LOCUS       sequence-479344-        3825 bp    DNA     circular SYN 26-JUL-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 3825)
  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 3825)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 26, 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-SLC35A5_STOP
FEATURES             Location/Qualifiers
     source          1..3825
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     112..131
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     365..382
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(541..568)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(660..746)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     795..817
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     809..826
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     810..826
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    842..941
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2223..2322)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2339..2358)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2340..2358)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2363..2379)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2363..2379)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2481..2500)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2492..3301
                     /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(2559..2578)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(3448..3825,1..211)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
       61 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
      121 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
      181 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
      241 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
      301 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
      361 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
      421 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
      481 cagtgagcgc aacgcaatta atacgcgtac cgctagccag gaagagtttg tagaaacgca
      541 aaaaggccat ccgtcaggat ggccttctgc ttagtttgat gcctggcagt ttatggcggg
      601 cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga
      661 tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct
      721 tccgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg cgttaacgct
      781 agcatggatg ttttcccagt cacgacgttg taaaacgacg gccagtctta agctcgggcc
      841 ccaaataatg attttatttt gactgatagt gacctgttcg ttgcaacaca ttgatgagca
      901 atgctttttt ataatgccaa ctttgtacaa aaaagcaggc tccaccatgg agaagcagtg
      961 ctgtagccac cctgtgatct gctccctgtc taccatgtac acattcctgc tgggcgccat
     1021 ctttatcgcc ctgagctcct ctcggatcct gctggtcaag tatagcgcca acgaggagaa
     1081 taagtacgac tatctgccaa ccacagtgaa cgtgtgctcc gagctggtga agctggtgtt
     1141 ctgcgtgctg gtgtcctttt gcgtgatcaa gaaggatcac cagtctagga atctgaagta
     1201 cgcctcttgg aaggagttca gcgactttat gaagtggtct atccccgcct tcctgtattt
     1261 tctggataac ctgatcgtgt tctacgtgct gagctatctg cagcctgcca tggccgtgat
     1321 cttcagcaat ttttccatca tcaccacagc cctgctgttc aggatcgtgc tgaagcggag
     1381 actgaactgg atccagtggg ccagcctgct gaccctgttt ctgtccatcg tggccctgac
     1441 agccggcacc aagacactgc agcacaacct ggcaggaagg ggattccacc acgacgcctt
     1501 ctttagccct tccaactctt gcctgctgtt taggagcgag tgcccacgca aggataattg
     1561 taccgccaag gagtggacat tccccgaggc caagtggaac accacagccc gggtgttttc
     1621 ccacatcaga ctgggcatgg gccacgtgct gatcatcgtg cagtgtttca tcagctccat
     1681 ggccaacatc tacaatgaga agatcctgaa ggagggcaac cagctgaccg agtccatctt
     1741 catccagaat tctaagctgt atttctttgg catcctgttt aatggcctga cactgggcct
     1801 gcagcggagc aacagagacc agatcaagaa ttgcggcttc ttttacggcc actctgcctt
     1861 cagcgtggcc ctgatcttcg tgaccgcctt tcagggcctg tccgtggcct tcatcctgaa
     1921 gtttctggac aacatgtttc acgtgctgat ggcccaggtg accacagtga tcatcaccac
     1981 agtgtctgtg ctggtgttcg attttcggcc aagcctggag ttctttctgg aggcccccag
     2041 cgtgctgctg tccatcttca tctacaatgc ctccaagccc caggtgcctg agtatgcccc
     2101 aagacaggag aggatccgcg acctgtctgg caacctgtgg gagaggtcta gcggcgatgg
     2161 agaggagctg gagcgcctga ccaagccaaa gtccgatgag tctgacgagg atacatttta
     2221 agacccagct ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg
     2281 caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc
     2341 ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca
     2401 aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc
     2461 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag
     2521 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     2581 tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt
     2641 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     2701 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
     2761 tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga
     2821 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     2881 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
     2941 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     3001 ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc
     3061 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
     3121 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     3181 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     3241 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     3301 atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg
     3361 cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga
     3421 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3481 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3541 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3601 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3661 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3721 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3781 cacacagccc agcttggagc gaacgaccta caccgaactg agata
//
