LOCUS       sequence-348538-        4282 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Xenopus oocyte expression vector containing YML018C. Contains T7 RNA
            polymerase binding site, and adds polyA tail to YML018C. 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 4282)
  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 4282)
  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: pCfB8586
FEATURES             Location/Qualifiers
     source          1..4282
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           51..93
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     3'UTR           1409..1534
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     promoter        complement(1624..1642)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1625..1642)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1660..1676)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1660..1676)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1673..1695)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1684..1700
                     /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(1708..1714,1715..1732,1733..1738))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1753..1774
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1891..1908)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2062..2650)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2142..2161)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2821..3612,3613..3681))
                     /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     3444..3463
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3682..3786)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3854..3872
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3910..3932)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4032..4051
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4245..4267
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4259..4276
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4260..4276
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactataggg cgaattaatt cgagctcggt acccagcttg cttgttcttt
       61 ttgcagaagc tcagaataaa cgctcaactt tggcagatca attcccgggg ctgaggctta
      121 atatggtgtc gaaggatcaa acgtccttta ataagaggtg gaccttgggt cttctgatgt
      181 tgggtctagt cattattttg tgggttttat cgtccttcct gataaatttg atcttcgagg
      241 atgactcgta tagaaaacca ttctttatca cgtacaccaa tactgccgcc tttatctttt
      301 atctttttcc aactgcaaag gccgttgtgg tgaattacaa ggacacaggc cgtgcaaacg
      361 ttcaccgcga actaattatg gaggaagaag gtactggtag tgatagcaat cgctcggttg
      421 atatgactag cccactgctt acaaaccttg aagcaggcac gcatgccaac caaaagaaga
      481 ggctaacgtt atatgaaaca atcaagttaa gtgctgagtt ctgcatactt tggttcacag
      541 ctaaccttgt gaccaatgcc tcgctggcat tcacatcggt ggcatcgcag acaatcctct
      601 ccactacttc ttcctttttc accttgttca taggtgctat ttgtcatgtt gaatcgttga
      661 gcaagtccaa agtactcggg tcgttcattt cctttgtagg cataatcatg gtcaccaaat
      721 cagattctca tcagcgctac caacgccata tcgcggacgt tagtggtgat gataacgatg
      781 ccgtgcaagt tctgataggc aacttgttgg cacttgcagg tgctgtactt tacggtgtct
      841 acagtacttt attgaagaga gaagtcggag atgagactcg tgttaacatg aagatttttt
      901 ttggctttgt tggtctcttt aatttacttt ttctatggcc atcactgatt gtattggact
      961 tttttggctg ggaaccattc tctctgccca aagatcctaa agttgtggta ataatcttcg
     1021 tcaattgtct catcacattt gttagcgact tttgttgggc aaaggctatg ttactaacga
     1081 gtcctttaac cgtcaccgta ggattgagca taacgatacc cctagcaatg tttggtgatg
     1141 taatctttaa gcacaaaacc atgtctgcgc tctatttatt tggtgctacc ctcatcttag
     1201 gctcattttt catcattaac aaaagctctg aggaggaaca ttttgagaac tcaataacgg
     1261 ctagcaatta cgaatcagtg gaggtgccag cagcaaacaa ctgaattaaa cctcagcgaa
     1321 ttcgtgagaa agcttgggaa taggcctttc agatctgccc gagaccatgg atgaagtatc
     1381 tagaaatagc ttgatctggt taccactaaa ccagcctcaa gaacacccga atggagtctc
     1441 taagctacat aataccaact tacactttac aaaatgttgt cccccaaaat gtagccattc
     1501 gtatctgctc ctaataaaaa gaaagtttct tcacattcta aaaaaaaaaa aaaaaaaaaa
     1561 aaaaaaaaaa aaamcccccc cccccccccc tgcaggcggc cgcctcgagg ctagcttgag
     1621 tattctatag tgtcacctaa atagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     1681 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     1741 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     1801 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     1861 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     1921 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     1981 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2041 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     2101 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2161 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     2221 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     2281 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     2341 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     2401 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     2461 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     2521 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     2581 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     2641 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     2701 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     2761 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     2821 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     2881 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     2941 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     3001 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     3061 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     3121 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     3181 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     3241 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     3301 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     3361 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     3421 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     3481 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     3541 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     3601 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     3661 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     3721 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     3781 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     3841 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     3901 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     3961 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     4021 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     4081 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg
     4141 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     4201 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     4261 taaaacgacg gccagtgaat tg
//
