LOCUS       sequence-354278-        3444 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors. Contains a STOP codon at the end of 
            the codon-optimized ORF sequence. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3444)
  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 3444)
  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-SLC25A52_STOP
FEATURES             Location/Qualifiers
     source          1..3444
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1001..1100)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1117..1136)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1118..1136)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1141..1157)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1141..1157)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1259..1278)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1270..2079
                     /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(1337..1356)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2226..2814
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2715..2734
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2968..2985
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3144..3171)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3263..3349)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3398..3420
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3412..3429
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3413..3429
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacacat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccaccatgat cgacagcgag
      121 gcacacgaga agaggccacc tatcctgaca agctccaagc aggatatctc ccctcacatc
      181 accaacgtgg gcgagatgaa gcactacctg tgcggctgct gtgccgcctt caacaatgtg
      241 gccatcacat acccaatcca gaaggtgctg tttcggcagc agctgtatgg catcaagacc
      301 agagacgccg tgctgcagct gcggagagat ggcttccgga atctgtacag aggcatcctg
      361 ccacccctga tgcagaagac cacaaccctg gccctgatgt ttggcctgta tgaggacctg
      421 tcctgcctgc tgaggaagca cgtgcgcgca ccagagttcg caacacacgg agtggcagcc
      481 gtgctggcag gaacagccga ggccatcttt accccactgg agagggtgca gaccctgctg
      541 cagaaccaca agcaccacga taagttcaca aatacctatc aggcctttaa ggccctgaag
      601 tgtcacggca tcggcgagta ctatcggggc ctggtgccca tcctgttcag aaacggcctg
      661 tctaatgtgc tgttctttgg cctgaggggc cctatcaagg agcacctgcc aacagccaca
      721 acccacagcg cccacctggt gaacgacttt atcggaggag gactgctggg agcaatgctg
      781 ggcttcctgt gctttcccat caatgtggtg aagaccaggc tgcagtccca gatcggagga
      841 gagttccagt cttttcctaa ggtgttccag aagatctggc tggagaggga tcgcaagctg
      901 atcaacctgt ttaggggcgc ccacctgaat taccaccgct ctctgatcag ctggggcatc
      961 atcaacgcca cctatgagtt cctgctgaag tttatctaag acccagcttt cttgtacaaa
     1021 gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt
     1081 caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc gtattacatg
     1141 gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca
     1201 caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca
     1261 aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg
     1321 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
     1381 atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
     1441 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
     1501 cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact
     1561 gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
     1621 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt
     1681 ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt
     1741 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
     1801 aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc
     1861 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga
     1921 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
     1981 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
     2041 aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg
     2101 taacactggc agagcattac gctgacttga cgggacggcg caagctcatg accaaaatcc
     2161 cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2221 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2281 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2341 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     2401 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2461 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2521 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2581 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2641 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2701 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2761 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2821 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2881 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     2941 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     3001 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     3061 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg taccgctagc
     3121 caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttagttt
     3181 gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa
     3241 cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac
     3301 agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga tgcctggcag
     3361 ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg ttgtaaaacg
     3421 acggccagtc ttaagctcgg gccc
//
