LOCUS       sequence-261797-        3471 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 3471)
  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 3471)
  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_SLC25A53
FEATURES             Location/Qualifiers
     source          1..3471
                     /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(1596..1695)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1712..1731)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1713..1731)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1736..1752)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1736..1752)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1854..1873)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1865..2674
                     /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(1932..1951)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2821..3409
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3310..3329
                     /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 accatgggcg agcagaacca cagcccaggc aaggagctgc agcacaggac
      721 cagagcagag gcacctggca agaagagctg gcactcccag gcatacgcac tgggagccgt
      781 gagcaatttc atgtccacct tcctgacatt tccaatctat aaggtggtgt ttcggcagca
      841 gatccacgcc atggccgtgt ccgaggccgt gcggcagctg tggcacgagg gacctcagta
      901 cttctataga ggcatctacc cccctctgct gtctaagacc ctgcagggca cactgctgtt
      961 cggcacctat gactccctgc tgtgctttct gtctccagtg ggaccacaca cactgggaca
     1021 cagatgggcc gccggcctga tgtctggagt ggtggaggca gtggccctga gcccatttga
     1081 gcgcgtgcag aacgtgctgc aggatggccg gaagcaggcc agattcccca gcaccttcag
     1141 catcctgaag gagttcaata gctacggact gtggggccgg ctgtccctgg gctactatag
     1201 aggcttttgg cctgtgctgg ccaggaactc cctgggctct gccctgtatt tctcctttaa
     1261 ggacccaatc caggatggac tggcagagca gggactgcct cactgggtgc cagccctggt
     1321 gtctggcagc gtgaatggca ccatcacatg tctggtgctg taccccctga tcgtgctggt
     1381 ggccaacatg cagagccaca tcggctggca gaatatgcct tctctgtggg ccagcgccca
     1441 ggacgtgtgg aacaccaggg gacgcaagct gctgctgatc tataggggag gatccctggt
     1501 catcctgcgc agctccgtga catggggcct gaccacagcc atccacgact tcctgcagag
     1561 gaagtcccac tctcgcaagg agctgaagac cgatgaccca gctttcttgt acaaagttgg
     1621 cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa
     1681 taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat
     1741 agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga
     1801 taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg
     1861 tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc
     1921 tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta
     1981 tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt
     2041 tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct
     2101 tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat
     2161 ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt
     2221 tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt
     2281 taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt
     2341 tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga
     2401 aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact
     2461 tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg
     2521 aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc
     2581 ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt
     2641 gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca
     2701 ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa
     2761 cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     2821 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     2881 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2941 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     3001 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3061 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3121 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3181 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     3241 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     3301 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3361 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3421 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt t
//
