LOCUS       sequence-260453-        4050 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 4050)
  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 4050)
  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_SLC24A5
FEATURES             Location/Qualifiers
     source          1..4050
                     /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(2175..2274)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2291..2310)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2292..2310)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2315..2331)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2315..2331)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2433..2452)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2444..3253
                     /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(2511..2530)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3400..3988
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3889..3908
                     /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 accatgcaga ccaagggagg acagacatgg gcaaggagag ccctgctgct
      721 gggaatcctg tgggcaaccg cacacctgcc actgtctggc acaagcctgc cacagaggct
      781 gccaagggca accggaaact ctacacagtg cgtgatcagc cctagctccg agttcccaga
      841 gggcttcttt accaggcagg agaggaggga cggaggaatc atcatctact tcctgatcat
      901 cgtgtatatg tttatggcca tctccatcgt gtgcgacgag tacttcctgc catctctgga
      961 gatcatctcc gagtctctgg gactgagcca ggatgtggca ggaaccacct tcatggcagc
     1021 aggatctagc gccccagagc tggtgaccgc cttcctgggc gtgtttatca caaagggcga
     1081 tatcggcatc tctaccatcc tgggcagcgc catctataac ctgctgggaa tctgcgcagc
     1141 atgtggactg ctgagcaata ccgtgtccac actgtcttgc tggcctctgt tcagggactg
     1201 tgcagcatac accatctccg ccgcagccgt gctgggcatc atctatgata accaggtgta
     1261 ctggtatgag ggcgccctgc tgctgctgat ctacggcctg tatgtgctgg tgctgtgctt
     1321 tgacatcaag atcaatcagt acatcatcaa gaagtgcagc ccatgctgtg cctgtctggc
     1381 caaggccatg gagaggtccg agcagcagcc actgatggga tgggaggatg agggccagcc
     1441 tttcatccgg agacagtccc gcaccgactc tggcatcttt tacgaggata gcggctattc
     1501 ccagctgagc atctccctgc acggactgtc tcaggtgagc gaggacccac ctagcgtgtt
     1561 caacatgcct gaggccgatc tgaagcggat cttttgggtg ctgtccctgc caatcatcac
     1621 actgctgttc ctgaccacac ccgactgcag aaagaagttc tggaagaact acttcgtgat
     1681 cacattcttt atgtccgcca tctggatctc tgcctttacc tatatcctgg tgtggatggt
     1741 gaccatcaca ggcgagaccc tggagatccc cgacaccgtg atgggactga cactgctggc
     1801 agcaggcacc tccatccctg atacaatcgc ctctgtgctg gtggccagga agggcaaggg
     1861 cgacatggcc atgagcaaca tcgtgggctc caacgtgttc gatatgctgt gcctgggcat
     1921 cccttggttc atcaagacag cctttatcaa cggcagcgcc ccagcagaag tgaattccag
     1981 gggactgacc tacatcacaa tcagcctgaa catctccatc atcttcctgt ttctggccgt
     2041 gcactttaat ggctggaagc tggatagaaa gctgggcatc gtgtgcctgc tgtcttacct
     2101 gggcctggcc accctgagcg tgctgtatga gctgggcatc atcggcaaca ataagatccg
     2161 cggctgtggc ggcgacccag ctttcttgta caaagttggc attataagaa agcattgctt
     2221 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     2281 agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
     2341 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac
     2401 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
     2461 ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     2521 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga
     2581 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     2641 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     2701 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca
     2761 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     2821 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     2881 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     2941 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
     3001 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     3061 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     3121 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     3181 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     3241 tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac
     3301 ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     3361 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3421 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3481 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3541 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3601 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3661 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3721 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3781 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3841 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3901 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     3961 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4021 ggccttttgc tggccttttg ctcacatgtt
//
