LOCUS       sequence-262667-        4668 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 4668)
  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 4668)
  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_SLC6A9
FEATURES             Location/Qualifiers
     source          1..4668
                     /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(2793..2892)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2909..2928)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2910..2928)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2933..2949)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2933..2949)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3051..3070)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3062..3871
                     /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(3129..3148)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4018..4606
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4507..4526
                     /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 accatgagcg gaggcgacac cagggcagca atcgcacggc ccagaatggc
      721 cgccgcccac ggccctgtgg ccccaagctc ccccgagcag gtgacactgc tgccagtgca
      781 gagatccttc tttctgccac ctttctctgg agcaacccca tccacatctc tggcagagtc
      841 cgtgctgaag gtctggcacg gcgcctacaa ctctggcctg ctgcctcagc tgatggccca
      901 gcacagcctg gcaatggcac agaacggagc agtgccatcc gaggcaacca agcgggatca
      961 gaatctgaag agaggcaact ggggcaatca gatcgagttt gtgctgacat ccgtgggata
     1021 cgcagtggga ctgggaaacg tgtggaggtt cccatacctg tgctatcgca atggcggcgg
     1081 cgcctttatg ttcccctact tcatcatgct gatcttctgt ggcatccctc tgttctttat
     1141 ggagctgagc tttggccagt tcgcatccca gggatgcctg ggcgtgtgga ggatctctcc
     1201 tatgtttaag ggcgtgggct acggcatgat ggtggtgagc acctatatcg gcatctacta
     1261 taacgtggtc atctgtatcg ccttctacta tttcttttct agcatgacac acgtgctgcc
     1321 ctgggcctac tgcaacaatc cttggaatac ccacgactgt gccggcgtgc tggatgcctc
     1381 taacctgaca aatggaagca gaccagccgc cctgccatcc aacctgtctc acctgctgaa
     1441 tcactccctg cagcggacca gcccatccga ggagtactgg agactgtatg tgctgaagct
     1501 gtctgacgat atcggcaatt ttggagaggt gcggctgcca ctgctgggat gcctgggcgt
     1561 gagctggctg gtggtgttcc tgtgcctgat cagaggcgtg aagtcctctg gcaaggtggt
     1621 gtactttacc gccacattcc cttatgtggt gctgaccatc ctgtttgtga ggggagtgac
     1681 actggaggga gccttcgacg gcatcatgta ctatctgacc ccacagtggg acaagatcct
     1741 ggaggcaaaa gtgtggggcg atgcagcatc ccagatcttt tactctctgg gatgcgcatg
     1801 gggaggactg atcacaatgg ccagctacaa caagttccac aacaattgct atagggatag
     1861 cgtgatcatc tccatcacca attgtgccac aagcgtgtac gccggctttg tgatcttctc
     1921 catcctgggc tttatggcaa accacctggg agtggacgtg tccagggtgg cagatcacgg
     1981 accaggactg gccttcgtgg catatccaga ggccctgacc ctgctgccta tctctccact
     2041 gtggagcctg ctgttctttt tcatgctgat cctgctgggc ctgggcacac agttctgtct
     2101 gctggagacc ctggtgacag ccatcgtgga cgaagtgggc aatgagtgga tcctgcagaa
     2161 gaagacctac gtgacactgg gagtggcagt ggcaggcttt ctgctgggaa tcccactgac
     2221 ctctcaggca ggcatctact ggctgctgct gatggacaac tatgccgcct ctttcagcct
     2281 ggtggtcatc tcctgcatca tgtgcgtggc catcatgtac atctatggcc acaggaatta
     2341 ttttcaggat atccagatga tgctgggctt cccaccccct ctgtttttcc agatctgctg
     2401 gcgctttgtg agccccgcca tcatcttttt catcctggtg ttcaccgtga tccagtacca
     2461 gcctatcaca tataaccact accagtatcc aggctgggcc gtggccatcg gctttctgat
     2521 ggccctgagc tccgtgctgt gcatccctct gtacgccatg ttcaggctgt gccgcaccga
     2581 cggcgataca ctgctgcaga ggctgaagaa tgccaccaag cctagcaggg actggggacc
     2641 agccctgctg gagcaccgga ccggcagata tgcaccaaca atcgcaccat ccccagagga
     2701 cggattcgag gtgcagccac tgcaccctga taaggcccag atccctatcg tgggctctaa
     2761 cggctctagc aggctgcagg atagccgcat cgacccagct ttcttgtaca aagttggcat
     2821 tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa
     2881 aatcattatt tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc
     2941 tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     3001 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
     3061 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
     3121 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     3181 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     3241 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     3301 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     3361 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     3421 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     3481 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     3541 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     3601 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     3661 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     3721 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     3781 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     3841 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg
     3901 gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt
     3961 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4021 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4081 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4141 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4201 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4261 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4321 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4381 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4441 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4501 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4561 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4621 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtt
//
