LOCUS       sequence-260409-        4416 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 4416)
  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 4416)
  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_SLC24A4
FEATURES             Location/Qualifiers
     source          1..4416
                     /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(2541..2640)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2657..2676)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2658..2676)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2681..2697)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2681..2697)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2799..2818)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2810..3619
                     /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(2877..2896)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3766..4354
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4255..4274
                     /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 accatggccc tgagaggcac cctgaggcca ctgaaggtgc ggagaaggcg
      721 cgagatgctg ccacagcaag tgggattcgt gtgcgccgtg ctggccctgg tgtgctgtgc
      781 ctctggcctg tttggaagcc tgggacacaa gaccgcatcc gcctctaagc gggtgctgcc
      841 tgacacatgg cggaacagaa agctgatggc cccagtgaac ggcacccaga cagccaagaa
      901 ttgtaccgac cctgccatcc acgagttccc aacagatctg tttagcaata aggagagaca
      961 gcacggagcc gtgctgctgc acatcctggg cgccctgtac atgttctatg ccctggccat
     1021 cgtgtgcgac gatttctttg tgccctctct ggagaagatc tgtgagaggc tgcacctgag
     1081 cgaggacgtg gcaggagcca cctttatggc agcaggcagc tccacacctg agctgttcgc
     1141 cagcgtgatc ggcgtgttta tcacccacgg cgatgtggga gtgggaacaa tcgtgggatc
     1201 cgccgtgttc aacatcctgt gcatcatcgg cgtgtgcgga ctgtttgcag gacaggtggt
     1261 gcgcctgacc tggtgggccg tgtgccggga cagcgtgtac tatacaatca gcgtgatcgt
     1321 gctgatcgtg ttcatctacg atgagcagat cgtgtggtgg gagggcctgg tgctgatcat
     1381 cctgtacgtg ttttatatcc tgatcatgaa gtataacgtg aagatgcagg ccttctttac
     1441 cgtgaagcag aagagcatcg ccaacggcaa tcctgtgaac tccgagctgg aggccggcaa
     1501 tgacttctac gatggctcct atgacgatcc atctgtgccc ctgctgggcc aggtgaagga
     1561 gaagccccag tacggcaaga atcctgtggt catggtggac gagatcatgt ctagctcccc
     1621 ccctaagttc acatttcccg aggccggcct gagaatcatg atcaccaaca agttcggacc
     1681 taggacaagg ctgaggatgg catccaggat catcatcaat gagagacaga ggctgatcaa
     1741 ctctgccaat ggcgtgtcta gcaagccact gcagaacggc agacacgaga atatcgagaa
     1801 cggcaatgtg cccgtggaga accctgagga tccacagcag aatcaggagc agcagccacc
     1861 acctcagcca ccacctccag agccagagcc tgtggaggca gacttcctga gccccttttc
     1921 cgtgcctgag gcaaggggcg ataaggtgaa gtgggtgttc acctggcccc tgatctttct
     1981 gctgtgcgtg acaatcccaa actgttctaa gccccggtgg gagaagttct ttatggtgac
     2041 cttcatcacc gccacactgt ggatcgccgt gttttcctac atcatggtgt ggctggtgac
     2101 catcatcggc tatacactgg gcatccctga cgtgatcatg ggcatcacct tcctggcagc
     2161 aggcacatcc gtgccagatt gtatggcctc tctgatcgtg gcaaggcagg gactgggcga
     2221 catggccgtg agcaacacca tcggctccaa cgtgttcgat atcctggtgg gactgggagt
     2281 gccatgggga ctgcagacca tggtggtgaa ctacggctcc acagtgaaga tcaattctcg
     2341 gggcctggtg tatagcgtgg tgctgctgct gggaagcgtg gccctgaccg tgctgggaat
     2401 ccacctgaat aagtggcgcc tggatcggaa gctgggcgtg tacgtgctgg tgctgtatgc
     2461 catcttcctg tgcttttcca tcatgatcga gttcaacgtg ttcacatttg tgaatctgcc
     2521 catgtgcagg gaggacgatg acccagcttt cttgtacaaa gttggcatta taagaaagca
     2581 ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg
     2641 ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca
     2701 gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     2761 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     2821 tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     2881 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa
     2941 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     3001 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     3061 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc
     3121 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     3181 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     3241 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     3301 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt
     3361 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     3421 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     3481 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     3541 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     3601 gctcgatgag tttttctaat cagaattggt taattggttg taacactggc agagcattac
     3661 gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc
     3721 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3781 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3841 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3901 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3961 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4021 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4081 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4141 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4201 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4261 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4321 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4381 gttcctggcc ttttgctggc cttttgctca catgtt
//
