LOCUS       sequence-259216-        3972 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 3972)
  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 3972)
  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_MFSD9
FEATURES             Location/Qualifiers
     source          1..3972
                     /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(2097..2196)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2213..2232)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2214..2232)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2237..2253)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2237..2253)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2355..2374)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2366..3175
                     /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(2433..2452)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3322..3910
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3811..3830
                     /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 accatggagc tgggaggaca ctgggacatg aactccgccc ctaggctggt
      721 gtctgagacc gcagagagaa agcaggagca gaagaccggc acagaggccg aggcagcaga
      781 cagcggagca gtgggagcac ggagatttct gctgtgcctg tacctggtgg gctttctgga
      841 tctgttcggc gtgtccatgg tggtgccact gctgtctctg cacgtgaaga gcctgggagc
      901 atccccaaca gtggcaggaa tcgtgggaag ctcctacgga atcctgcagc tgttctctag
      961 caccctggtg ggctgctggt ctgatgtggt gggcaggcgc tcctctctgc tggcatgtat
     1021 cctgctgagc gccctgggat atctgctgct gggagcagcc acaaacgtgt tcctgtttgt
     1081 gctggcaagg gtgccagcag gcatctttaa gcacaccctg tctatcagcc gggccctgct
     1141 gagcgacgtg gtgccagaga aggagagacc cctggtcatc ggccacttta acacagcatc
     1201 cggagtgggc ttcatcctgg gaccagtggt gggaggatac ctgaccgagc tggaggatgg
     1261 cttctatctg acagccttca tctgttttct ggtgttcatc ctgaatgccg gcctggtgtg
     1321 gttctttcct tggagggagg caaagccagg atccaccgag aagggactgc cactgaggaa
     1381 gacacacgtg ctgctgggcc gctctcacga caccgtgcag gaggcagcca caagcaggag
     1441 agcaagggcc tccaagaaga ccgcacagcc ttgggtggag gtggtgctgg ccctgcgcaa
     1501 catgaagaat ctgctgtttt ccgagatgtg ggatatcttc ctggtgcggc tgctgatggc
     1561 catggccgtg atgctgtact attctaactt tgtgctggcc ctggaggaga ggttcggcgt
     1621 gcgccctaag gtgacaggct acctgatctc ctatagctcc atgctgggag cagtggcagg
     1681 actggccctg ggaccaatcc tgcggctgta caagcacaat tctcaggccc tgctgctgca
     1741 ctctagcatc ctgacctgca cactgctgct gctgtatagc ctggcaccaa ccatgggagc
     1801 agtggtgctg tcctctaccc tgctgtcctt ttctacagcc atcggcagaa cctgtatcac
     1861 agacctgcag ctgaccgtgg gaggagcaca ggcaagcggc acactgatcg gagtgggaca
     1921 gtccgtgacc gcagtgggaa ggatcatcgc accactgctg agcggagtgg cacaggaggt
     1981 gtctccttgc ggaccaccta gcctgggagc cgtgctggcc ctggtggcca tcttcatcat
     2041 gtccctgaac aagagacaca gctccggcga tggcaattct aagctgaaga gcgaggaccc
     2101 agctttcttg tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga
     2161 acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag
     2221 tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct
     2281 ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta
     2341 cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg
     2401 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
     2461 gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct
     2521 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     2581 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
     2641 atggttactc accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc
     2701 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
     2761 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
     2821 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     2881 tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt
     2941 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     3001 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     3061 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     3121 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga
     3181 attggttaat tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag
     3241 ctcatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt
     3301 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3361 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3421 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     3481 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3541 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3601 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3661 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3721 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3781 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3841 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3901 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3961 tgctcacatg tt
//
