LOCUS       sequence-258982-        4020 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 4020)
  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 4020)
  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_MFSD14A
FEATURES             Location/Qualifiers
     source          1..4020
                     /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(2145..2244)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2261..2280)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2262..2280)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2285..2301)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2285..2301)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2403..2422)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2414..3223
                     /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(2481..2500)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3370..3958
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3859..3878
                     /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 accatgaccc agggcaagaa gaagaagcgg gccgccaaca gatccatcat
      721 gctggccaag aagatcatca tcaaggacgg aggaacacca cagggaatcg gatcccctag
      781 cgtgtaccac gccgtgatcg tgatctttct ggagttcttt gcatggggac tgctgaccgc
      841 acctacactg gtggtgctgc acgagacctt tccaaagcac acattcctga tgaatggcct
      901 gatccagggc gtgaagggcc tgctgtcctt tctgtctgcc ccactgatcg gcgccctgtc
      961 cgacgtgtgg ggaaggaagt ctttcctgct gctgaccgtg ttctttacat gcgccccaat
     1021 ccccctgatg aagatctctc cctggtggta tttcgccgtg atcagcgtgt ccggcgtgtt
     1081 cgccgtgacc tttagcgtgg tgttcgccta cgtggccgat atcacacagg agcacgagag
     1141 gagcatggcc tatggcctgg tgtccgccac ctttgccgcc tctctggtga caagccctgc
     1201 aatcggagca tacctgggac gcgtgtatgg cgactccctg gtggtggtgc tggcaaccgc
     1261 aatcgccctg ctggatatct gcttcatcct ggtggccgtg cctgagtctc tgccagagaa
     1321 gatgaggcca gcatcttggg gagcaccaat cagctgggag caggcagacc cctttgcctc
     1381 tctgaagaag gtcggccagg atagcatcgt gctgctgatc tgtatcacag tgttcctgtc
     1441 ctacctgccc gaggccggcc agtacagctc cttctttctg tatctgcggc agatcatgaa
     1501 gtttagccct gagtccgtgg cagccttcat cgccgtgctg ggcatcctga gcatcatcgc
     1561 ccagaccatc gtgctgagcc tgctgatgag atccatcggc aacaagaata caatcctgct
     1621 gggcctgggc tttcagatcc tgcagctggc ctggtatggc ttcggcagcg agccatggat
     1681 gatgtgggca gcaggagcag tggcagcaat gtctagcatc acctttccag ccgtgtccgc
     1741 cctggtgtct aggacagcag acgcagatca gcagggagtg gtgcagggca tgatcaccgg
     1801 catcagagga ctgtgcaacg gactgggacc tgccctgtac ggcttcatct tttatatctt
     1861 tcacgtggag ctgaaggagc tgccaatcac cggcacagac ctgggcacca acacatcccc
     1921 ccagcaccac ttcgagcaga attctatcat ccctggcccc cctttcctgt ttggagcatg
     1981 cagcgtgctg ctggccctgc tggtggccct gttcatcccc gagcacacca atctgtccct
     2041 gaggtcctct agctggcgca agcactgtgg ctctcacagc caccctcaca acacccaggc
     2101 accaggagag gcaaaggagc cactgctgca ggatacaaat gtggacccag ctttcttgta
     2161 caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat
     2221 cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta
     2281 catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat
     2341 tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa
     2401 tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa
     2461 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     2521 gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     2581 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     2641 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     2701 cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     2761 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     2821 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     2881 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     2941 ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     3001 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     3061 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     3121 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     3181 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg
     3241 gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa
     3301 atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3361 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     3421 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     3481 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     3541 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     3601 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     3661 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     3721 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     3781 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     3841 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     3901 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     3961 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
//
