LOCUS       sequence-312222-        4485 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 4485)
  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 4485)
  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_STOP
FEATURES             Location/Qualifiers
     source          1..4485
                     /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(2610..2709)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2726..2745)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2727..2745)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2750..2766)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2750..2766)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2868..2887)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2879..3688
                     /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(2946..2965)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3835..4423
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4324..4343
                     /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 accactaaga cccagctttc ttgtacaaag ttggcattat
     2641 aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat
     2701 cattatttgc catccagctg atatccccta tagtgagtcg tattacatgg tcatagctgt
     2761 ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaat
     2821 aatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat
     2881 gagccatatt caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt
     2941 atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt
     3001 gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa
     3061 tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac
     3121 catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg
     3181 aaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc
     3241 gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag
     3301 cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc
     3361 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca
     3421 taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa
     3481 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc
     3541 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt
     3601 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt
     3661 tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggca
     3721 gagcattacg ctgacttgac gggacggcgc aagctcatga ccaaaatccc ttaacgtgag
     3781 ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3841 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3901 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3961 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     4021 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4081 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4141 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4201 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4261 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4321 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4381 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4441 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtt
//
