LOCUS       sequence-263863-        3837 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 3837)
  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 3837)
  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_SLC30A4
FEATURES             Location/Qualifiers
     source          1..3837
                     /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(1962..2061)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2078..2097)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2079..2097)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2102..2118)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2102..2118)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2220..2239)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2231..3040
                     /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(2298..2317)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3187..3775
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3676..3695
                     /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 accatggcag gaagcggagc atggaagagg ctgaagtcca tgctgcgcaa
      721 ggacgatgcc cctctgtttc tgaacgatac cagcgccttc gacttcagcg acgaggcagg
      781 cgacgaggga ctgtcccggt tcaacaagct gagagtggtg gtggcagacg atggatctga
      841 ggcaccagag aggcctgtga atggagcaca ccccaccctg caggcagacg atgactccct
      901 gctggatcag gacctgcctc tgacaaactc ccagctgtct ctgaaggtgg attcctgcga
      961 caattgttct aagcagagag agatcctgaa gcagaggaag gtgaaggcaa ggctgaccat
     1021 cgcagccgtg ctgtacctgc tgtttatgat cggcgagctg gtgggcggct atatcgccaa
     1081 tagcctggcc atcatgaccg atgccctgca catgctgaca gacctgtccg ccatcatcct
     1141 gacactgctg gccctgtggc tgagctccaa gtctccaacc aagaggttca catttggctt
     1201 ccacaggctg gaggtgctga gcgccatgat ctccgtgctg ctggtgtaca tcctgatggg
     1261 ctttctgctg tatgaggccg tgcagaggac catccacatg aactacgaga tcaatggcga
     1321 tatcatgctg atcacagcag cagtgggagt ggcagtgaac gtgatcatgg gcttcctgct
     1381 gaatcagagc ggacacaggc actctcacag ccactccctg ccatctaaca gcccaaccag
     1441 gggatctgga tgcgagagaa atcacggaca ggatagcctg gccgtgcggg cagcctttgt
     1501 gcacgcactg ggcgacctgg tgcagtccgt gggcgtgctg atcgcagcct acatcatccg
     1561 gttcaagcct gagtataaga tcgccgaccc aatctgtacc tacgtgttca gcctgctggt
     1621 ggcctttacc acattcagga tcatctggga tacagtggtc atcatcctgg agggcgtgcc
     1681 aagccacctg aacgtggact acatcaagga ggccctgatg aagatcgagg acgtgtactc
     1741 cgtggaggac ctgaatatct ggagcctgac ctccggcaag tctacagcca tcgtgcacat
     1801 ccagctgatc cccggctcta gctccaagtg ggaggaggtg cagtctaagg ccaaccacct
     1861 gctgctgaat accttcggca tgtacagatg cacaatccag ctgcagagct atcggcagga
     1921 ggtggataga acatgcgcca actgtcagtc tagctccccc gacccagctt tcttgtacaa
     1981 agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag
     2041 tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat
     2101 ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc
     2161 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
     2221 aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat
     2281 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     2341 aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     2401 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     2461 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     2521 tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     2581 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     2641 tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
     2701 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     2761 gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt
     2821 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     2881 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     2941 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     3001 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     3061 gtaacactgg cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc
     3121 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3181 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3241 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3301 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3361 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3421 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3481 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3541 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3601 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3661 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3721 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3781 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
