LOCUS       sequence-259255-        4203 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 4203)
  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 4203)
  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_SLC22A4
FEATURES             Location/Qualifiers
     source          1..4203
                     /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      1196..1205
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2328..2427)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2444..2463)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2445..2463)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2468..2484)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2468..2484)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2586..2605)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2597..3406
                     /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(2664..2683)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3553..4141
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4042..4061
                     /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 accatgcggg actacgatga agtgatcgcc tttctgggcg agtggggccc
      721 tttccagaga ctgatcttct ttctgctgtc tgccagcatc atcccaaacg gctttaatgg
      781 catgtctgta gtatttcttg caggaacccc tgagcacagg tgcagagtgc cagacgcagc
      841 aaacctgagc tccgcctgga ggaacaatag cgtgccactg aggctgcgcg atggcagaga
      901 ggtgccccac tcctgttctc ggtatagact ggccacaatc gccaattttt ccgccctggg
      961 actggagcca ggaagggacg tggatctggg acagctggag caggagtctt gcctggacgg
     1021 ctgggagttc tctcaggacg tgtacctgag caccgtggtg acagagtgga acctggtgtg
     1081 cgaggacaat tggaaggtgc ccctgaccac atctctgttc tttgtgggcg tgctgctggg
     1141 ctcttttgtg agcggccagc tgagcgatcg gtttggcaga aagaacgtgc tgttcgccac
     1201 catggccgtg cagacaggct tcagctttct gcagatcttc agcatctcct gggagatgtt
     1261 caccgtgctg tttgtgatcg tgggcatggg ccagatctcc aattatgtgg tggcctttat
     1321 cctgggcaca gagatcctgg gcaagtccgt gaggatcatc ttctctaccc tgggcgtgtg
     1381 cacattcttt gccgtgggct acatgctgct gcctctgttc gcctacttca tcagggactg
     1441 gaggatgctg ctgctggccc tgaccgtgcc aggcgtgctg tgcgtgccac tgtggtggtt
     1501 tatccccgag agccctaggt ggctgatctc ccagcggaga ttccgcgagg ccgaggatat
     1561 catccagaag gccgccaaga tgaacaatat cgccgtgcct gccgtgatct tcgactccgt
     1621 ggaggagctg aacccactga agcagcagaa ggccttcatc ctggatctgt ttaggacccg
     1681 caatatcgcc atcatgacaa tcatgtctct gctgctgtgg atgctgacct ctgtgggcta
     1741 ctttgccctg agcctggacg ccccaaacct gcacggcgat gcctacctga attgcttcct
     1801 gagcgccctg atcgagatcc ccgcctatat caccgcctgg ctgctgctga ggacactgcc
     1861 taggcgctac atcatcgcag ccgtgctgtt ttggggagga ggcgtgctgc tgttcatcca
     1921 gctggtgccc gtggactact attttctgag catcggcctg gtcatgctgg gcaagttcgg
     1981 catcacatcc gccttttcta tgctgtacgt gttcaccgcc gagctgtatc ctacactggt
     2041 gaggaacatg gcagtgggag tgaccagcac agcatccagg gtgggatcca tcatcgcacc
     2101 atacttcgtg tatctgggcg cctacaatcg catgctgccc tatatcgtga tgggcagcct
     2161 gaccgtgctg atcggcatcc tgacactgtt ctttcccgag tccctgggca tgaccctgcc
     2221 tgagacactg gagcagatgc agaaggtgaa gtggtttcgg tccggcaaga agaccagaga
     2281 ttctatggag acagaggaga accccaaggt gctgatcacc gccttcgacc cagctttctt
     2341 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     2401 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta
     2461 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     2521 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     2581 taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc
     2641 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
     2701 tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg
     2761 caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga
     2821 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     2881 caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     2941 tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     3001 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     3061 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     3121 agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg
     3181 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     3241 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     3301 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     3361 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa
     3421 ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc
     3481 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     3541 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3601 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3661 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3721 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3781 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3841 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3901 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3961 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4021 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4081 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4141 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4201 gtt
//
