tctagaaata gcttgatctg gttaccacta aaccagcctc
     4201 aagaacaccc gaatggagtc tctaagctac ataataccaa cttacacttt acaaaatgtt
     4261 gtcccccaaa atgtagccat tcgtatctgc tcctaawaaa aaraaagttt cttcacattc
     4321 taaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaamcccc cccccccccc ccctgcaggc
     4381 ggccgcctcg aggctagctt gagtattcta tagtgtcacc taaatagctt ggcgtaatca
     4441 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     4501 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     4561 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     4621 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     4681 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     4741 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     4801 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4861 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4921 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4981 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5041 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5101 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5161 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5221 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5281 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5341 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5401 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5461 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5521 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5581 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5641 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5701 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     5761 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     5821 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     5881 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     5941 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6001 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6061 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6121 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6181 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6241 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6301 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6361 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6421 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcat
//
