LOCUS       sequence-259093-        3915 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 3915)
  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 3915)
  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_MFSD10
FEATURES             Location/Qualifiers
     source          1..3915
                     /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(2040..2139)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2156..2175)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2157..2175)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2180..2196)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2180..2196)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2298..2317)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2309..3118
                     /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(2376..2395)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3265..3853
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3754..3773
                     /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 accatgggat ggggaggagg aggaggatgt accccaaggc cacctatcca
      721 ccagcagcca ccagagcgga gagtggtgac cgtggtgttt ctgggcctgc tgctggacct
      781 gctggccttc acactgctgc tgccactgct gcctggactg ctggagtctc acggaagggc
      841 acacgaccct ctgtacggaa gctggcaggg aggcgtggat tggtttgcca cagccatcgg
      901 catgccagtg gagaagcggt acaacagcgt gctgttcgga ggactgatcg gatccgcctt
      961 ttctgtgctg cagttcctgt gcgcaccact gaccggagcc acaagcgatt gtctgggcag
     1021 gcgccctgtg atgctgctgt gcctgatggg agtggccacc tcctacgccg tgtgggccac
     1081 aagccggtcc tttgccgcct tcctggcctc tagactgatc ggcggcatca gcaagggcaa
     1141 cgtgagcctg agcaccgcaa tcgtggcaga cctgggaagc ccactggcac ggtcccaggg
     1201 catggccgtg atcggcgtgg ccttttccct gggcttcaca ctgggcccta tgctgggagc
     1261 atctctgcct ctggagatgg caccatggtt tgccctgctg ttcgccgcct ctgatctgct
     1321 gttcatcttt tgcttcctgc cagagaccct gcccctggag aagagggcac catccatcgc
     1381 cctgggcttt cgcgacgcag cagatctgct gagcccactg gccctgctgc ggttctccgc
     1441 cgtggcaaga ggacaggacc ctccaagcgg cgataggctg agctccctgc ggagactggg
     1501 actggtgtac tttctgtatc tgtttctgtt ctccggcctg gagtacaccc tgtctttcct
     1561 gacacaccag agatttcagt tctctagcct gcagcagggc aagatgttct ttctgatcgg
     1621 cctgaccatg gccacaatcc agggagcata tgcaaggagg atccacccag gaggagaggt
     1681 ggcagcagtg aagcgggccc tgctgctgct ggtgccagcc tttctgctga tcggctgggg
     1741 cagaagcctg cccgtgctgg gactgggact gctgctgtac tccttcgccg cagcagtggt
     1801 ggtgccttgt ctgtcctctg tggtggccgg ctatggctct ccaggccaga agggcaccgt
     1861 gatgggcaca ctgaggagcc tgggcgccct ggcaagggca gcaggacctc tggtggcagc
     1921 atccgtgtac tggctggcag gagcacaggc atgctttacc acatggtccg gcctgttcct
     1981 gctgcccttc tttctgctgc agaagctgtc ttatcctgcc cagaccctga aggccgagga
     2041 cccagctttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa
     2101 cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg atatccccta
     2161 tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa
     2221 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
     2281 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc
     2341 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     2401 cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt
     2461 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     2521 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     2581 tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata
     2641 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     2701 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
     2761 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     2821 gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt
     2881 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     2941 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     3001 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     3061 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     3121 agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc
     3181 aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc
     3241 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3301 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3361 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     3421 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3481 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3541 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3601 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3661 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3721 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3781 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3841 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3901 ttttgctcac atgtt
//
