LOCUS       sequence-261006-        3441 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 3441)
  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 3441)
  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_SLC25A51
FEATURES             Location/Qualifiers
     source          1..3441
                     /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(1566..1665)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1682..1701)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1683..1701)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1706..1722)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1706..1722)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1824..1843)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1835..2644
                     /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(1902..1921)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2791..3379
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3280..3299
                     /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 accatgatgg actctgaggc ccacgagaag aggcccccta tcctgacaag
      721 ctccaagcag gatatcagcc ctcacatcac caacgtgggc gagatgaagc actacctgtg
      781 cggctgctgt gccgccttca acaatgtggc catcacattt ccaatccaga aggtgctgtt
      841 caggcagcag ctgtacggca tcaagaccag ggacgcaatc ctgcagctga ggagagatgg
      901 ctttcggaat ctgtatagag gcatcctgcc acccctgatg cagaagacca caaccctggc
      961 cctgatgttc ggcctgtacg aggacctgag ctgtctgctg cacaagcacg tgtccgcccc
     1021 agagtttgcc acatctggag tggcagccgt gctggcagga acaaccgagg ccatcttcac
     1081 accactggag agggtgcaga ccctgctgca ggaccacaag caccacgata agtttacaaa
     1141 cacctatcag gccttcaagg ccctgaagtg ccacggcatc ggcgagtact ataggggcct
     1201 ggtgcccatc ctgtttcgca acggcctgtc caatgtgctg ttctttggcc tgcggggccc
     1261 tatcaaggag cacctgccaa cagccacaac ccactctgcc cacctggtga acgactttat
     1321 ctgtggagga ctgctgggag caatgctggg cttcctgttc tttcccatca atgtggtgaa
     1381 gaccagaatc cagagccaga tcggcggcga gttccagtcc tttcctaagg tgtttcagaa
     1441 gatctggctg gagagggatc gcaagctgat caacctgttc cggggcgccc acctgaatta
     1501 ccacagatct ctgatcagct ggggcatcat caatgccacc tatgagttcc tgctgaaagt
     1561 gatcgaccca gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     1621 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     1681 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     1741 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     1801 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     1861 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     1921 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     1981 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     2041 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     2101 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     2161 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     2221 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     2281 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     2341 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     2401 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     2461 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     2521 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     2581 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     2641 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     2701 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     2761 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2821 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2881 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     2941 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3001 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3061 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3121 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3181 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3241 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3301 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3361 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3421 ctggcctttt gctcacatgt t
//
