LOCUS       sequence-259447-        4287 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 4287)
  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 4287)
  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_SLC15A5
FEATURES             Location/Qualifiers
     source          1..4287
                     /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(2412..2511)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2528..2547)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2529..2547)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2552..2568)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2552..2568)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2670..2689)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2681..3490
                     /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(2748..2767)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3637..4225
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4126..4145
                     /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 accatgagcg tgaccggctt caccatcaca gacgagaagg tgcacctgta
      721 ccactccatc gagaaggaga agacagtgag gcacatcggc gatctgtgca gctcccacag
      781 cgtgaagaag atccaagtgg gcatctgcct gctgctggtg gagctgtgcg agcggttcac
      841 cttcttcgag gtggtgtgca acatgatccc tttctgtaca atcaagctgg gctaccacaa
      901 ctgccaggcc gccatcctga acctgtgctt tatcggcacc tccatcctga caccagtgtt
      961 cgtgaggtgg ctgaccgacg tgtacctggg ccgcaataag ctggtgtata tctgtctgtt
     1021 cctgcacttt ctgggaacag ccctgctgag cgtggtggca tttcccctgg aggatttcta
     1081 cctgggcacc tatcacgccg tgaacaatat ccctaagaca gagcagcaca ggctgtttta
     1141 cgtggccctg ctgaccatct gcctgggaat cggaggcgtg cgcgcaatcg tgtgccccct
     1201 gggagccttc ggcctgcagg agtacggaag ccagaagacc atgtccttct ttaactggtt
     1261 ttattggctg atgaacctga atgccacaat cgtgttcctg ggcatctctt acatccagca
     1321 cagccaggca tgggcactgg tgctgctgat cccattcatg tccatgctga tggccgtgat
     1381 caccctgcac atgatctact ataacctgat ctatcagtcc gagaagcggt gctctctgct
     1441 gaccggagtg ggcgtgctgg tgtctgccct gaagacatgc cacccccagt actgtcacct
     1501 gggcagagac gtgacctccc agctggatca cgccaaggag aagaatggcg gctgctattc
     1561 tgagctgcac gtggaggaca ccacattctt tctgaccctg ctgccactgt tcatctttca
     1621 gctgctgtac cggatgtgca tcatgcagat cccctctggc tactatctgc agacaatgaa
     1681 cagcaatctg aacctggatg gcttcctgct gcccatcgcc gtgatgaacg ccatctctag
     1741 cctgccactg ctgatcctgg ccccctttct ggagtatttc agcacctgcc tgtttccttc
     1801 caagcgggtg ggctctttcc tgagcacatg tatcatcgcc ggcaatctgt ttgccgccct
     1861 gtctgtgatg atcgccggct tctttgagat ccacagaaag cacttcccag cagtggagca
     1921 gcctctgagc ggcaaggtgc tgaccgtgtc ctctatgcct tgcttttacc tgatcctgca
     1981 gtatgtgctg ctgggcgtgg ccgagacact ggtgaaccct gccctgtccg tgatctctta
     2041 ccggtttgtg ccatctaatg tgagaggcac cagcatgaac ttcctgacac tgttcaatgg
     2101 ctttggctgt ttcaccggcg ccctgctggt gaagctggtg tatctgatct ccgacggcaa
     2161 ctggttccca aataccctga acaagggcaa tctggagagc ttctttttct ttctggcctc
     2221 cctgacactg ctgaatgtgc tgggcttttg cagcgtgtcc cagcggtact gtaatctgaa
     2281 ccacttcaac gcccagaata tcagaggcag caacctggag gagaccctgc tgctgcacga
     2341 gaagtccctg aagttttatg gctctatcca ggagttcagc tcctctatcg atctgtggga
     2401 gacagccctg gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     2461 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     2521 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     2581 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     2641 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     2701 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     2761 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     2821 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     2881 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     2941 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     3001 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     3061 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     3121 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     3181 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     3241 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     3301 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     3361 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     3421 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     3481 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     3541 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     3601 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3661 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3721 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3781 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3841 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3901 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3961 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4021 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4081 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4141 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     4201 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4261 cttttgctgg ccttttgctc acatgtt
//
