LOCUS       sequence-390327-        3486 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 3486)
  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 3486)
  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-SLC25A36_STOP
FEATURES             Location/Qualifiers
     source          1..3486
                     /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"
     regulatory      478..487
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1043..1142)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1159..1178)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1160..1178)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1183..1199)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1183..1199)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1301..1320)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1312..2121
                     /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(1379..1398)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2268..2856
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2757..2776
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3010..3027
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3186..3213)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3305..3391)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3440..3462
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3454..3471
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3455..3471
                     /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 ccaccatgtc ccagagggac
      121 acactggtgc acctgttcgc aggaggatgc ggaggaaccg tgggagccat cctgacatgt
      181 ccactggagg tggtgaagac cagactgcag agctcctctg tgacactgta catctccgag
      241 gtgcagctga acaccatggc cggcgcctcc gtgaataggg tggtgtctcc aggacctctg
      301 cactgcctga aagtgatcct ggagaaggag ggaccacgct ctctgttccg gggactgggc
      361 ccaaacctgg tgggagtggc accatccagg gcaatctact tcgccgccta ttctaactgt
      421 aaggagaagc tgaatgacgt gttcgaccct gattctaccc aggtgcacat gatcagcgcc
      481 gccatggccg gctttacagc catcaccgcc acaaacccaa tctggctgat caagaccagg
      541 ctgcagctgg acgcaaggaa taggggagag aggagaatgg gagccttcga gtgcgtgcgc
      601 aaggtgtatc agacagatgg cctgaagggc ttttaccggg gcatgagcgc ctcctatgcc
      661 ggcatcagcg agaccgtgat ccacttcgtg atctacgagt ccatcaagca gaagctgctg
      721 gagtataaga ccgcctctac aatggagaat gacgaggagt ctgtgaagga ggccagcgat
      781 tttgtgggaa tgatgctggc agcagccacc agcaagacat gtgccaccac aatcgcatac
      841 ccacacgagg tggtgcggac cagactgagg gaggagggca caaagtatcg gagcttcttt
      901 cagaccctgt ccctgctggt gcaggaggag ggctacggca gcctgtatag aggcctgacc
      961 acacacctgg tgaggcagat ccctaacacc gccatcatga tggccacata cgagctggtg
     1021 gtgtatctgc tgaatggcta agacccagct ttcttgtaca aagttggcat tataagaaag
     1081 cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt
     1141 tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc tgtttcctgg
     1201 cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca
     1261 tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat
     1321 attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga tttatatggg
     1381 tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg
     1441 aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt
     1501 acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag
     1561 cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca
     1621 gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca
     1681 gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc
     1741 gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat
     1801 tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt
     1861 ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt
     1921 tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga
     1981 taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa
     2041 cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg
     2101 atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg gcagagcatt
     2161 acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt gagttacgcg
     2221 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     2281 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     2341 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     2401 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     2461 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     2521 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     2581 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     2641 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     2701 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     2761 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     2821 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     2881 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     2941 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     3001 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     3061 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     3121 gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta gccaggaaga gtttgtagaa
     3181 acgcaaaaag gccatccgtc aggatggcct tctgcttagt ttgatgcctg gcagtttatg
     3241 gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac aacgttcaaa tccgctcccg
     3301 gcggatttgt cctactcagg agagcgttca ccgacaaaca acagataaaa cgaaaggccc
     3361 agtcttccga ctgagccttt cgttttattt gatgcctggc agttccctac tctcgcgtta
     3421 acgctagcat ggatgttttc ccagtcacga cgttgtaaaa cgacggccag tcttaagctc
     3481 gggccc
//
