LOCUS       sequence-259553-        3963 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 3963)
  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 3963)
  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_SLC16A11
FEATURES             Location/Qualifiers
     source          1..3963
                     /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"
     regulatory      1529..1538
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2088..2187)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2204..2223)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2205..2223)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2228..2244)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2228..2244)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2346..2365)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2357..3166
                     /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(2424..2443)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3313..3901
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3802..3821
                     /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 accatgcctg ccccacagag aaagcaccgg agaggcggct tctcccacag
      721 gtgctttccc acccctcaga cagccatgac cccacagcca gcaggaccac ctgacggagg
      781 atggggctgg gtggtggccg ccgccgcctt cgccatcaac ggcctgtcct acggactgct
      841 gcgctctctg ggactggcat tcccagatct ggcagagcac tttgaccggt ctgcccagga
      901 tacagcctgg atcagcgccc tggccctggc cgtgcagcag gccgccagcc ccgtgggcag
      961 cgccctgtcc accagatggg gagcaaggcc tgtggtcatg gtgggaggcg tgctggcatc
     1021 cctgggcttc gtgttttccg ccttcgcctc tgacctgctg cacctgtacc tgggactggg
     1081 actgctggca ggattcggat gggcactggt gtttgcacct gccctgggca cactgagccg
     1141 ctatttctcc aggcgccggg tgctggcagt gggactggcc ctgaccggca atggagcaag
     1201 ctccctgctg ctggcacccg ccctgcagct gctgctggac acatttggct ggagaggcgc
     1261 cctgctgctg ctgggagcca tcacactgca cctgacccca tgcggcgccc tgctgctgcc
     1321 tctggtgctg ccaggcgatc caccagcacc tccaaggtcc ccactggccg ccctgggcct
     1381 gtctctgttc acaagaaggg ccttcagcat ctttgcactg ggaaccgccc tggtgggagg
     1441 aggatacttt gtgccctatg tgcacctggc ccctcacgcc ctggataggg gactgggagg
     1501 atacggagcc gccctggtgg tggccgtggc cgccatgggc gacgccggcg ccagactggt
     1561 gtgcggatgg ctggcagatc agggatgggt gcctctgcca aggctgctgg ccgtgttcgg
     1621 cgccctgaca ggactgggac tgtgggtggt gggactggtg cctgtggtgg gaggcgagga
     1681 gtcttgggga ggaccactgc tggcagcagc agtggcctac ggcctgtctg ccggcagcta
     1741 tgcaccactg gtgtttggcg tgctgccagg actggtggga gtgggaggcg tggtgcaggc
     1801 caccggcctg gtcatgatgc tgatgtccct gggaggactg ctgggaccac ctctgtctgg
     1861 cttcctgcgc gacgagacag gcgatttcac cgccagcttt ctgctgtctg gcagcctgat
     1921 cctgagcggc tcttttatct atatcggact gccacgggcc ctgccttctt gcggcccagc
     1981 cagcccaccc gccacacctc cacccgagac cggagagctg ctgccagcac ctcaggccgt
     2041 gctgctgagc ccaggcggcc ccggctccac cctggacacc acatgtgacc cagctttctt
     2101 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     2161 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta
     2221 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     2281 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     2341 taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc
     2401 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
     2461 tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg
     2521 caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga
     2581 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     2641 caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     2701 tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     2761 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     2821 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     2881 agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg
     2941 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     3001 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     3061 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     3121 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa
     3181 ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc
     3241 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     3301 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3361 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3421 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3481 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3541 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3601 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3661 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3721 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3781 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3841 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3901 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     3961 gtt
//
