LOCUS       sequence-259258-        3786 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 3786)
  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 3786)
  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_MFSD3
FEATURES             Location/Qualifiers
     source          1..3786
                     /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(1911..2010)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2027..2046)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2028..2046)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2051..2067)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2051..2067)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2169..2188)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2180..2989
                     /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(2247..2266)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3136..3724
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3625..3644
                     /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 accatgaggg gcaagctgct gcctctggca ggactgtacc tggtgcaggg
      721 actgccatat ggactgcagt ccggactgct gcctgtgctg ctgagggcag gcggcctgtc
      781 tctgaccaga gtgggcctgg ccaaggtgct gtacgcacca tggctgctga agctggcatg
      841 ggcaccactg gtggacgcac agggcagcgc cagggcatgg gtgaccagat ccacagcagg
      901 actgggactg gtgtgcggcc tgctggccgg cctgccccct ccaggagcag gacaggcagg
      961 actgccagca gcagtggcag gactgctgct gctgctgaac ctgggagcag caatgcagga
     1021 cgtggccctg gatgcactgg cagtgcagct gctggagcct gcagagctgg gcccaggcaa
     1081 taccgtgcag gtggtggcct acaagctggg agccgccctg gcaggaggcg ccctgctggc
     1141 cctgctgccc accttcagct ggcctcagct gtttctgctg ctggcagcaa catattggct
     1201 ggcagccgcc ctggcatggg cagcaccagc cctgcggaga ctgccacagc agccaccttc
     1261 cgagcagagg ccacacaccg cacacctgct gagggacgtg ctggcagtgc ctggaaccgt
     1321 gtggacagca ggcttcgtgc tgacatataa gctgggagag cagggagcaa gctccctgtt
     1381 tcctctgctg ctgctggatc acggcgtgtc tgccccagag ctgggactgt ggaacggagt
     1441 gggagcagtg gtgtgcagca tcgcaggatc tagcctggga ggcaccctgc tggcaaagca
     1501 ctggaagctg ctgccactgc tgcggtccgt gctgaggttc cgcctgggag gactggcatg
     1561 tcagaccgcc ctggtgtttc acctggacac actgggcgcc tctatggatg ccggcacaat
     1621 cctgagaggc tccgccctgc tgtctctgtg cctgcagcac ttcctgggcg gcctggtgac
     1681 cacagtgacc tttacaggca tgatgaggtg tagccagctg gcaccacgcg ccctgcaggc
     1741 aacccactac tccctgctgg ccacactgga gctgctgggc aagctgctgc tgggcaccct
     1801 ggcaggagga ctggcagacg gactgggacc acacccctgt ttcctgctgc tgctgatcct
     1861 gtctgccttt cctgtgctgt atctggatct ggccccaagc acattcctgg acccagcttt
     1921 cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt
     1981 cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc
     2041 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     2101 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     2161 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     2221 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     2281 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     2341 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     2401 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     2461 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     2521 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     2581 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     2641 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     2701 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     2761 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     2821 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     2881 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     2941 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     3001 taattggttg taacactggc agagcattac gctgacttga cgggacggcg caagctcatg
     3061 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     3121 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3181 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3241 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3301 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3361 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3421 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3481 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3541 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3601 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3661 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3721 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     3781 catgtt
//
