LOCUS       sequence-357011-        5808 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 5808)
  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 5808)
  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_STOP
FEATURES             Location/Qualifiers
     source          1..5808
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3365..3464)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3481..3500)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3482..3500)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3505..3521)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3505..3521)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3623..3642)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3634..4443
                     /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(3701..3720)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4590..5178
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5079..5098
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5332..5349
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5508..5535)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5627..5713)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5762..5784
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5776..5793
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5777..5793
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacacat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccaccatgcc acacttcacc
      121 gtggtgccag tggacggacc taggagaggc gactacgata acctggaggg cctgtcttgg
      181 gtggactatg gcgagagagc cgagctggac gatagcgatg gccacggcaa tcacagggag
      241 agctccccat tcctgtcccc actggaggca tctaggggaa tcgactacta tgatcggaac
      301 ctggccctgt ttgaggagga gctggatatc cgccctaagg tgtctagcct gctgggcaag
      361 ctggtgtcct acaccaatct gacacaggga gcaaaggagc acgaggaggc agagtccgga
      421 gagggaacca ggcgccgggc agcagaggcc ccttctatgg gcacactgat gggcgtgtat
      481 ctgccatgcc tgcagaacat cttcggcgtg atcctgtttc tgaggctgac ctggatggtg
      541 ggaacagcag gcgtgctgca ggccctgctg atcgtgctga tctgctgttg ctgtaccctg
      601 ctgacagcca tctctatgag cgccatcgca accaatggag tggtgccagc aggaggaagc
      661 tacttcatga tctcccggtc tctgggacca gagtttggag gagcagtggg actgtgcttc
      721 tacctgggca ccacatttgc cgccgccatg tatatcctgg gcgccatcga gatcctgctg
      781 acctacatcg caccacctgc agccatcttc tatccctccg gcgcccacga cacctctaat
      841 gccacactga acaatatgag ggtgtacggc acaatcttcc tgacctttat gacactggtg
      901 gtgtttgtgg gcgtgaagta tgtgaacaag ttcgcctccc tgtttctggc ctgcgtgatc
      961 atctctatcc tgagcatcta cgccggcggc atcaagagca tcttcgaccc acccgtgttt
     1021 cccgtgtgca tgctgggcaa tcgcacgctt tctcgcgacc aatttgatat ttgtgccaag
     1081 acagccgtgg tggataacga gaccgtggcc acacagctgt ggagcttctt ttgccactcc
     1141 cccaatctga ccacagacag ctgtgatcct tacttcatgc tgaacaatgt gaccgagatc
     1201 cctggaatcc caggagcagc cgccggcgtg ctgcaggaga acctgtggag cgcctacctg
     1261 gagaagggcg acatcgtgga gaagcacgga ctgccatccg ccgatgcacc aagcctgaag
     1321 gagtccctgc cactgtatgt ggtggccgac atcgccacct ccttcacagt gctggtgggc
     1381 atcttctttc cttctgtgac cggcatcatg gcaggatcca atcgctctgg cgacctgcgg
     1441 gatgcccaga agagcatccc cgtgggcaca atcctggcca tcatcaccac atccctggtg
     1501 tacttctcct ctgtggtgct gtttggagca tgcatcgagg gagtggtgct gagggacaag
     1561 tatggcgatg gcgtgagcag gaacctggtg gtgggcaccc tggcatggcc atccccatgg
     1621 gtcatcgtga tcggcagctt cttttccacc tgtggagcag gactgcagag cctgacagga
     1681 gcaccaaggc tgctgcaggc aatcgcaaag gacaacatca tccctttcct gcgggtgttt
     1741 ggacacggca aggtcaatgg cgagccaacc tgggcactgc tgctgacagc cctgatcgca
     1801 gagctgggca tcctgatcgc cagcctggat atggtggccc ccatcctgtc catgttcttt
     1861 ctgatgtgct acctgttcgt gaacctggcc tgtgccgtgc agaccctgct gagaacacct
     1921 aattggcgcc cacggttcaa gtactatcac tgggccctgt cttttctggg catgagcctg
     1981 tgcctggccc tgatgtttgt gagctcctgg tactatgccc tggtggccat gctgatcgcc
     2041 ggcatgatct acaagtatat cgagtaccag ggagcagaga aggagtgggg cgacggaatc
     2101 aggggactga gcctgtccgc cgccagatat gcactgctga ggctggagga gggacctcca
     2161 cacaccaaga actggagacc tcagctgctg gtgctgctga agctggacga ggatctgcac
