LOCUS       sequence-259333-        5613 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 5613)
  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 5613)
  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
FEATURES             Location/Qualifiers
     source          1..5613
                     /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(3738..3837)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3854..3873)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3855..3873)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3878..3894)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3878..3894)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3996..4015)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4007..4816
                     /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(4074..4093)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4963..5551
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5452..5471
                     /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 accatggcag agctgccaac cacagagacc cctggcgacg ccacactgtg
      721 ctctggccgg tttaccatca gcacactgct gagctccgac gagccatctc cacctgcagc
      781 ctacgattct agccacccta gccacctgac ccactcctct acattctgta tgagaacctt
      841 tggctataac acaatcgacg tggtgccaac ctacgagcac tatgccaatt ccacacagcc
      901 aggagagcca aggaaggtga gacccaccct ggccgatctg cactcttttc tgaagcagga
      961 gggcaggcac ctgcacgcac tggccttcga ctctaggcct agccacgaga tgacagatgg
     1021 actggtggag ggagaggcag gcaccagctc cgagaagaac cctgaggagc cagtgcggtt
     1081 cggctgggtg aagggcgtga tgatcagatg catgctgaat atctggggcg tgatcctgta
     1141 cctgaggctg ccatggatca ccgcacaggc aggcatcgtg ctgacatgga tcatcatcct
     1201 gctgtccgtg accgtgacat ctatcaccgg cctgtctatc agcgccatca gcacaaacgg
     1261 caaggtgaag tccggcggca cctactttct gatctcccgc tctctgggac cagagctggg
     1321 aggctccatc ggcctgatct tcgcctttgc caacgcagtg ggagtggcaa tgcacaccgt
     1381 gggattcgcc gagacagtga gggatctgct gcaggagtat ggcgccccca tcgtggaccc
     1441 tatcaatgat atccgcatca tcgccgtggt gtctgtgaca gtgctgctgg ccatcagcct
     1501 ggccggcatg gagtgggagt ccaaggccca ggtgctgttc ttcctggtca tcatggtgag
     1561 cttcgccaac tacctggtgg gcaccctgat cccacccagc gaggacaagg cctccaaggg
     1621 cttcttttct tatagggccg atatctttgt gcagaatctg gtgccagact ggaggggacc
     1681 agatggcaca ttctttggca tgttcagcat cttctttcca tccgccaccg gaatcctggc
     1741 aggagcaaac atctccggcg acctgaagga tcctgccatc gccatcccaa agggcacact
     1801 gatggccatc ttttggacca caatctctta cctggccatc agcgccacca tcggatcctg
     1861 cgtggtgcgg gacgcaagcg gcgtgctgaa tgatactgtg acaccaggat ggggagcatg
     1921 cgagggactg gcatgtagct acggctggaa cttcaccgag tgcacacagc agcactcctg
     1981 tcactatggc ctgatcaatt actatcagac catgtccatg gtgtctggat tcgcaccact
     2041 gatcaccgca ggaatctttg gcgccacact gtctagcgcc ctggcatgtc tggtgagcgc
     2101 cgcaaaggtg tttcagtgcc tgtgcgagga ccagctgtac cctctgatcg gcttctttgg
     2161 caagggctat ggcaagaaca aggagccagt gcggggctac ctgctggcct atgccatcgc
     2221 cgtggccttc atcatcatcg ccgagctgaa cacaatcgcc cccatcatct ccaatttctt
     2281 tctgtgctct tacgccctga tcaacttctc ttgttttcac gcctctatca ccaatagccc
     2341 aggctggaga cccagcttcc agtactataa taagtgggcc gccctgtttg gcgccatcat
     2401 ctccgtggtc atcatgttcc tgctgacctg gtgggccgcc ctgatcgcaa tcggagtggt
     2461 gctgtttctg ctgctgtacg tgatctataa gaagcctgag gtgaactggg gctcctctgt
     2521 gcaggccggc agctacaatc tggccctgag ctattccgtg ggcctgaacg aggtggagga
     2581 ccacatcaag aattacagac cacagtgcct ggtgctgaca ggacctccaa acttccggcc
     2641 cgccctggtg gatttcgtgg gcacctttac aagaaatctg tccctgatga tctgtggcca
     2701 cgtgctgatc ggcccccaca agcagcggat gcctgagctg cagctgatcg ccaacggcca
     2761 caccaagtgg ctgaataaga gaaagatcaa ggccttttat agcgacgtga tcgccgagga
     2821 tctgcggaga ggagtgcaga tcctgatgca ggcagcagga ctgggaagga tgaagcccaa
     2881 catcctggtg gtgggcttca agaagaattg gcagtccgcc caccctgcaa ccgtggagga
     2941 ctacatcggc atcctgcacg acgccttcga cttcaactat ggcgtgtgcg tgatgaggat
     3001 gcgcgagggc ctgaacgtgt ctaagatgat gcaggcccac atcaatcccg tgttcgaccc
     3061 agccgaggat ggcaaggagg caagcgccag ggtggaccca aaggccctgg tgaaggagga
     3121 gcaggccacc acaatctttc agtccgagca gggcaagaag acaatcgata tctactggct
     3181 gttcgacgat ggcggcctga ccctgctgat cccttatctg ctgggcagga agaggcgctg
     3241 gagcaagtgc aagatccgcg tgtttgtggg cggccagatc aaccggatgg atcaggagag
     3301 aaaggccatc atcagcctgc tgtccaagtt ccggctgggc tttcacgagg tgcacatcct
     3361 gccagacatc aaccagaatc ccagggccga gcacaccaag cggttcgagg atatgatcgc
     3421 ccctttcagg ctgaacgacg gctttaagga tgaggccaca gtgaatgaga tgcggagaga
     3481 ctgtccatgg aagatctccg atgaggagat caccaagaac cgcgtgaagt ctctgaggca
     3541 ggtgcgcctg aatgagatcg tgctggacta ctctcgggat gccgccctga tcgtgatcac
     3601 actgcctatc ggcagaaagg gcaagtgccc aagctccctg tatatggcct ggctggagac
     3661 cctgagccag gacctgaggc cacctgtgat cctgatcaga ggcaaccagg agaatgtgct
     3721 gaccttctac tgtcaggacc cagctttctt gtacaaagtt ggcattataa gaaagcattg
     3781 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     3841 tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     3901 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     3961 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     4021 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     4081 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     4141 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     4201 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     4261 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
     4321 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     4381 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     4441 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     4501 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     4561 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     4621 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     4681 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     4741 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     4801 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     4861 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     4921 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     4981 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5041 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5101 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5161 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5221 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5281 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5341 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5401 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     5461 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5521 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5581 cctggccttt tgctggcctt ttgctcacat gtt
//
