LOCUS       sequence-259223-        4293 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 4293)
  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 4293)
  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_SLC15A3
FEATURES             Location/Qualifiers
     source          1..4293
                     /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(2418..2517)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2534..2553)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2535..2553)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2558..2574)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2558..2574)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2676..2695)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2687..3496
                     /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(2754..2773)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3643..4231
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4132..4151
                     /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 accatgccag cccccagggc ccgcgagcag ccaagggtgc ccggcgagcg
      721 ccagcctctg ctgccaaggg gcgcccgcgg cccacggaga tggaggaggg cagcaggagc
      781 agccgtgctg ctggtggaga tgctggagag ggcagccttc tttggcgtga cagccaacct
      841 ggtgctgtac ctgaattcta caaatttcaa ttggacaggc gaacaggcaa ccagagccgc
      901 cctggtgttt ctgggagcaa gctatctgct ggcacctgtg ggaggatggc tggcagacgt
      961 gtacctgggc cggtatagag ccgtggccct gtctctgctg ctgtacctgg cagcaagcgg
     1021 actgctgcca gcaaccgcct tccctgacgg caggagctcc ttttgcggcg agatgccagc
     1081 atcccctctg ggaccagcat gcccatctgc cggatgtcct cgctctagcc ctagcccata
     1141 ctgtgcccca gtgctgtatg caggactgct gctgctggga ctggcagcat cctctgtgcg
     1201 gtctaacctg acaagcttcg gcgccgacca ggtcatggat ctgggcagag acgccacccg
     1261 gagattcttt aactggtttt actggagcat caatctggga gccgtgctgt ccctgctggt
     1321 ggtggccttc atccagcaga atatctcctt tctgctgggc tattctatcc cagtgggctg
     1381 cgtgggcctg gccttcttta tcttcctgtt tgccacaccc gtgttcatca ccaagccccc
     1441 tatgggctcc caggtgagct ccatgctgaa gctggccctg cagaactgct gtccccagct
     1501 gtggcagagg cactctgcca gggataggca gtgtgcacgg gtgctggcag acgagagaag
     1561 cccacagcct ggagcatccc ctcaggagga tatcgccaat tttcaggtgc tggtgaagat
     1621 cctgccagtg atggtgacac tggtgcccta ctggatggtg tatttccaga tgcagagcac
     1681 ctacgtgctg cagggcctgc acctgcacat ccctaacatc tttcctgcca acccagccaa
     1741 tatctccgtg gccctgaggg cacagggatc tagctataca atcccagagg cctggctgct
     1801 gctggccaat gtggtggtgg tgctgatcct ggtgcccctg aaggataggc tgatcgaccc
     1861 tctgctgctg aggtgcaagc tgctgccaag cgccctgcag aagatggcac tgggcatgtt
     1921 ctttggcttc acatccgtga tcgtggcagg cgtgctggag atggagcggc tgcactacat
     1981 ccaccacaac gagaccgtgt cccagcagat cggcgaggtg ctgtacaatg ccgccccact
     2041 gtctatctgg tggcagatcc cccagtatct gctgatcggc atcagcgaga tcttcgcctc
     2101 catccccggc ctggagtttg cctattctga ggcccctaga agcatgcagg gcgccatcat
     2161 gggcatcttc ttttgtctgt ccggagtggg atctctgctg ggatcctctc tggtggccct
     2221 gctgtccctg cctggaggat ggctgcactg cccaaaggat ttcggcaaca tcaacaattg
     2281 taggatggac ctgtacttct ttctgctggc aggaatccag gcagtgaccg ccctgctgtt
     2341 cgtgtggatc gcaggccggt atgagagagc atctcaggga ccagcaagcc actcccgctt
     2401 tagccgggac agaggcgacc cagctttctt gtacaaagtt ggcattataa gaaagcattg
     2461 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     2521 tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     2581 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     2641 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     2701 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     2761 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     2821 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     2881 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     2941 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
     3001 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     3061 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     3121 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     3181 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     3241 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     3301 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     3361 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     3421 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     3481 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     3541 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     3601 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3661 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3721 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3781 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3841 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3901 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3961 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     4021 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     4081 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     4141 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     4201 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     4261 cctggccttt tgctggcctt ttgctcacat gtt
//
