LOCUS       sequence-260426-        3957 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 3957)
  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 3957)
  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_SLC25A25
FEATURES             Location/Qualifiers
     source          1..3957
                     /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(2082..2181)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2198..2217)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2199..2217)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2222..2238)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2222..2238)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2340..2359)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2351..3160
                     /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(2418..2437)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3307..3895
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3796..3815
                     /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 accatgctgt gcctgtgcct gtacgtgcca gtgatcggcg aggcccagac
      721 agagttccag tattttgagt ccaagggcct gcccgccgag ctgaagagca tcttcaagct
      781 gtccgtgttt atcccttccc aggagttttc tacctaccgg cagtggaagc agaagatcgt
      841 gcaggccggc gacaaggatc tggacggcca gctggacttc gaggagtttg tgcactatct
      901 gcaggatcac gagaagaagc tgagactggt gttcaagtcc ctggataaga agaacgacgg
      961 ccggatcgat gcccaggaga tcatgcagtc tctgagagac ctgggcgtga agatcagcga
     1021 gcagcaggcc gagaagatcc tgaagtctat ggataagaac ggcaccatga caatcgactg
     1081 gaatgagtgg cgggattacc acctgctgca ccccgtggag aacatccctg agatcatcct
     1141 gtattggaag cacagcacaa tctttgacgt gggcgagaat ctgaccgtgc ctgatgagtt
     1201 cacagtggag gagaggcaga ccggaatgtg gtggaggcac ctggtggcag gaggaggagc
     1261 aggagccgtg agccggacct gcacagcccc actggacaga ctgaaggtgc tgatgcaggt
     1321 gcacgccagc aggtccaaca atatgggcat cgtgggcggc tttacacaga tgatcaggga
     1381 gggaggagca cggtctctgt ggagaggcaa cggcatcaat gtgctgaaga tcgcccccga
     1441 gagcgccatc aagttcatgg cctacgagca gatcaagagg ctggtgggct ccgaccagga
     1501 gaccctgagg atccacgagc gcctggtggc aggatctctg gcaggagcaa tcgcacagag
     1561 ctccatctac cctatggagg tgctgaagac acggatggcc ctgagaaaga ccggccagta
     1621 ttccggcatg ctggattgtg caaggagaat cctggcaagg gagggagtgg ccgcctttta
     1681 caagggctat gtgccaaata tgctgggcat catcccctac gccggcatcg acctggccgt
     1741 gtatgagaca ctgaagaacg cctggctgca gcactacgcc gtgaattctg ccgatcctgg
     1801 cgtgttcgtg ctgctggcat gcggaaccat gtctagcaca tgtggccagc tggccagcta
     1861 tccactggcc ctggtgagga ccaggatgca ggcacaggca tccatcgagg gagcacccga
     1921 ggtgacaatg tcctctctgt tcaagcacat cctgaggacc gagggagcct ttggcctgta
     1981 caggggactg gcccctaact tcatgaaagt gatcccagcc gtgtctatca gctacgtggt
     2041 gtatgagaat ctgaagatca ccctgggcgt gcagtccaga gacccagctt tcttgtacaa
     2101 agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag
     2161 tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat
     2221 ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc
     2281 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
     2341 aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat
     2401 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     2461 aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     2521 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     2581 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     2641 tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     2701 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     2761 tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
     2821 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     2881 gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt
     2941 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     3001 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     3061 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     3121 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     3181 gtaacactgg cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc
     3241 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3301 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3361 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3421 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3481 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3541 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3601 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3661 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3721 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3781 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3841 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3901 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