     2221 gtgaagtacc caaggctgct gaccttcgcc tctcagctga aggccggcaa gggcctgaca
     2281 atcgtgggca gcgtgatcca gggctctttt ctggagagct atggagaggc acaggcagca
     2341 gagcagacca tcaagaatat gatggagatc gagaaggtga agggcttctg ccaggtggtg
     2401 gtggcctcta aggtgagaga gggcctggcc cacctgatcc agtcttgtgg cctgggcggc
     2461 atgaggcaca acagcgtggt gctgggatgg ccatacggat ggaggcagtc cgaggaccca
     2521 agggcctgga agaccttcat cgatacagtg agatgcacca cagcagcaca cctggccctg
     2581 ctggtgccta agaacatcgc cttttaccca agcaatcacg agaggtatct ggagggccac
     2641 atcgacgtgt ggtggatcgt gcacgatggc ggcatgctga tgctgctgcc cttcctgctg
     2701 agacagcaca aagtgtggag gaagtgtaga atgaggatct ttaccgtggc ccagatggac
     2761 gataattcca tccagatgaa gaaggacctg gccgtgttcc tgtaccacct gagactggag
     2821 gccgaggtgg aggtggtgga gatgcacaac tctgatatca gcgcctacac ctatgagaga
     2881 acactgatga tggagcagag gagccagatg ctgagacaga tgaggctgac caagacagag
     2941 agggagaggg aggcacagct ggtgaaggac aggcactccg ccctgaggct ggagtccctg
     3001 tactctgacg aggaggatga gtctgccgtg ggcgccgaca agatccagat gacctggaca
     3061 cgcgataagt atatgaccga gacatgggac ccctctcacg cccctgataa tttccgggag
     3121 ctggtgcaca tcaagcctga tcagagcaac gtgagaagga tgcacaccgc cgtgaagctg
     3181 aatgaagtga tcgtgacaag atcccacgac gccaggctgg tgctgctgaa catgccagga
     3241 ccacctagga actctgaggg cgatgagaat tacatggagt ttctggaggt gctgaccgag
     3301 ggactggagc gggtgctgct ggtgagagga ggaggcagag aagtgatcac aatctatagc
     3361 taagacccag ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt
     3421 tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgatatc
     3481 ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct
     3541 caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg
     3601 tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtcg
     3661 aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat
     3721 aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag
     3781 ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga
     3841 ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct
     3901 gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca ggtattagaa
     3961 gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg
     4021 cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag
     4081 gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat
     4141 ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat
     4201 tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta
     4261 ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc
     4321 ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat
     4381 ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc
     4441 taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac ttgacgggac
     4501 ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca
     4561 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4621 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4681 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     4741 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4801 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4861 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4921 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4981 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5041 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5101 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5161 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     5221 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     5281 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     5341 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     5401 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     5461 gcaattaata cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa aggccatccg
     5521 tcaggatggc cttctgctta gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc
     5581 accctccggg ccgttgcttc acaacgttca aatccgctcc cggcggattt gtcctactca
     5641 ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtcttcc gactgagcct
     5701 ttcgttttat ttgatgcctg gcagttccct actctcgcgt taacgctagc atggatgttt
     5761 tcccagtcac gacgttgtaa aacgacggcc agtcttaagc tcgggccc
//
