LOCUS       sequence-444348-        4926 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors. Contains a STOP codon at the end of 
            the codon-optimized ORF sequence. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4926)
  AUTHORS   Superti-Furga G, Lackner D, Wiedmer T, Ingles-Prieto A, Barbosa B, 
            Girardi E, Goldmann U, Gurtl B, Klavins K, Klimek C, Lindinger S, 
            Lineiro-Retes E, Muller AC, Onstein S, Redinger G, Reil D, Sedlyarov
            V, Wolf G, Crawford M, Everley R, ... Steppan CM
  TITLE     The RESOLUTE consortium: unlocking SLC transporters for drug 
            discovery.
  JOURNAL   Nat Rev Drug Discov. 2020 Jul;19(7):429-430. doi: 
            10.1038/d41573-020-00056-6.
  PUBMED    32265506
REFERENCE   2  (bases 1 to 4926)
  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"; doi: "10"; journalName: 
            "Nat Rev Drug Discov"; date: "2020-07"; volume: "19"; issue: "7"; 
            pages: "429-430"
COMMENT     Sequence Label: pDONR221-MFSD6_STOP
FEATURES             Location/Qualifiers
     source          1..4926
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3051..3150)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3167..3186)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3168..3186)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3191..3207)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3191..3207)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3309..3328)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3320..4129
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3387..3406)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4276..4864
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4765..4784
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacacattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc accatggccg acgataaggt ggccatcctg accgacgatg aggaggagca
      721 gaagcggaag tacgtgctgg ccgacccttt caacggcatc tccagagagc cagagccccc
      781 ttctaatgag acacccagct ccaccgagac atccgccatc cccgaggagg agatcgactg
      841 gatcgagaag cactgcgtga agatcaacaa tgatctgctg atctctaagg tgttttactt
      901 ctttttctac tccgcctatg gctctctgta tcctctgctg ccagtgtact ataagcagct
      961 gggcatgagc ccatcccagt ctggactgct ggtgggcatc cggtacttca tcgagttttg
     1021 ttctgcccct ttctggggag tggtggcaga tcggttcaag aagggcaaga tcgtgctgct
     1081 gttcagcctg ctgtgctggg tgctgttcaa cctgggcatc ggctttgtga agcctgccac
     1141 cctgagatgc gtgcccaaga tccgccctac cacacaccca acaaacgcca gccaccagct
     1201 gaccatcctg ccaacaaatt ctagcttcac cagctttctg acaatctccc ccaagatgcg
     1261 ggagaagaga aacctgctgg agacccggct gaacgtgagc gacaccgtga cactgccaac
     1321 cgcacctaac atgaatagcg agcccacact gcagcctcag accggagaga tcacaaaccg
     1381 catgatggat ctgaccctga attcctctac cgccacacca gtgtctcccg gcagcgtgac
     1441 aaaggagacc acaaccgtga tcgtgacaac cacaaagtcc ctgccctctg accaggtcat
     1501 gctggtgtac gatcagcagg aggtggaggc catcttcctg gtcatcctgg tggtggtcat
     1561 catcggcgag tttttctccg ccagctccgt gaccatcgtg gacaccgtga cactgcagta
     1621 cctgggcaag caccgggata gatatggact gcagaggatg tggggaagcc tgggatgggg
     1681 actggcaatg ctgtccgtgg gaatcggcat cgactacacc cacatcgagg tgctgatcga
     1741 tggcaagggc tgcaagccac ccgagtaccg caattatcag atcgtgttca tcgtgtttgg
     1801 cgtgctgatg accatggccc tgatcgtggc cacacagttt aggttccgct ataaccactt
     1861 caagaatgac gattccaagg gcaaggaggt ggagatccct caggtggagc ggaacaattc
     1921 taccgagtct agcgaggaga caccaaccac aaccagccac tcccaggcct ttaacttctg
     1981 ggacctgatc aagctgctgt gctctgtgca gtacggcagc gtgctgttcg tggcctggtt
     2041 catgggcttt ggctacggct tcgtgttcac cttcctgtat tggcacctgg aggatctgaa
     2101 tggcacaacc acactgtttg gcgtgtgcag cgtgctgtcc cacgtgtctg agctgaccgc
     2161 ctacttcttc agccacaagc tgatcgagct gatcggccac atcagggtgc tgtacatcgg
     2221 cctggcctgt aacacagccc gctacatcta tatctcctat ctggagaatg cctggaccgt
     2281 gctgcctatg gaggtgctgc agggagtgac acacgcagca atctgggcag catgcatcag
     2341 ctacctgtcc gccgcagtgc ctccagagct gaggaccagc gcccagggca tcctgcaggg
     2401 cctgcacctg ggactgggaa ggggatgcgg agccatgatc ggcggcgtgc tggtgaacta
     2461 tttcggagca gcagccacct ttcggggaat cggcatggcc tgtctggtca tcctgctgct
     2521 gttcgccctg atccagtggc tggccgtgcc tgacgaggag gaggataaga caatgctggc
     2581 cgagagaatc ccagtgccct cctctcctgt gccaatcgcc accatcgacc tggtgcagca
     2641 gcagacagag gatgtgatgc cacggatcga gcccagactg ccccctaaga agaccaagca
     2701 ccaggaggag caggaggacg tgaacaagcc tgcctggggc gtgagctcct ctccatgggt
     2761 gaccttcgtg tacgccctgt atcagatcaa ggagatgatg cagctgacca gggataacag
     2821 ggcaagcgag atccagccac tgcagggcac aaacgagaat cgggagaatt cccctgcagg
     2881 aagagcacag ccagtgcctt gcgagaccca ctctgaccca agcaggaatc agccaagccc
     2941 agatgcagca gcatcccaga cccagacatc tcctgcccac ccaagcgtgg acccctgtac
     3001 cgaggagtcc gaggagcagc aggcacagct ggccgccggc ggccactaag acccagcttt
     3061 cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt
     3121 cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc
     3181 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     3241 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     3301 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     3361 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     3421 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     3481 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     3541 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     3601 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     3661 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     3721 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     3781 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     3841 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     3901 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     3961 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     4021 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     4081 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     4141 taattggttg taacactggc agagcattac gctgacttga cgggacggcg caagctcatg
     4201 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     4261 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4321 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     4381 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     4441 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4501 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4561 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4621 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4681 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4741 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4801 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4861 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4921 catgtt
//
