LOCUS       sequence-312154-        5970 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 5970)
  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 5970)
  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-SLC12A5_STOP
FEATURES             Location/Qualifiers
     source          1..5970
                     /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(4095..4194)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4211..4230)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4212..4230)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4235..4251)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4235..4251)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4353..4372)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4364..5173
                     /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(4431..4450)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5320..5908
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5809..5828
                     /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 accatgtctc ggagattcac cgtgacaagc ctgccacctg caggaccagc
      721 acgctctcca gacccagagt ccaggcgcca ctctgtggca gatccaaggc acctgccagg
      781 agaggacgtg aagggcgatg gcaacccaaa ggagagctcc cccttcatca atagcaccga
      841 cacagagaag ggcaaggagt atgatggcaa gaacatggcc ctgtttgagg aggagatgga
      901 cacatcccca atggtgtcta gcctgctgtc tggcctggcc aactacacca atctgccaca
      961 gggaagcaga gagcacgagg aggccgagaa caatgagggc ggcaagaaga agcctgtgca
     1021 ggccccaagg atgggcacat tcatgggcgt gtacctgcct tgcctgcaga acatcttcgg
     1081 cgtgatcctg tttctgaggc tgacctgggt ggtgggaatc gcaggcatca tggagagctt
     1141 ctgcatggtg tttatctgct gttcctgtac catgctgaca gccatctcta tgagcgccat
     1201 cgcaacaaat ggagtggtgc cagcaggagg atcctactat atgatctccc ggtctctggg
     1261 acctgagttc ggaggagcag tgggactgtg cttctatctg ggcaccacat ttgccggcgc
     1321 catgtacatc ctgggcacca tcgagatcct gctggcctat ctgttcccag ccatggccat
     1381 ctttaaggca gaggacgcaa gcggagaggc agcagccatg ctgaacaata tgagagtgta
     1441 tggcacctgc gtgctgacat gtatggccac cgtggtgttt gtgggcgtga agtacgtgaa
     1501 taagttcgcc ctggtgtttc tgggctgcgt gatcctgagc atcctggcca tctacgccgg
     1561 cgtgatcaag tccgccttcg acccacccaa ctttccaatc tgcctgctgg gcaatagaac
     1621 cctgtccagg cacggcttcg acgtgtgcgc aaagctggca tgggagggca acgagacagt
     1681 gaccacaaga ctgtggggcc tgttctgctc ctctaggttt ctgaatgcca cctgtgatga
     1741 gtacttcacc agaaacaatg tgaccgagat ccagggaatc ccaggagcag caagcggact
     1801 gatcaaggag aacctgtgga gctcctacct gacaaagggc gtgatcgtgg agaggagcgg
     1861 aatgacatcc gtgggactgg cagacggaac ccctatcgac atggatcacc catacgtgtt
     1921 ctctgatatg acaagctact tcaccctgct ggtgggcatc tacttcccat ccgtgaccgg
     1981 aatcatggca ggatctaata gaagcggcga cctgagggat gcccagaaga gcatccctac
     2041 cggcacaatc ctggccatcg ccaccacatc cgccgtgtat atctctagcg tggtgctgtt
     2101 cggagcatgc atcgagggag tggtgctgag agacaagttt ggcgaggccg tgaacggcaa
     2161 tctggtggtg ggcaccctgg catggcctag cccatgggtc atcgtgatcg gctccttctt
     2221 ttctacatgt ggagcaggac tgcagtccct gaccggagca cctaggctgc tgcaggcaat
     2281 ctctagggac ggaatcgtgc cattcctgca ggtgtttggc cacggcaagg ccaacggcga
     2341 gccaacatgg gccctgctgc tgaccgcctg catctgtgag atcggcatcc tgatcgcctc
     2401 tctggatgag gtggccccca tcctgagcat gttctttctg atgtgctaca tgttcgtgaa
     2461 cctggcctgt gccgtgcaga cactgctgag gacccccaat tggaggcctc gcttccggta
     2521 ctatcactgg accctgtcct ttctgggcat gtctctgtgc ctggccctga tgtttatctg
     2581 tagctggtac tatgcactgg tggcaatgct gatcgcagga ctgatctaca agtatatcga
     2641 gtatagaggc gccgagaagg agtggggcga tggcatcagg ggcctgagcc tgtccgccgc
     2701 aagatacgca ctgctgaggc tggaggaggg acctccacac acaaagaact ggaggcctca
     2761 gctgctggtg ctggtgcggg tggaccagga tcagaatgtg gtgcacccac agctgctgag
     2821 cctgacctcc cagctgaagg ccggcaaggg cctgacaatc gtgggctctg tgctggaggg
     2881 caccttcctg gagaaccacc cacaggcaca gagggcagag gagagcatcc ggagactgat
     2941 ggaggccgag aaggtgaagg gcttttgcca ggtggtcatc tcctctaatc tgcgggacgg
     3001 cgtgagccac ctgatccagt ccggaggact gggaggactg cagcacaaca cagtgctggt
     3061 gggctggcct cgcaattggc ggcagaagga ggatcaccag acctggagaa acttcatcga
     3121 gctggtgagg gagaccacag caggacacct ggccctgctg gtgaccaaga acgtgtccat
     3181 gttccctggc aatccagagc ggttcagcga gggcagcatc gacgtgtggt ggatcgtgca
     3241 cgatggcggc atgctgatgc tgctgccctt cctgctgcgg caccacaaag tgtggagaaa
     3301 gtgtaagatg aggatcttta cagtggccca gatggacgat aatagcatcc agatgaagaa
     3361 ggacctgacc acatttctgt accacctgcg catcaccgcc gaggtggagg tggtggagat
     3421 gcacgagtcc gatatctctg cctacacata tgagaagacc ctggtcatgg agcagagatc
     3481 ccagatcctg aagcagatgc acctgacaaa gaacgagaga gagagggaga tccagtctat
     3541 caccgacgag agcaggggct ccatcaggcg caagaaccca gccaatacac gcctgcggct
     3601 gaatgtgcca gaggagaccg caggcgacag cgaggagaag cctgaggagg aggtgcagct
     3661 gatccacgat cagtctgccc caagctgccc tagctcctct ccatcccctg gagaggagcc
     3721 agagggagag ggagagaccg accctgagaa ggtgcacctg acctggacaa aggataagtc
     3781 cgtggccgag aagaacaagg gcccatctcc cgtgagctcc gagggcatca aggacttctt
     3841 ttctatgaag cctgagtggg agaacctgaa tcagagcaac gtgcggagaa tgcacaccgc
     3901 cgtgcgcctg aatgaagtga tcgtgaagaa gtcccgggat gccaagctgg tgctgctgaa
     3961 catgccaggc ccccctagaa acaggaatgg cgacgagaat tatatggagt tcctggaggt
     4021 gctgacagag cacctggata gagtgatgct ggtgcgcgga ggaggaagag aagtgatcac
     4081 catctactct taagacccag ctttcttgta caaagttggc attataagaa agcattgctt
     4141 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     4201 agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
     4261 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaataatat catcatgaac
     4321 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
     4381 ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     4441 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga
     4501 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     4561 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     4621 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca
     4681 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     4741 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     4801 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     4861 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
     4921 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     4981 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     5041 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     5101 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     5161 tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac
     5221 ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     5281 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5341 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5401 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5461 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5521 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5581 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5641 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5701 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5761 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5821 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5881 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5941 ggccttttgc tggccttttg ctcacatgtt
//
