LOCUS       sequence-259272-        6186 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 6186)
  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 6186)
  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_SLC12A2
FEATURES             Location/Qualifiers
     source          1..6186
                     /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(4311..4410)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4427..4446)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4428..4446)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4451..4467)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4451..4467)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4569..4588)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4580..5389
                     /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(4647..4666)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5536..6124
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6025..6044
                     /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 accatggagc cacggcccac agccccaagc tccggagcac caggactggc
      721 aggagtggga gagaccccat ccgccgccgc cctggccgcc gccagagtgg agctgcccgg
      781 cacagccgtg cccagcgtgc ctgaggacgc cgcccctgcc tccagggacg gcggcggcgt
      841 gcgcgatgag ggccctgccg ccgccggcga cggcctgggc aggccactgg gacctacccc
      901 aagccagtcc cgcttccagg tggatctggt gtccgagaac gccggcagag ccgccgcagc
      961 cgccgcagca gcagcagcag ccgccgccgc cgctggagcc ggcgccggcg ccaagcagac
     1021 ccccgccgac ggcgaggcat ctggagagag cgagcctgca aagggaagcg aggaggcaaa
     1081 gggccggttc agagtgaatt ttgtggatcc tgccgcctct agctccgccg aggactctct
     1141 gagcgatgca gcaggagtgg gagtggacgg accaaacgtg tccttccaga atggcggcga
     1201 tacagtgctg tctgagggct ctagcctgca ctctggcgga ggaggaggaa gcggacacca
     1261 ccagcactac tattacgaca cccacacaaa cacctattac ctgcggacat ttggccacaa
     1321 taccatggac gccgtgcctc ggatcgatca ctacagacac accgcagcac agctgggaga
     1381 gaagctgctg agaccaagcc tggccgagct gcacgacgag ctggagaagg agcccttcga
     1441 ggatggcttt gccaacggcg aggagtccac acctaccagg gatgccgtgg tgacatatac
     1501 cgccgagtct aagggcgtgg tgaagttcgg ctggatcaag ggcgtgctgg tgcggtgcat
     1561 gctgaatatc tggggcgtga tgctgtttat cagactgagc tggatcgtgg gacaggcagg
     1621 aatcggactg agcgtgctgg tcatcatgat ggccacagtg gtgaccacaa tcacaggcct
     1681 gtccacctct gccatcgcaa ccaacggatt cgtgcgggga ggaggcgcct attacctgat
     1741 cagcagatcc ctgggaccag agtttggagg agcaatcggc ctgatcttcg cctttgccaa
     1801 cgcagtggca gtggcaatgt acgtggtggg cttcgccgag acagtggtgg agctgctgaa
     1861 ggagcactcc atcctgatga tcgacgagat caatgatatc aggatcatcg gcgccatcac
     1921 cgtggtcatc ctgctgggca tcagcgtggc aggaatggag tgggaggcaa aggcccagat
     1981 cgtgctgctg gtcatcctgc tgctggccat cggcgacttt gtgatcggca ccttcatccc
     2041 actggagtct aagaagccca agggcttctt tggctacaag agcgagatct tcaacgagaa
     2101 ttttggcccc gatttccgcg aggaggagac attctttagc gtgttcgcca tcttctttcc
     2161 tgcagcaacc ggaatcctgg caggagcaaa catctccggc gacctggccg atcctcagtc
     2221 tgccatccca aagggcacac tgctggccat cctgatcacc acactggtgt acgtgggcat
     2281 cgccgtgtcc gtgggatctt gcgtggtgcg ggacgcaacc ggaaacgtga atgatacaat
     2341 cgtgaccgag ctgacaaact gcaccagcgc cgcctgtaag ctgaatttcg acttttcctc
     2401 ttgcgagagc tccccatgtt cctatggcct gatgaacaat tttcaggtca tgagcatggt
     2461 gtccggcttc acacccctga tcagcgccgg catcttttcc gccaccctgt ctagcgccct
     2521 ggcatctctg gtgagcgccc caaagatctt ccaggccctg tgcaaggata acatctatcc
     2581 cgccttccag atgtttgcca agggctacgg caagaacaat gagccactga gaggctatat
     2641 cctgaccttt ctgatcgccc tgggcttcat cctgatcgcc gagctgaacg tgatcgcccc
     2701 catcatctcc aatttctttc tggcctctta cgccctgatc aatttctccg tgtttcacgc
     2761 aagcctggca aagtcccctg gatggaggcc agccttcaag tattacaaca tgtggatcag
