LOCUS       sequence-260016-        3627 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 3627)
  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 3627)
  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_SLC25A39
FEATURES             Location/Qualifiers
     source          1..3627
                     /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(1752..1851)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1868..1887)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1869..1887)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1892..1908)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1892..1908)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2010..2029)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2021..2830
                     /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(2088..2107)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2977..3565
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3466..3485
                     /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 accatggcag accaggatcc agcaggaatc agccctctgc agcagatggt
      721 ggcatccgga acaggagcag tggtgacctc tctgttcatg acacctctgg acgtggtgaa
      781 ggtgaggctg cagagccagc gcccaagcat ggcctccgag ctgatgccca gctccagact
      841 gtggtccctg tcttacacca agctgccatc tagcctgcag tccaccggca agtgcctgct
      901 gtactgtaac ggcgtgctgg agccactgta tctgtgcccc aatggcgcca ggtgtgccac
      961 ctggttccag gaccccacac gctttaccgg cacaatggat gcttttgtga aaatcgtaag
     1021 acatgagggc accagaacac tgtggtctgg cctgcctgcc accctggtca tgacagtgcc
     1081 agccaccgcc atctacttca ccgcctatga ccagctgaag gcctttctgt gcggccgggc
     1141 cctgacaagc gatctgtacg caccaatggt ggcaggcgcc ctggcacggc tgggcaccgt
     1201 gacagtgatc tcccctctgg agctgatgag aaccaagctg caggcccagc acgtgtctta
     1261 tagggagctg ggagcatgcg tgcggaccgc agtggcccag ggcggctgga ggtccctgtg
     1321 gctgggatgg ggacctacag ccctgaggga cgtgccattc agcgccctgt actggtttaa
     1381 ctatgagctg gtgaagtcct ggctgaatgg cttccggcct aaggatcaga cctctgtggg
     1441 catgagcttt gtggcaggag gaatcagcgg aaccgtggca gccgtgctga cactgccatt
     1501 cgacgtggtg aagacacaga ggcaggtggc cctgggagca atggaggccg tgagagtgaa
     1561 ccccctgcac gtggattcta cctggctgct gctgcggaga atcagggccg agagcggcac
     1621 aaagggcctg ttcgccggct ttctgccccg catcatcaag gccgcccctt cctgcgccat
     1681 catgatctct acctatgagt ttggcaagtc cttctttcag aggctgaatc aggatcgcct
     1741 gctgggcggc gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     1801 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     1861 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     1921 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     1981 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     2041 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     2101 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     2161 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     2221 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     2281 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     2341 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     2401 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     2461 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     2521 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     2581 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     2641 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     2701 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     2761 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     2821 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     2881 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     2941 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3001 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3061 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3121 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3181 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3241 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3301 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3361 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3421 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3481 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3541 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3601 cttttgctgg ccttttgctc acatgtt
//
