LOCUS       sequence-261794-        3636 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 3636)
  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 3636)
  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_SLC25A3
FEATURES             Location/Qualifiers
     source          1..3636
                     /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(1761..1860)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1877..1896)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1878..1896)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1901..1917)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1901..1917)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2019..2038)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2030..2839
                     /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(2097..2116)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2986..3574
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3475..3494
                     /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 accatgttca gctccgtggc acacctggca agggcaaacc cttttaatac
      721 cccacacctg cagctggtgc acgacggact gggcgatctg cggtctagct cccctggacc
      781 aacaggacag ccacggagac ccagaaacct ggcagcagca gcagtggagg agcagtactc
      841 ctgtgactat ggctctggcc ggttctttat cctgtgcggc ctgggcggca tcatctcctg
      901 tggaaccaca cacaccgccc tggtgccact ggacctggtg aagtgcagaa tgcaggtgga
      961 tcctcagaag tacaagggca tcttcaatgg cttttccgtg accctgaagg aggatggcgt
     1021 gaggggactg gcaaagggat gggcacctac attcctgggc tatagcatgc agggcctgtg
     1081 caagttcggc ttttacgagg tgtttaaggt gctgtatagc aacatgctgg gcgaggagaa
     1141 tacctacctg tggaggacat ccctgtatct ggcagcatcc gcctctgccg agttctttgc
     1201 agacatcgcc ctggcaccaa tggaggcagc aaaggtgcgg atccagaccc agcctggcta
     1261 cgccaacaca ctgagagatg ccgccccaaa gatgtacaag gaggagggcc tgaaggcctt
     1321 ctataagggc gtggcccccc tgtggatgag gcagatccct tacaccatga tgaagttcgc
     1381 ctgctttgag cgcacagtgg aggccctgta taagttcgtg gtgcccaagc ctcggagcga
     1441 gtgttccaag ccagagcagc tggtggtgac cttcgtggcc ggctacatcg ccggcgtgtt
     1501 ttgcgcaatc gtgtcccacc cagcagactc tgtggtgagc gtgctgaata aggagaaggg
     1561 ctctagcgcc tctctggtgc tgaagaggct gggcttcaag ggcgtgtgga agggcctgtt
     1621 tgcccgcatc atcatgatcg gcaccctgac agccctgcag tggttcatct acgattctgt
     1681 gaaggtgtac ttccggctgc caaggccacc tccaccagag atgcccgaga gcctgaagaa
     1741 gaagctgggc ctgacacagg acccagcttt cttgtacaaa gttggcatta taagaaagca
     1801 ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg
     1861 ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca
     1921 gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     1981 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     2041 tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     2101 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa
     2161 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     2221 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     2281 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc
     2341 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     2401 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     2461 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     2521 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt
     2581 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     2641 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     2701 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     2761 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     2821 gctcgatgag tttttctaat cagaattggt taattggttg taacactggc agagcattac
     2881 gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc
     2941 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3001 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3061 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3121 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3181 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3241 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3301 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3361 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3421 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     3481 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     3541 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     3601 gttcctggcc ttttgctggc cttttgctca catgtt
//