     2821 cctgctgggc gccatcctgt gctgtatcgt gatgtttgtg atcaattggt gggccgccct
     2881 gctgacatac gtgatcgtgc tgggcctgta tatctacgtg acctataaga agcctgacgt
     2941 gaactggggc tcctctacac aggccctgac ctatctgaat gccctgcagc actctatcag
     3001 gctgagcggc gtggaggatc acgtgaagaa cttccgccca cagtgcctgg tcatgacagg
     3061 agcaccaaat tccaggcctg ccctgctgca cctggtgcac gactttacca agaacgtggg
     3121 cctgatgatc tgtggacacg tgcacatggg accacggaga caggccatga aggagatgag
     3181 catcgatcag gccaagtacc agcggtggct gatcaagaat aagatgaagg ccttctatgc
     3241 accagtgcac gcagacgatc tgagggaggg cgcccagtac ctgatgcagg cagcaggact
     3301 gggaaggatg aagcctaaca ccctggtgct gggctttaag aaggactggc tgcaggccga
     3361 tatgagggac gtggatatgt acatcaatct gtttcacgac gccttcgata tccagtatgg
     3421 cgtggtggtc atccgcctga aggagggcct ggacatcagc cacctgcagg gccaggagga
     3481 gctgctgagc tcccaggaga agtctccagg caccaaggat gtggtggtga gcgtggagta
     3541 ctccaagaag tctgacctgg atacatccaa gcccctgtct gagaagccta tcacccacaa
     3601 ggtggaggag gaggacggca agacagccac ccagcccctg ctgaagaagg agtctaaggg
     3661 cccaatcgtg cccctgaacg tggccgacca gaagctgctg gaggccagca cacagttcca
     3721 gaagaagcag ggcaagaata ccatcgacgt gtggtggctg tttgacgatg gcggcctgac
     3781 cctgctgatc ccctacctgc tgaccaccaa gaagaagtgg aaggactgta agatccgggt
     3841 gttcatcggc ggcaagatca acagaatcga ccacgatagg cgcgccatgg ccacactgct
     3901 gtctaagttt agaatcgact tcagcgatat catggtgctg ggcgatatca acaccaagcc
     3961 taagaaggag aatatcatcg cctttgagga gatcatcgag ccatatcggc tgcacgagga
     4021 cgataaggag caggacatcg ccgataagat gaaggaggac gagccctgga gaatcacaga
     4081 taacgagctg gagctgtata agacaaagac ctacaggcag atccgcctga acgaactgct
     4141 gaaggaacac tctagcaccg ccaatatcat cgtgatgtcc ctgcctgtgg caaggaaggg
     4201 agccgtgtcc tctgccctgt acatggcctg gctggaggcc ctgtctaagg acctgccacc
     4261 tatcctgctg gtgcgcggca atcaccagag cgtgctgacc ttctattccg acccagcttt
     4321 cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt
     4381 cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc
     4441 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     4501 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     4561 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     4621 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     4681 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     4741 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     4801 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     4861 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     4921 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     4981 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     5041 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     5101 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     5161 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     5221 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     5281 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     5341 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     5401 taattggttg taacactggc agagcattac gctgacttga cgggacggcg caagctcatg
     5461 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     5521 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5581 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5641 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5701 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5761 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5821 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5881 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5941 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6001 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6061 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6121 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6181 catgtt
//
