LOCUS       sequence-261199-        4338 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 4338)
  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 4338)
  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_SLC5A10
FEATURES             Location/Qualifiers
     source          1..4338
                     /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(2463..2562)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2579..2598)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2580..2598)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2603..2619)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2603..2619)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2721..2740)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2732..3541
                     /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(2799..2818)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3688..4276
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4177..4196
                     /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 accatggccg ccaactccac atctgacctg cacacccctg gcacacagct
      721 gtccgtggcc gatatcatcg tgatcaccgt gtacttcgcc ctgaatgtgg cagtgggaat
      781 ctggagctcc tgcagggcct ctagaaacac agtgaatggc tattttctgg caggaaggga
      841 catgacctgg tggccaatcg gagcatccct gttcgcatct agcgagggct ctggcctgtt
      901 tatcggactg gcaggaagcg gagcagcagg aggcctggca gtggcaggat tcgagtggaa
      961 cgcaacatac gtgctgctgg ccctggcctg ggtgtttgtg ccaatctata tctcctctga
     1021 gatcgtgacc ctgcccgagt acatccagaa gagatatggc ggccagcgga tcagaatgta
     1081 cctgtccgtg ctgtctctgc tgctgagcgt gttcaccaag atcagcctgg acctgtacgc
     1141 aggcgccctg ttcgtgcaca tctgtctggg ctggaacttc tacctgtcta ccatcctgac
     1201 actgggcatc accgccctgt acacaatcgc aggaggactg gcagccgtga tctataccga
     1261 cgccctgcag acactgatca tggtggtggg cgccgtgatc ctgaccatca aggcctttga
     1321 tcagatcgga ggatacggac agctggaggc agcatatgca caggcaatcc catccaggac
     1381 catcgcaaac accacatgcc acctgcctag gacagatgcc atgcacatgt tcagggaccc
     1441 acacaccggc gatctgcctt ggaccggcat gacatttggc ctgaccatca tggccacatg
     1501 gtactggtgt accgaccagg tcatcgtgca gaggagcctg tccgcccgcg atctgaatca
     1561 cgcaaaggca ggaagcatcc tggcatccta tctgaagatg ctgccaatgg gcctgatcat
     1621 catgccagga atgatcagcc gggccctgtt ccctgacgat gtgggctgcg tggtgccaag
     1681 cgagtgcctg agagcatgtg gagcagaagt gggctgttcc aacatcgcct accccaagct
     1741 ggtcatggag ctgatgccta tcggcctgcg cggcctgatg atcgccgtga tgctggccgc
     1801 cctgatgagc tccctgacct ccatcttcaa ttctagctcc accctgttta caatggacat
     1861 ctggcggaga ctgaggccac gctctggaga gagggagctg ctgctggtgg gccggctggt
     1921 catcgtggcc ctgatcggcg tgagcgtggc atggatcccc gtgctgcagg attctaacag
     1981 cggccagctg tttatctaca tgcagagcgt gacctctagc ctggcaccac ctgtgacagc
     2041 cgtgttcgtg ctgggcgtgt tttggaggag ggcaaacgag cagggagcat tctggggact
     2101 gatcgcagga ctggtggtgg gagcaacaag gctggtgctg gagtttctga atccagcacc
     2161 accatgcgga gagcctgata ccaggccagc cgtgctggga tccatccact atctgcactt
     2221 cgcagtggcc ctgtttgcac tgagcggagc agtggtggtg gcaggatccc tgctgacccc
     2281 tccaccccag tctgtgcaga tcgagaatct gacctggtgg acactggcac aggacgtgcc
     2341 tctgggaacc aaggcaggcg atggacagac accacagaag cacgcattct gggcacgggt
     2401 gtgcggcttt aacgccatcc tgctgatgtg cgtgaatatc ttcttttacg cctatttcgc
     2461 cgacccagct ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg
     2521 caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc
     2581 ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca
     2641 aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc
     2701 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag
     2761 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     2821 tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt
     2881 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     2941 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
     3001 tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga
     3061 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     3121 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
     3181 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     3241 ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc
     3301 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
     3361 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     3421 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     3481 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     3541 atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg
     3601 cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga
     3661 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3721 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3781 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3841 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3901 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3961 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4021 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4081 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4141 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4201 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4261 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4321 gccttttgct cacatgtt
//
