LOCUS       sequence-260094-        4008 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 4008)
  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 4008)
  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_SLC16A12
FEATURES             Location/Qualifiers
     source          1..4008
                     /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(2133..2232)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2249..2268)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2250..2268)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2273..2289)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2273..2289)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2391..2410)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2402..3211
                     /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(2469..2488)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3358..3946
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3847..3866
                     /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 accatggcca aggtgaacag ggcaagatct accagcccac ctgacggagg
      721 atggggatgg atgatcgtgg caggctgttt cctggtgacc atctgcacac gggccgtgac
      781 cagatgtatc agcatcttct ttgtggagtt ccagacctac tttacacagg actatgccca
      841 gaccgcctgg atccactcta tcgtggattg cgtgacaatg ctgtgcgcac cactgggaag
      901 cgtggtgtcc aatcacctga gctgtcaagt gggaatcatg ctgggaggac tgctggcaag
      961 caccggactg atcctgagct ccttcgccac atccctgaag cacctgtacc tgaccctggg
     1021 cgtgctgaca ggactgggat tcgccctgtg ctattctcca gccatcgcca tggtgggcaa
     1081 gtactttagc cggagaaagg ccctggccta tggcatcgcc atgtccggct ctggcatcgg
     1141 cacctttatc ctggcccccg tggtgcagct gctgatcgag cagttcagct ggaggggcgc
     1201 cctgctgatc ctgggaggct tcgtgctgaa cctgtgcgtg tgcggcgccc tgatgagacc
     1261 tatcaccctg aaggaggacc acaccacacc agagcagaat cacgtgtgca ggacacagaa
     1321 ggaggatatc aagcgcgtgt ccccttactc tagcctgacc aaggagtggg cccagacatg
     1381 cctgtgctgt tgcctgcagc aggagtatag cttcctgctg atgtccgact ttgtggtgct
     1441 ggccgtgtcc gtgctgttca tggcctacgg ctgctctcca ctgttcgtgt acctggtgcc
     1501 ctatgccctg tctgtgggcg tgagccacca gcaggcagcc ttcctgatgt ccatcctggg
     1561 cgtgatcgac atcatcggca acatcacctt tggctggctg acagataggc gctgtctgaa
     1621 gaattaccag tacgtgtgct acctgttcgc cgtgggcatg gacggcctgt gctatctgtg
     1681 cctgcctatg ctgcagtctc tgcctctgct ggtgcccttc agctgtacct tcggctactt
     1741 tgatggcgcc tatgtgacac tgatccccgt ggtgaccaca gagatcgtgg gcaccacaag
     1801 cctgtcctct gccctgggcg tggtgtactt cctgcacgcc gtgccttatc tggtgtcccc
     1861 accaatcgca ggcaggctgg tggataccac aggctcctac accgccgcct ttctgctgtg
     1921 cggcttctct atgatcttta gctccgtgct gctgggattc gcaaggctga tcaagaggat
     1981 gcgcaagacc cagctgcagt ttatcgccaa ggagtccgac ccaaagctgc agctgtggac
     2041 aaacggctcc gtggcctatt ctgtggcccg ggagctggat cagaagcacg gagagccagt
     2101 ggcaaccgca gtgcctggct actctctgac agacccagct ttcttgtaca aagttggcat
     2161 tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa
     2221 aatcattatt tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc
     2281 tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     2341 aataatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
     2401 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
     2461 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     2521 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     2581 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     2641 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     2701 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     2761 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     2821 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     2881 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     2941 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     3001 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     3061 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     3121 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     3181 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg
     3241 gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt
     3301 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3361 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3421 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3481 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     3541 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3601 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3661 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3721 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3781 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3841 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3901 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3961 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtt
//
