LOCUS       sequence-260962-        4191 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 4191)
  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 4191)
  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_SLC22A25
FEATURES             Location/Qualifiers
     source          1..4191
                     /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(2316..2415)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2432..2451)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2433..2451)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2456..2472)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2456..2472)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2574..2593)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2585..3394
                     /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(2652..2671)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3541..4129
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4030..4049
                     /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 accatggcct ttcaggacct gctggatcaa gtgggaggac tgggccggtt
      721 ccagatcctg cagatggtgt ttctgatcat gttcaacgtg atcgtgtacc accagaccca
      781 gctggagaat ttcgccgcct ttatcctgga ccacagatgc tgggtgcaca tcctggacaa
      841 cgataccatc ccagacaatg atccaggcac actgagccag gacgcactgc tgaggatctc
      901 tatcccattt gatagcaacc tgcgccccga gaagtgtcgg agattcgtgc accctcagtg
      961 gaagctgatc cacctgaacg gcacctttcc aaatacatcc gagcctgaca cagagccatg
     1021 cgtggacggc tgggtgtacg atcagagctc cttcccctct accatcgtga caaagtggga
     1081 cctggtgtgc gagtcccagc ctctgaactc tgtggccaag ttcctgttta tggccggcat
     1141 gatggtgggc ggcaatctgt acggccacct gagcgatagg tttggccgca agttcgtgct
     1201 gaggtggtcc tatctgcagc tggcaatcgt gggaacctgc gcagccttcg cccctacaat
     1261 cctggtgtat tgttctctgc gctttctggc aggagcagcc accttcagca tcatcgtgaa
     1321 cacagtgctg ctgatcgtgg agtggatcac ccaccagttt tgcgccatgg cactgaccct
     1381 gaccctgtgc gcagcaagca tcggacacat cacactgggc tccctggcct tcgtgatccg
     1441 ggaccagtgc atcctgcagc tggtcatgtc cgccccttgt ttcgtgttct tcctgttcag
     1501 caggtggctg gcagagtccg ccaggtggct gatcatcaac aataagccag aggagggcct
     1561 gaaggagctg aggaaggcag cccacagaaa cggcatgaag aatgccgagg atatcctgac
     1621 catggaggtg ctgaagtcta caatgaagca ggagctggag gccgcccaga agaagcacag
     1681 cctgtgcgag ctgctgcgga tcccaaatat ctgcaagaga atctgtttcc tgtcttttgt
     1741 gagattcgcc agcaccatcc ccttctgggg cctgacactg cacctgcagc acctgggcaa
     1801 caacgtgttc ctgctgcaga ccctgttcgg cgccgtgaca ctgctggcaa actgcgtggc
     1861 accatgggcc ctgaatcaca tgagcaggcg cctgtcccag atgctgctga tgtttctgct
     1921 ggccacctgt ctgctggcca tcatcttcgt gcctcaggag atgcagaccc tgagggtggt
     1981 gctggccaca ctgggagtgg gagcagcatc cctgggaatc acctgttcta cagcccagga
     2041 gaacgagctg atcccaagca tcatccgggg cagagccacc ggcatcacag gcaactttgc
     2101 aaatatcgga ggcgccctgg catccctgat gatgatcctg tccatctact ctcgccccct
     2161 gccttggatc atctatggcg tgttcgccat cctgagcgga ctggtggtgc tgctgctgcc
     2221 cgagacccgg aatcagcctc tgctggactc catccaggat gtggagaacg agggcgtgaa
     2281 ttctctggcc gccccccaga gatctagcgt gctggaccca gctttcttgt acaaagttgg
     2341 cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa
     2401 taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat
     2461 agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga
     2521 taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg
     2581 tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc
     2641 tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta
     2701 tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt
     2761 tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct
     2821 tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat
     2881 ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt
     2941 tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt
     3001 taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt
     3061 tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga
     3121 aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact
     3181 tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg
     3241 aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc
     3301 ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt
     3361 gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca
     3421 ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa
     3481 cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3541 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     3601 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3661 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     3721 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3781 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3841 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3901 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     3961 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4021 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4081 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     4141 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt t
//
