LOCUS       sequence-259495-        5805 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 5805)
  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 5805)
  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_SLC12A4
FEATURES             Location/Qualifiers
     source          1..5805
                     /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(3930..4029)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4046..4065)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4047..4065)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4070..4086)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4070..4086)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4188..4207)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4199..5008
                     /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(4266..4285)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5155..5743
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5644..5663
                     /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 accatgccac acttcaccgt ggtgccagtg gacggaccta ggagaggcga
      721 ctacgataac ctggagggcc tgtcttgggt ggactatggc gagagagccg agctggacga
      781 tagcgatggc cacggcaatc acagggagag ctccccattc ctgtccccac tggaggcatc
      841 taggggaatc gactactatg atcggaacct ggccctgttt gaggaggagc tggatatccg
      901 ccctaaggtg tctagcctgc tgggcaagct ggtgtcctac accaatctga cacagggagc
      961 aaaggagcac gaggaggcag agtccggaga gggaaccagg cgccgggcag cagaggcccc
     1021 ttctatgggc acactgatgg gcgtgtatct gccatgcctg cagaacatct tcggcgtgat
     1081 cctgtttctg aggctgacct ggatggtggg aacagcaggc gtgctgcagg ccctgctgat
     1141 cgtgctgatc tgctgttgct gtaccctgct gacagccatc tctatgagcg ccatcgcaac
     1201 caatggagtg gtgccagcag gaggaagcta cttcatgatc tcccggtctc tgggaccaga
     1261 gtttggagga gcagtgggac tgtgcttcta cctgggcacc acatttgccg ccgccatgta
     1321 tatcctgggc gccatcgaga tcctgctgac ctacatcgca ccacctgcag ccatcttcta
     1381 tccctccggc gcccacgaca cctctaatgc cacactgaac aatatgaggg tgtacggcac
     1441 aatcttcctg acctttatga cactggtggt gtttgtgggc gtgaagtatg tgaacaagtt
     1501 cgcctccctg tttctggcct gcgtgatcat ctctatcctg agcatctacg ccggcggcat
     1561 caagagcatc ttcgacccac ccgtgtttcc cgtgtgcatg ctgggcaatc gcacgctttc
     1621 tcgcgaccaa tttgatattt gtgccaagac agccgtggtg gataacgaga ccgtggccac
     1681 acagctgtgg agcttctttt gccactcccc caatctgacc acagacagct gtgatcctta
     1741 cttcatgctg aacaatgtga ccgagatccc tggaatccca ggagcagccg ccggcgtgct
     1801 gcaggagaac ctgtggagcg cctacctgga gaagggcgac atcgtggaga agcacggact
     1861 gccatccgcc gatgcaccaa gcctgaagga gtccctgcca ctgtatgtgg tggccgacat
     1921 cgccacctcc ttcacagtgc tggtgggcat cttctttcct tctgtgaccg gcatcatggc
     1981 aggatccaat cgctctggcg acctgcggga tgcccagaag agcatccccg tgggcacaat
     2041 cctggccatc atcaccacat ccctggtgta cttctcctct gtggtgctgt ttggagcatg
     2101 catcgaggga gtggtgctga gggacaagta tggcgatggc gtgagcagga acctggtggt
     2161 gggcaccctg gcatggccat ccccatgggt catcgtgatc ggcagcttct tttccacctg
     2221 tggagcagga ctgcagagcc tgacaggagc accaaggctg ctgcaggcaa tcgcaaagga
     2281 caacatcatc cctttcctgc gggtgtttgg acacggcaag gtcaatggcg agccaacctg
     2341 ggcactgctg ctgacagccc tgatcgcaga gctgggcatc ctgatcgcca gcctggatat
     2401 ggtggccccc atcctgtcca tgttctttct gatgtgctac ctgttcgtga acctggcctg
     2461 tgccgtgcag accctgctga gaacacctaa ttggcgccca cggttcaagt actatcactg
     2521 ggccctgtct tttctgggca tgagcctgtg cctggccctg atgtttgtga gctcctggta
     2581 ctatgccctg gtggccatgc tgatcgccgg catgatctac aagtatatcg agtaccaggg
     2641 agcagagaag gagtggggcg acggaatcag gggactgagc ctgtccgccg ccagatatgc
     2701 actgctgagg ctggaggagg gacctccaca caccaagaac tggagacctc agctgctggt
     2761 gctgctgaag ctggacgagg atctgcacgt gaagtaccca aggctgctga ccttcgcctc
     2821 tcagctgaag gccggcaagg gcctgacaat cgtgggcagc gtgatccagg gctcttttct
     2881 ggagagctat ggagaggcac aggcagcaga gcagaccatc aagaatatga tggagatcga
     2941 gaaggtgaag ggcttctgcc aggtggtggt ggcctctaag gtgagagagg gcctggccca
     3001 cctgatccag tcttgtggcc tgggcggcat gaggcacaac agcgtggtgc tgggatggcc
     3061 atacggatgg aggcagtccg aggacccaag ggcctggaag accttcatcg atacagtgag
     3121 atgcaccaca gcagcacacc tggccctgct ggtgcctaag aacatcgcct tttacccaag
     3181 caatcacgag aggtatctgg agggccacat cgacgtgtgg tggatcgtgc acgatggcgg
     3241 catgctgatg ctgctgccct tcctgctgag acagcacaaa gtgtggagga agtgtagaat
     3301 gaggatcttt accgtggccc agatggacga taattccatc cagatgaaga aggacctggc
     3361 cgtgttcctg taccacctga gactggaggc cgaggtggag gtggtggaga tgcacaactc
     3421 tgatatcagc gcctacacct atgagagaac actgatgatg gagcagagga gccagatgct
     3481 gagacagatg aggctgacca agacagagag ggagagggag gcacagctgg tgaaggacag
     3541 gcactccgcc ctgaggctgg agtccctgta ctctgacgag gaggatgagt ctgccgtggg
     3601 cgccgacaag atccagatga cctggacacg cgataagtat atgaccgaga catgggaccc
     3661 ctctcacgcc cctgataatt tccgggagct ggtgcacatc aagcctgatc agagcaacgt
     3721 gagaaggatg cacaccgccg tgaagctgaa tgaagtgatc gtgacaagat cccacgacgc
     3781 caggctggtg ctgctgaaca tgccaggacc acctaggaac tctgagggcg atgagaatta
     3841 catggagttt ctggaggtgc tgaccgaggg actggagcgg gtgctgctgg tgagaggagg
     3901 aggcagagaa gtgatcacaa tctatagcga cccagctttc ttgtacaaag ttggcattat
     3961 aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat
     4021 cattatttgc catccagctg atatccccta tagtgagtcg tattacatgg tcatagctgt
     4081 ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa
     4141 tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat
     4201 gagccatatt caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt
     4261 atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt
     4321 gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa
     4381 tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac
     4441 catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg
     4501 aaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc
     4561 gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag
     4621 cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc
     4681 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca
     4741 taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa
     4801 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc
     4861 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt
     4921 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt
     4981 tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggca
     5041 gagcattacg ctgacttgac gggacggcgc aagctcatga ccaaaatccc ttaacgtgag
     5101 ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5161 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5221 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5281 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5341 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5401 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5461 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5521 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5581 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5641 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5701 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5761 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtt
//
