LOCUS       sequence-345629-        5616 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 5616)
  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 5616)
  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-SLC12A3_STOP
FEATURES             Location/Qualifiers
     source          1..5616
                     /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(3173..3272)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3289..3308)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3290..3308)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3313..3329)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3313..3329)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3431..3450)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3442..4251
                     /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(3509..3528)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4398..4986
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4887..4906
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5140..5157
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5316..5343)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5435..5521)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5570..5592
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5584..5601
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5585..5601
                     /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 ccaccatggc agagctgcca
      121 accacagaga cccctggcga cgccacactg tgctctggcc ggtttaccat cagcacactg
      181 ctgagctccg acgagccatc tccacctgca gcctacgatt ctagccaccc tagccacctg
      241 acccactcct ctacattctg tatgagaacc tttggctata acacaatcga cgtggtgcca
      301 acctacgagc actatgccaa ttccacacag ccaggagagc caaggaaggt gagacccacc
      361 ctggccgatc tgcactcttt tctgaagcag gagggcaggc acctgcacgc actggccttc
      421 gactctaggc ctagccacga gatgacagat ggactggtgg agggagaggc aggcaccagc
      481 tccgagaaga accctgagga gccagtgcgg ttcggctggg tgaagggcgt gatgatcaga
      541 tgcatgctga atatctgggg cgtgatcctg tacctgaggc tgccatggat caccgcacag
      601 gcaggcatcg tgctgacatg gatcatcatc ctgctgtccg tgaccgtgac atctatcacc
      661 ggcctgtcta tcagcgccat cagcacaaac ggcaaggtga agtccggcgg cacctacttt
      721 ctgatctccc gctctctggg accagagctg ggaggctcca tcggcctgat cttcgccttt
      781 gccaacgcag tgggagtggc aatgcacacc gtgggattcg ccgagacagt gagggatctg
      841 ctgcaggagt atggcgcccc catcgtggac cctatcaatg atatccgcat catcgccgtg
      901 gtgtctgtga cagtgctgct ggccatcagc ctggccggca tggagtggga gtccaaggcc
      961 caggtgctgt tcttcctggt catcatggtg agcttcgcca actacctggt gggcaccctg
     1021 atcccaccca gcgaggacaa ggcctccaag ggcttctttt cttatagggc cgatatcttt
     1081 gtgcagaatc tggtgccaga ctggagggga ccagatggca cattctttgg catgttcagc
     1141 atcttctttc catccgccac cggaatcctg gcaggagcaa acatctccgg cgacctgaag
     1201 gatcctgcca tcgccatccc aaagggcaca ctgatggcca tcttttggac cacaatctct
     1261 tacctggcca tcagcgccac catcggatcc tgcgtggtgc gggacgcaag cggcgtgctg
     1321 aatgatactg tgacaccagg atggggagca tgcgagggac tggcatgtag ctacggctgg
     1381 aacttcaccg agtgcacaca gcagcactcc tgtcactatg gcctgatcaa ttactatcag
     1441 accatgtcca tggtgtctgg attcgcacca ctgatcaccg caggaatctt tggcgccaca
     1501 ctgtctagcg ccctggcatg tctggtgagc gccgcaaagg tgtttcagtg cctgtgcgag
     1561 gaccagctgt accctctgat cggcttcttt ggcaagggct atggcaagaa caaggagcca
     1621 gtgcggggct acctgctggc ctatgccatc gccgtggcct tcatcatcat cgccgagctg
     1681 aacacaatcg cccccatcat ctccaatttc tttctgtgct cttacgccct gatcaacttc
     1741 tcttgttttc acgcctctat caccaatagc ccaggctgga gacccagctt ccagtactat
     1801 aataagtggg ccgccctgtt tggcgccatc atctccgtgg tcatcatgtt cctgctgacc
     1861 tggtgggccg ccctgatcgc aatcggagtg gtgctgtttc tgctgctgta cgtgatctat
     1921 aagaagcctg aggtgaactg gggctcctct gtgcaggccg gcagctacaa tctggccctg
     1981 agctattccg tgggcctgaa cgaggtggag gaccacatca agaattacag accacagtgc
     2041 ctggtgctga caggacctcc aaacttccgg cccgccctgg tggatttcgt gggcaccttt
     2101 acaagaaatc tgtccctgat gatctgtggc cacgtgctga tcggccccca caagcagcgg
     2161 atgcctgagc tgcagctgat cgccaacggc cacaccaagt ggctgaataa gagaaagatc
     2221 aaggcctttt atagcgacgt gatcgccgag gatctgcgga gaggagtgca gatcctgatg
     2281 caggcagcag gactgggaag gatgaagccc aacatcctgg tggtgggctt caagaagaat
     2341 tggcagtccg cccaccctgc aaccgtggag gactacatcg gcatcctgca cgacgccttc
     2401 gacttcaact atggcgtgtg cgtgatgagg atgcgcgagg gcctgaacgt gtctaagatg
     2461 atgcaggccc acatcaatcc cgtgttcgac ccagccgagg atggcaagga ggcaagcgcc
     2521 agggtggacc caaaggccct ggtgaaggag gagcaggcca ccacaatctt tcagtccgag
     2581 cagggcaaga agacaatcga tatctactgg ctgttcgacg atggcggcct gaccctgctg
     2641 atcccttatc tgctgggcag gaagaggcgc tggagcaagt gcaagatccg cgtgtttgtg
     2701 ggcggccaga tcaaccggat ggatcaggag agaaaggcca tcatcagcct gctgtccaag
     2761 ttccggctgg gctttcacga ggtgcacatc ctgccagaca tcaaccagaa tcccagggcc
     2821 gagcacacca agcggttcga ggatatgatc gcccctttca ggctgaacga cggctttaag
     2881 gatgaggcca cagtgaatga gatgcggaga gactgtccat ggaagatctc cgatgaggag
     2941 atcaccaaga accgcgtgaa gtctctgagg caggtgcgcc tgaatgagat cgtgctggac
     3001 tactctcggg atgccgccct gatcgtgatc acactgccta tcggcagaaa gggcaagtgc
     3061 ccaagctccc tgtatatggc ctggctggag accctgagcc aggacctgag gccacctgtg
     3121 atcctgatca gaggcaacca ggagaatgtg ctgaccttct actgtcagta agacccagct
     3181 ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag
     3241 gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc ctatagtgag
     3301 tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga
     3361 tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata
     3421 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta
     3481 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
     3541 tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa
     3601 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
     3661 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
     3721 ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat
     3781 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
     3841 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
     3901 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
     3961 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact
     4021 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
     4081 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     4141 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     4201 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg
     4261 gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca
     4321 tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa
     4381 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4441 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4501 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     4561 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     4621 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     4681 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     4741 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     4801 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     4861 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     4921 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     4981 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     5041 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     5101 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     5161 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     5221 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg
     5281 cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct
     5341 tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc
     5401 gttgcttcac aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca
     5461 ccgacaaaca acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt
     5521 gatgcctggc agttccctac tctcgcgtta acgctagcat ggatgttttc ccagtcacga
     5581 cgttgtaaaa cgacggccag tcttaagctc gggccc
//
