LOCUS       sequence-259277-        4011 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 4011)
  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 4011)
  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_SLC16A4
FEATURES             Location/Qualifiers
     source          1..4011
                     /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(2136..2235)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2252..2271)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2253..2271)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2276..2292)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2276..2292)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2394..2413)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2405..3214
                     /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(2472..2491)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3361..3949
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3850..3869
                     /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 accatgctga agcgggaggg caaggtgcag ccatacacca agacactgga
      721 cggcggctgg ggctggatga tcgtgatcca cttctttctg gtgaacgtgt ttgtgatggg
      781 catgaccaag acattcgcca tcttctttgt ggtgtttcag gaggagttcg agggcaccag
      841 cgagcagatc ggctggatcg gctccatcat gagctccctg agattctgcg caggaccact
      901 ggtggcaatc atctgtgata tcctgggcga gaagaccaca agcatcctgg gcgcctttgt
      961 ggtgacaggc ggctacctga tctctagctg ggccaccagc atccccttcc tgtgcgtgac
     1021 aatgggactg ctgcctggac tgggatccgc ctttctgtac caggtggccg ccgtggtgac
     1081 cacaaagtat ttcaagaagc ggctggccct gtccaccgca atcgcaagat ctggcatggg
     1141 cctgaccttt ctgctggccc cctttacaaa gttcctgatc gacctgtatg attggacagg
     1201 cgccctgatc ctgttcggag caatcgccct gaatctggtg ccttcctcta tgctgctgag
     1261 gccaatccac atcaagagcg agaacaattc cggcatcaag gacaagggaa gctccctgtc
     1321 tgcccacgga cctgaggcac acgcaaccga gacacactgc cacgagaccg aggagtctac
     1381 aatcaaggat agcaccacac agaaggccgg cctgccatcc aagaacctga ccgtgtctca
     1441 gaatcagagc gaggagttct acaacggccc caaccggaat agactgctgc tgaagtccga
     1501 cgaggagtct gataaagtga tctcctggtc ttgtaagcag ctgtttgaca tcagcctgtt
     1561 caggaatcct ttcttttaca tctttacctg gagcttcctg ctgtcccagc tggcctactt
     1621 catcccaacc ttccacctgg tggcaagggc aaagacactg ggcatcgaca tcatggatgc
     1681 ctcttacctg gtgagcgtgg caggaatcct ggagaccgtg tcccagatca tctctggctg
     1741 ggtggccgat cagaactgga tcaagaagta ccactatcac aagagctatc tgatcctgtg
     1801 cggcatcaca aatctgctgg ccccactggc caccacattt cccctgctga tgacctacac
     1861 aatctgcttt gccatcttcg caggaggata tctggccctg atcctgcctg tgctggtgga
     1921 cctgtgcagg aactccaccg tgaatcgctt cctgggcctg gcctctttct ttgcaggaat
     1981 ggccgtgctg agcggaccac ctatcgccgg ctggctgtac gattataccc agacatacaa
     2041 cggcagcttt tatttctccg gcatctgtta tctgctgtct agcgtgtcct tctttttcgt
     2101 gcccctggcc gagcggtgga agaattctct gaccgaccca gctttcttgt acaaagttgg
     2161 cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa
     2221 taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat
     2281 agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga
     2341 taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg
     2401 tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc
     2461 tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta
     2521 tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt
     2581 tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct
     2641 tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat
     2701 ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt
     2761 tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt
     2821 taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt
     2881 tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga
     2941 aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact
     3001 tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg
     3061 aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc
     3121 ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt
     3181 gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca
     3241 ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa
     3301 cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3361 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     3421 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3481 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     3541 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3601 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3661 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3721 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     3781 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     3841 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3901 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3961 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt t
//
