LOCUS       sequence-260215-        4482 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 4482)
  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 4482)
  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_SLC24A3
FEATURES             Location/Qualifiers
     source          1..4482
                     /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"
     CDS             complement(1397..1414)
                     /codon_start=1
                     /product="HQHQHQ metal affinity tag"
                     /label=HQ tag
                     /note="The HQ tag binds to nickel and other metal ions much
                     like the 6xHis tag, but may allow elution at lower 
                     concentrations of imidazole (Godat et al., 2008)."
                     /translation="HQHQHQ"
     protein_bind    complement(2607..2706)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2723..2742)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2724..2742)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2747..2763)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2747..2763)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2865..2884)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2876..3685
                     /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(2943..2962)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3832..4420
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4321..4340
                     /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 accatgaggc caagcggcga tgaggacaga gcacggagaa ggcgccggag
      721 aaggcgccgg agagatctgc tgctgtccca gctgtgcttc ctggcctctg tggccctgct
      781 gctgtggagc ctgagctccc tgagggagca gaaggagctg gatctgatgg acctggtggg
      841 cgaggaccgg aagtggatga tggccagaaa gctgatgcaa gtgaatgata ccctgacaag
      901 cgaggacgca ggactgagga actccaagaa ttgcacagag ccagccctgc acgagttccc
      961 caacgatatc tttaccaatg aggacaggcg ccagggagca gtggtgctgc acgtgctgtg
     1021 cgccatctac atgttctatg ccctggccat cgtgtgcgac gatttctttg tgccaagcct
     1081 ggagaagatc tgcgagaggc tgcacctgtc cgaggatgtg gcaggagcca ccttcatggc
     1141 agcaggatct agcgcccccg agctgttcac ctccgtgatc ggcgtgttta tcacaaaggg
     1201 cgacgtggga gtgggaacca tcgtgggatc tgccgtgttc aatatcctgt gcatcatcgg
     1261 cgtgtgcgga ctgtttgcag gacaggtggt ggccctgtcc tcttggtgtc tgctgcggga
     1321 ttctatctac tataccctga gcgtgatcgc cctgatcgtg ttcatctacg acgagaaggt
     1381 gagctggtgg gagtccctgg tgctggtgct gatgtacctg atctatatcg tgatcatgaa
     1441 gtataacgcc tgcatccacc agtgttttga gcggagaaca aagggcgccg gcaacatggt
     1501 gaatggcctg gccaacaatg ccgagatcga cgatagctcc aactgcgatg ccaccgtggt
     1561 gctgctgaag aaggccaatt tccacaggaa ggcctccgtg atcatggtgg acgagctgct
     1621 gtctgcctac cctcaccagc tgagcttctc cgaggcagga ctgcgcatca tgatcacctc
     1681 ccactttccc cctaagacaa ggctgtccat ggcctctcgc atgctgatca acgagcggca
     1741 gagactgatc aactctcggg cctacaccaa tggcgagagc gaggtggcca tcaagatccc
     1801 aatcaagcac accgtggaga acggcacagg cccttctagc gccccagata gaggcgtgaa
     1861 tggcacaagg agggacgatg tggtggcaga ggcaggaaac gagaccgaga acgagaatga
     1921 ggataacgag aatgacgagg aggaggagga ggatgaggac gatgacgagg gcccatacac
     1981 ccccttcgac acaccttctg gcaagctgga gaccgtgaag tgggccttta catggccact
     2041 gagcttcgtg ctgtacttca ccgtgcccaa ctgtaataag cctcggtggg agaagtggtt
     2101 catggtgacc tttgcctcct ctacactgtg gatcgccgcc ttcagctaca tgatggtgtg
     2161 gatggtgacc atcatcggct atacactggg catccccgat gtgatcatgg gcatcacctt
     2221 tctggcagca ggcacatccg tgcctgactg catggcctct ctgatcgtgg ccagacaggg
     2281 catgggcgat atggccgtgt ctaacagcat cggctccaac gtgttcgaca tcctgatcgg
     2341 actgggactg ccatgggcac tgcagacact ggccgtggac tacggctcct atatcaggct
     2401 gaattctcgc ggcctgatct acagcgtggg actgctgctg gcaagcgtgt tcgtgaccgt
     2461 gtttggcgtg cacctgaaca agtggcagct ggataagaag ctgggctgcg gctgtctgct
     2521 gctgtatggc gtgttcctgt gcttttccat catgaccgag tttaacgtgt tcacatttgt
     2581 gaatctgcct atgtgcggcg accacgaccc agctttcttg tacaaagttg gcattataag
     2641 aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat
     2701 tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc
     2761 ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
     2821 atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag
     2881 ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata
     2941 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta
     3001 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     3061 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     3121 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa
     3181 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     3241 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     3301 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     3361 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     3421 acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     3481 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     3541 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     3601 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     3661 tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag
     3721 cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta
     3781 cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3841 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3901 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3961 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4021 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4081 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4141 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4201 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4261 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4321 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4381 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4441 tttacggttc ctggcctttt gctggccttt tgctcacatg tt
//
