LOCUS       sequence-333104-        4221 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors. Contains a STOP codon at the end of 
            the codon-optimized ORF sequence. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4221)
  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 4221)
  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-SLC22A3_STOP
FEATURES             Location/Qualifiers
     source          1..4221
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(25..42)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(196..784)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(276..295)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(931..1740)
                     /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     1654..1673
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1732..1751
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1853..1869
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1853..1869
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1874..1893
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1874..1892
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1910..2009
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3687..3786)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3802..3819)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3802..3818)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3811..3833)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      3882..3968
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4060..4087
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
       61 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
      121 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      181 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      241 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      301 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      361 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
      421 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
      481 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
      541 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
      601 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
      661 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
      721 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
      781 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg acgcgtaact
      841 cacgttaagg gattttggtc atgagcttgc gccgtcccgt caagtcagcg taatgctctg
      901 ccagtgttac aaccaattaa ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa
      961 ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa
     1021 tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc
     1081 gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt
     1141 atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagtttatg
     1201 catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc
     1261 atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct
     1321 gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc
     1381 atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg ctgtttttcc
     1441 ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt
     1501 cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt
     1561 ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa
     1621 gcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa
     1681 atcagcatcc atgttggaat ttaatcgcgg cctcgacgtt tcccgttgaa tatggctcat
     1741 aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt
     1801 tttatcttgt gcaatgtaac atcagagatt ttgagacacg ggccagagct gccaggaaac
     1861 agctatgacc atgtaatacg actcactata ggggatatca gctggatggc aaataatgat
     1921 tttattttga ctgatagtga cctgttcgtt gcaacaaatt gataagcaat gctttcttat
     1981 aatgccaact ttgtacaaga aagctgggtc ttacaggtga gagcgggaca cgggggtctt
     2041 cttattcctg ccacacttgc aggagtgtgg gctgcccagc ttctccacat cgtccactgt
     2101 ctctggcagg gcgatgccct tggtctcagg cagcagcatg accaggccgc cacagatgga
     2161 ggccaggatg ccgaagatga tcagtggcag ctccagccac acggctgcca ggcggaacag
     2221 cagaaaaggg gcgatgatgc cgccgaaatc gcacaggcca gagcacaggg acacgccaaa
     2281 attcctcagt gtggtgggat acagctcgga gttcaccagg tacacgatct caaaggccat
     2341 tgtgatgccc agtctgccca gggtggccac tgtggtcctc agccatgcga ttccctctgg
     2401 caggaaggct gtcaccaggc atgccactcc tgccacgata ttgcttgcgg caaaaggcag
     2461 tctccggccc agtctctcga tggtcagcag gatcagcagg gcgcctggca gctccaccac
     2521 tccgctgata aagaaatcga tatacaggtt gccgccgatg atgcccagcc gcatgaccag
     2581 gccctggtac accacggcag aggtgaacca ggcaaacatc aggatcagtg tacacttgcg
     2641 catctgaggg gtcctcacca gatccaggaa gctggggtta gacacctcct cgtctgtcac
     2701 ggtgatctcg ctataattgg agctcaggta cttgccgttg cacttggcga tgcgcctcag
     2761 gatctgcagg gccttgtcgc ccttctttct tgtgatcagc cacctagggg actctggcac
     2821 cacccaatag tacagcagga acagaaagct gggcagggtg atggccagct ggatgccctg
     2881 ccaattaggg atgaagtagg cgatgccggg caggatgatg atgcccagtg taaagaacat
     2941 ctggatcacg atgcccacga ttctccgctg cttggagccc acgatctcgg tcacgatcac
     3001 gtaacatgtc atccaggtgc ccttgccgaa cacgccctgc aggaaccgaa agatcacgaa
     3061 cactggaaag ttaggggcaa aggccaccac cactccggtc actcccactc ccaggcaaga
     3121 cagcaggtag atcacgattc tgccataccg gtctgctgcg tatcccagtg tgaaggctcc
     3181 ggtcagaaag cccagattca ggatggcctg tgtcagatcc agcatccagg cgttcacgca
     3241 caccaggtcg aactcgctca cgatggtaga gtgtgcctgt gcatatctcc atcctcccct
     3301 acatggcacc agtggggcag acctatttgg aaaggctgcc agaggatctg cgcaggacag
     3361 ggcgctggtg gcgctggcag agtcgtttgc tgcctccagc aggtaccgct ggcacctgcc
     3421 cctcctctca ggtggctctg gtcccctgga tgctggtgct gtcctgttcc actcctcctc
     3481 gggagaccag ccacatctct ctgccagggc ggcggcggat ggtcccctgc accagtagtg
     3541 atctggctgg gttcccagga acaccactcc cacgaacaga aaggcgaatg tcacgccggt
     3601 caggcacagc agcaggaaca ctctccgctg gaatctgcca aactctccca ccctctgcag
     3661 ggcctcgtca aagctaggca tggtggagcc tgcttttttg tacaaagttg gcattataaa
     3721 aaagcattgc tcatcaatgt gttgcaacga acaggtcact atcagtcaaa ataaaatcat
     3781 tatttggggc ccgagcttaa gactggccgt cgttttacaa cgtcgtgact gggaaaacat
     3841 ccatgctagc gttaacgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc
     3901 agtcggaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta
     3961 ggacaaatcc gccgggagcg gatttgaacg ttgtgaagca acggcccgga gggtggcggg
     4021 caggacgccc gccataaact gccaggcatc aaactaagca gaaggccatc ctgacggatg
     4081 gcctttttgc gtttctacaa actcttcctg gctagcggta cgcgtattaa ttgcgttgcg
     4141 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4201 acgcgcgggg agaggcggtt t
//
