LOCUS       sequence-348140-        4462 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Xenopus oocyte expression vector containing YOR307C. Contains T7 RNA
            polymerase binding site, and adds polyA tail to YOR307C. 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 4462)
  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 4462)
  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: "Front 
            Microbiol."; date: "2024-04-12"; volume: "15:1376653. doi"; pages: "
            10.3389/fmicb.2024.1376653"
COMMENT     Sequence Label: pCfB8590
FEATURES             Location/Qualifiers
     source          1..4462
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     51..73
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     65..82
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     66..82
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     89..108
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        89..107
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           139..181
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     3'UTR           1677..1802
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     promoter        complement(1892..1910)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1893..1910)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1928..1944)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1928..1944)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1941..1963)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1952..1968
                     /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(1976..1982,1983..2000,2001..2006))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2021..2042
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2159..2176)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2330..2918)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2410..2429)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3089..3880,3881..3949))
                     /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     3712..3731
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3950..4054)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4122..4140
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4178..4200)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4300..4319
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
       61 acgttgtaaa acgacggcca gtgaattgta atacgactca ctatagggcg aattaattcg
      121 agctcggtac ccagcttgct tgttcttttt gcagaagctc agaataaacg ctcaactttg
      181 gcagatcaat tcccggggct gaggcttaat atgattcaaa cgcaaagtac agcgatcaaa
      241 cgacgtaatt ctgttcataa aaacctcttt gatccatcac tgtatcaaat accagaacca
      301 ccacgtgggg ggttccaaca ccaaaaaaaa gaatacagta aggagacatt cagtaatcaa
      361 gtctttggat atgatattac tagccttaaa aaaagattca cgcaactatt tcctagtaat
      421 atacaagggt acttacctga agttgaccta agaataacca ttatttgttc tatatggtac
      481 gtcacgtcat ctatttcaag taacctatct aaagccattt taagaacttt caaccatccc
      541 atagctctca cagaattgca atttcttgtt agtgctgttc tatgtgttgg atttgcttcc
      601 atcgtaaatt tattccgact acctcgtttg aagcatacga agttttcaaa ggcactcaat
      661 agtttccctg acggtatcct tcctgaatat ctcgatggca atttcaggag ttctatcctg
      721 cacaagtttt tagttccctc aaagctggtt ttgatgacca ctttccctat gggaatattc
      781 caatttattg gtcatatcac atcgcacaag gcggtatcta tgataccagt atcactagtg
      841 cattccgtga aggcattatc cccaataata acagttggct actataaatt tttcgaacat
      901 cgttattaca attccatgac ttattatacc ttgttacttt taatttttgg cgttatgact
      961 acttgctggt caacacatgg cagtaagagg gcttcagata acaaaagcgg atcttcatta
     1021 atcgggttgc tttttgcctt tatttccatg ataatatttg tagcacagaa tatttttgca
     1081 aagaatattt taaccatcag aaggaaggta ggaatactgc cgtcttcttc tacggatgac
     1141 gtcacgtcga aggaagggca accgagtcta gacaaaacaa gattttctcc attgcaagtg
     1201 gataagatta ccatattgtt ctactgctcc tgcattgggt tttctttaac cctattacct
     1261 tttttaaccg gcgaattaat gcatggcggt agcgttatca acgatttaac gctagaaaca
     1321 gtagcccttg tagcgattca tggaatagcc cattttttcc aagcaatgct tgctttccag
     1381 ttgatcggtt tactatcttc cattaattat tcggtagcaa acatcatgaa gaggattgtt
     1441 gttatatccg tggcactctt ttgggaaaca aagttaaatt tttttcaggt gtttggtgtt
     1501 atcttgacaa ttgccggatt gtacggttat gacaaatggg ggctttccaa aaaagatgga
     1561 cgtcaggcat gaattaaacc tcagcgaatt cgtgagaaag cttgggaata ggcctttcag
     1621 atctgcccga gaccatggat gaagtatcta gaaatagctt gatctggtta ccactaaacc
     1681 agcctcaaga acacccgaat ggagtctcta agctacataa taccaactta cactttacaa
     1741 aatgttgtcc cccaaaatgt agccattcgt atctgctcct aataaaaara aagtttcttc
     1801 acattctaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa amcccccccc cccccccctg
     1861 caggcggccg cctcgaggct agcttgagta ttctatagtg tcacctaaat agcttggcgt
     1921 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     1981 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     2041 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     2101 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     2161 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     2221 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     2281 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     2341 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     2401 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     2461 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2521 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2581 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2641 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2701 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2761 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     2821 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     2881 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     2941 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     3001 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     3061 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     3121 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     3181 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     3241 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     3301 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     3361 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     3421 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     3481 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     3541 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     3601 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     3661 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     3721 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     3781 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     3841 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     3901 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     3961 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     4021 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     4081 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     4141 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     4201 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     4261 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     4321 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     4381 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     4441 cttcgctatt acgccagctg gc
//
