LOCUS       sequence-262886-        4593 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 4593)
  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 4593)
  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_SLC5A9
FEATURES             Location/Qualifiers
     source          1..4593
                     /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"
     regulatory      689..698
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2718..2817)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2834..2853)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2835..2853)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2858..2874)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2858..2874)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2976..2995)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2987..3796
                     /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(3054..3073)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3943..4531
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4432..4451
                     /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 accatgagca aggagctggc cgccatggga ccaggagcat ccggcgacgg
      721 cgtgcggacc gagacagccc cccacatcgc cctggactcc agagtgggac tgcacgccta
      781 cgatatctct gtggtggtca tctacttcgt gtttgtgatc gccgtgggca tctggagctc
      841 catcagggca agcaggggaa ccatcggagg atacttcctg gccggcagat ccatgtcttg
      901 gtggcccatc ggcgcctctc tgatgtctag caacgtggga agcggactgt ttatcggact
      961 ggcaggcaca ggagcagcag gaggactggc agtgggagga ttcgagtgga atgcaacctg
     1021 gctgctgctg gccctgggat gggtgtttgt gcccgtgtac atcgcagcag gagtggtgac
     1081 aatgcctcag tatctgaaga agcggttcgg cggccagaga atccaggtgt acatgagcgt
     1141 gctgtccctg atcctgtata tctttacaaa gatctctacc gatatcttca gcggcgccct
     1201 gtttatccag atggccctgg gctggaacct gtatctgagc accggcatcc tgctggtggt
     1261 gacagccgtg tacaccatcg ccggcggcct gatggccgtg atctatacag acgccctgca
     1321 gaccgtgatc atggtgggag gcgccctggt gctgatgttc ctgggctttc aggatgtggg
     1381 atggtaccca ggactggagc agcggtatag acaggccatc cccaacgtga cagtgcctaa
     1441 taccacatgc cacctgccaa ggccagacgc attccacatc ctgagggacc ccgtgagcgg
     1501 cgatatccct tggccaggcc tgatctttgg cctgacagtg ctggccacct ggtgctggtg
     1561 taccgatcag gtcatcgtgc agcggagcct gtccgccaag tctctgagcc acgcaaaggg
     1621 aggatccgtg ctgggaggct acctgaagat cctgcccatg ttctttatcg tgatgcctgg
     1681 catgatctct agagccctgt tcccagacga agtgggatgc gtggacccag acgtgtgcca
     1741 gaggatctgt ggagcaaggg tgggatgtag caacatcgca tatcctaagc tggtcatggc
     1801 actgatgcca gtgggactga ggggactgat gatcgccgtg atcatggccg ccctgatgtc
     1861 ctctctgaca agcatcttca atagctcctc taccctgttt acaatcgacg tgtggcagag
     1921 gttccggaga aagtccaccg agcaggagct gatggtggtg ggacgcgtgt tcgtggtgtt
     1981 tctggtggtc atctctatcc tgtggatccc tatcatccag agctccaaca gcggccagct
     2041 gtttgattac atccaggccg tgacatccta tctggccccc cctatcaccg ccctgttcct
     2101 gctggccatc ttttgcaaga gggtgaccga gccaggagca ttctggggac tggtgtttgg
     2161 actgggagtg ggactgctga gaatgatcct ggagttctcc taccccgccc ctgcatgtgg
     2221 agaggtggac aggcgccccg ccgtgctgaa ggatttccac tacctgtatt ttgccatcct
     2281 gctgtgcggc ctgacagcca tcgtgatcgt gatcgtgtct ctgtgcacca caccaatccc
     2341 cgaggagcag ctgacccggc tgacatggtg gaccagaaac tgccctctgt ccgagctgga
     2401 gaaggaggcc cacgagtcca ccccagagat ctctgagagg ccagcaggag agtgtccagc
     2461 aggaggagga gcagccgaga attctagcct gggacaggag cagcctgagg caccatcccg
     2521 ctcttggggc aagctgctgt ggagctggtt ctgcggactg tccggaacac ctgagcaggc
     2581 cctgtctcca gcagagaagg ccgccctgga gcagaagctg accagcatcg aggaggagcc
     2641 actgtggagg cacgtgtgca acatcaatgc cgtgctgctg ctggccatca atatcttcct
     2701 gtggggctac tttgccgacc cagctttctt gtacaaagtt ggcattataa gaaagcattg
     2761 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     2821 tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     2881 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     2941 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     3001 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     3061 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     3121 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     3181 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     3241 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
     3301 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     3361 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     3421 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     3481 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     3541 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     3601 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     3661 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     3721 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     3781 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     3841 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     3901 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3961 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     4021 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     4081 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     4141 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     4201 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     4261 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     4321 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     4381 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     4441 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     4501 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     4561 cctggccttt tgctggcctt ttgctcacat gtt
//
