LOCUS       sequence-260078-        4923 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4923)
  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 4923)
  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
FEATURES             Location/Qualifiers
     source          1..4923
                     /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(3048..3147)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3164..3183)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3165..3183)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3188..3204)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3188..3204)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3306..3325)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3317..4126
                     /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(3384..3403)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4273..4861
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4762..4781
                     /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 ggccacgacc cagctttctt
     3061 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     3121 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta
     3181 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     3241 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     3301 taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc
     3361 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
     3421 tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg
     3481 caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga
     3541 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     3601 caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     3661 tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     3721 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     3781 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     3841 agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg
     3901 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     3961 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     4021 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     4081 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa
     4141 ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc
     4201 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     4261 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     4321 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     4381 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     4441 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4501 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4561 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4621 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4681 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4741 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4801 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4861 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4921 gtt
//
