LOCUS       sequence-261163-        4455 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 4455)
  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 4455)
  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_SLC5A6
FEATURES             Location/Qualifiers
     source          1..4455
                     /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(2580..2679)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2696..2715)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2697..2715)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2720..2736)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2720..2736)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2838..2857)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2849..3658
                     /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(2916..2935)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3805..4393
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4294..4313
                     /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 accatgtccg tgggcgtgtc cacatctgcc ccactgtccc ctacctctgg
      721 cacaagcgtg ggcatgtcca ccttttctat catggactac gtggtgttcg tgctgctgct
      781 ggtgctgtct ctggcaatcg gactgtatca cgcatgcagg ggatggggcc ggcacaccgt
      841 gggagagctg ctgatggccg atcggaagat gggctgtctg cctgtggccc tgtctctgct
      901 ggccacattc cagagcgccg tggcaatcct gggagtgcca tccgagatct acagattcgg
      961 cacccagtat tggtttctgg gctgctgtta cttcctgggc ctgctgatcc ctgcccacat
     1021 cttcatccca gtgttttata ggctgcacct gacaagcgcc tacgagtatc tggagctgcg
     1081 gttcaacaag accgtgaggg tgtgcggcac cgtgacattc atctttcaga tggtcatcta
     1141 catgggagtg gtgctgtatg caccatccct ggccctgaat gcagtgaccg gcttcgacct
     1201 gtggctgtct gtgctggccc tgggcatcgt gtgcaccgtg tacacagccc tgggaggcct
     1261 gaaggccgtg atctggacag acgtgttcca gaccctggtc atgttcctgg gccagctggc
     1321 cgtgatcatc gtgggctccg ccaaagtggg aggactggga cgcgtgtggg cagtggccag
     1381 ccagcacggc cggatctccg gctttgagct ggaccctgat ccattcgtgc gccacacctt
     1441 ttggacactg gccttcggcg gcgtgtttat gatgctgagc ctgtacggcg tgaaccaggc
     1501 acaggtgcag cggtacctga gctccaggac cgagaaggca gccgtgctgt cttgctacgc
     1561 cgtgttcccc tttcagcagg tgagcctgtg cgtgggctgt ctgatcggcc tggtcatgtt
     1621 cgcctactat caggagtacc ctatgtccat ccagcaggca caggcagcac cagaccagtt
     1681 cgtgctgtat tttgtgatgg atctgctgaa gggactgcca ggactgccag gactgttcat
     1741 cgcatgcctg tttagcggct ccctgtctac catctctagc gcctttaaca gcctggccac
     1801 cgtgacaatg gaggacctga tccggccttg gttcccagag ttttccgagg caagggcaat
     1861 catgctgtct aggggactgg ccttcggcta cggactgctg tgcctgggaa tggcctatat
     1921 ctcctctcag atgggccctg tgctgcaggc cgccatcagc atctttggaa tggtgggagg
     1981 accactgctg ggactgttct gcctgggcat gttctttcct tgtgccaatc cacctggagc
     2041 agtggtggga ctgctggcag gcctggtcat ggccttttgg atcggcatcg gcagcatcgt
     2101 gacatccatg ggcagctcca tgccacccag cccatccaac ggctctagct tcagcctgcc
     2161 caccaatctg acagtggcca ccgtgaccac actgatgccc ctgaccacat tttccaagcc
     2221 taccggcctg cagagattct actctctgag ctacctgtgg tattctgccc acaacagcac
     2281 cacagtgatc gtggtgggcc tgatcgtgag cctgctgaca ggccggatga gaggcaggtc
     2341 cctgaatcca gccaccatct accccgtgct gcctaagctg ctgtccctgc tgcccctgtc
     2401 ttgccagaag agactgcact gtaggtctta tggccaggac cacctggata caggcctgtt
     2461 cccagagaag cccagaaatg gcgtgctggg cgacagcagg gataaggagg ccatggccct
     2521 ggatggcacc gcctatcagg gctcctctag cacatgtatc ctgcaggaga ccagcctgga
     2581 cccagctttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa
     2641 cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg atatccccta
     2701 tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa
     2761 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
     2821 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc
     2881 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     2941 cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt
     3001 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     3061 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     3121 tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata
     3181 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     3241 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
     3301 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     3361 gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt
     3421 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     3481 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     3541 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     3601 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     3661 agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc
     3721 aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc
     3781 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3841 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3901 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     3961 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4021 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4081 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     4141 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     4201 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4261 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     4321 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     4381 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     4441 ttttgctcac atgtt
//
