LOCUS       sequence-259275-        4281 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 4281)
  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 4281)
  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_SLC15A4
FEATURES             Location/Qualifiers
     source          1..4281
                     /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(2406..2505)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2522..2541)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2523..2541)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2546..2562)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2546..2562)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2664..2683)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2675..3484
                     /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(2742..2761)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3631..4219
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4120..4139
                     /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 accatggagg gcagcggcgg cggcgccggc gagagggccc ccctgctggg
      721 cgcccggaga gccgccgcag cagcagcagc agcaggagcc tttgccggca ggcgcgccgc
      781 ctgcggagcc gtgctgctga cagagctgct ggagagggca gccttttatg gcattacatc
      841 caacctggta ctgttcctga atggcgcccc attttgttgg gagggagcac aggcaagcga
      901 ggccctgctg ctgttcatgg gcctgacata cctgggctcc ccctttggcg gctggctggc
      961 agacgcaagg ctgggaaggg caagggccat cctgctgagc ctggccctgt atctgctggg
     1021 aatgctggca tttccactgc tggcagcacc tgcaacaagg gccgccctgt gcggctctgc
     1081 cagactgctg aactgtaccg caccaggacc agatgcagca gcaaggtgct gtagcccagc
     1141 cacattcgcc ggactggtgc tggtgggact gggagtggca acagtgaagg ccaatatcac
     1201 cccctttggc gccgaccagg tgaaggatag aggccctgag gccacccgga gattctttaa
     1261 ctggttctac tggtctatca atctgggagc aatcctgagc ctgggaggaa tcgcctacat
     1321 ccagcagaac gtgtcctttg tgacaggcta tgcaatccca accgtgtgcg tgggactggc
     1381 cttcgtggtg ttcctgtgcg gccagtccgt gttcatcacc aagccccctg acggctctgc
     1441 cttcacagat atgtttaaga tcctgaccta tagctgctgt tcccagaaga ggtccggcga
     1501 gcgccagtct aatggcgagg gcatcggcgt gttccagcag agctccaagc agtccctgtt
     1561 tgactcttgc aagatgagcc acggcggccc cttcacagag gagaaggtgg aggatgtgaa
     1621 ggccctggtg aagatcgtgc cagtgttcct ggccctgatc ccctactgga ccgtgtattt
     1681 tcagatgcag accacatacg tgctgcagag cctgcacctg cggatccctg agatctccaa
     1741 catcaccaca acccctcaca cactgccagc cgcctggctg accatgtttg acgccgtgct
     1801 gatcctgctg ctgatccctc tgaaggacaa gctggtggat ccaatcctga ggcgccacgg
     1861 cctgctgcct tctagcctga agaggatcgc cgtgggcatg ttctttgtga tgtgctctgc
     1921 cttcgccgca ggaatcctgg agagcaagcg cctgaatctg gtgaaggaga agacaatcaa
     1981 ccagaccatc ggcaatgtgg tgtaccacgc cgccgatctg tccctgtggt ggcaggtgcc
     2041 tcagtatctg ctgatcggca tcagcgagat cttcgcctcc atcgccggcc tggagtttgc
     2101 ctattctgcc gccccaaagt ctatgcagag cgccatcatg ggcctgttct ttttcttttc
     2161 cggagtggga tcttttgtgg gaagcggact gctggccctg gtgagcatca aggccatcgg
     2221 ctggatgtcc tctcacacag acttcggcaa catcaatggc tgctacctga actactattt
     2281 ctttctgctg gcagcaatcc agggagccac cctgctgctg ttcctgatca tctccgtgaa
     2341 gtatgaccac cacagagatc accagcggtc cagagccaat ggcgtgccaa cctctcggag
     2401 agccgaccca gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     2461 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     2521 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     2581 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     2641 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     2701 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     2761 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     2821 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     2881 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     2941 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     3001 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     3061 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     3121 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     3181 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     3241 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     3301 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     3361 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     3421 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     3481 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     3541 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     3601 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3661 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3721 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     3781 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3841 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3901 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3961 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4021 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4081 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4141 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4201 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4261 ctggcctttt gctcacatgt t
//
