LOCUS       sequence-334676-        3456 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 3456)
  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 3456)
  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-SLC25A20_STOP
FEATURES             Location/Qualifiers
     source          1..3456
                     /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(1013..1112)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1129..1148)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1130..1148)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1153..1169)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1153..1169)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1271..1290)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1282..2091
                     /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(1349..1368)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2238..2826
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2727..2746
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2980..2997
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3156..3183)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3275..3361)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3410..3432
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3424..3441
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3425..3441
                     /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 ccaccatggc cgaccagcca
      121 aagcccatca gccctctgaa gaacctgctg gcaggaggat tcggaggcgt gtgcctggtg
      181 tttgtgggcc accctctgga caccgtgaag gtgagactgc agacacagcc accttccctg
      241 ccaggacagc caccaatgta ctctggcacc ttcgattgct ttcgcaagac actgttccgg
      301 gagggcatca ccggactgta taggggaatg gcagcaccta tcatcggcgt gacaccaatg
      361 ttcgccgtgt gcttcttcgg ctttggcctg ggcaagaagc tgcagcagaa gcaccccgag
      421 gatgtgctga gctaccctca gctgttcgca gcaggaatgc tgtccggcgt gtttaccaca
      481 ggcatcatga ccccaggcga gagaatcaag tgcctgctgc agatccaggc aagctccgga
      541 gagtccaagt ataccggcac actggactgt gccaagaagc tgtaccagga gttcggcatc
      601 cgcggcatct ataagggcac cgtgctgaca ctgatgcggg atgtgcccgc ctctggcatg
      661 tacttcatga catatgagtg gctgaagaat atctttaccc cagagggcaa gagagtgtct
      721 gagctgagcg ccccaaggat cctggtggca ggaggcatcg ccggcatctt taactgggcc
      781 gtggccatcc ctccagacgt gctgaagtct cgcttccaga cagccccccc tggcaagtac
      841 cccaatggct ttcgggacgt gctgagagag ctgatcaggg atgagggcgt gaccagcctg
      901 tataagggct tcaacgccgt gatgatccgg gcctttcccg ccaatgccgc ctgtttcctg
      961 ggctttgagg tggccatgaa gttcctgaac tgggccaccc ctaatctgta agacccagct
     1021 ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag
     1081 gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc ctatagtgag
     1141 tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga
     1201 tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata
     1261 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta
     1321 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
     1381 tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa
     1441 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
     1501 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
     1561 ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat
     1621 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
     1681 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
     1741 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
     1801 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact
     1861 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
     1921 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     1981 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     2041 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg
     2101 gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca
     2161 tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa
     2221 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2281 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2341 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     2401 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     2461 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2521 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     2581 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     2641 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     2701 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     2761 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     2821 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     2881 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     2941 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     3001 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     3061 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg
     3121 cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct
     3181 tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc
     3241 gttgcttcac aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca
     3301 ccgacaaaca acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt
     3361 gatgcctggc agttccctac tctcgcgtta acgctagcat ggatgttttc ccagtcacga
     3421 cgttgtaaaa cgacggccag tcttaagctc gggccc
//
