LOCUS       sequence-261072-        4557 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 4557)
  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 4557)
  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_SLC9A6
FEATURES             Location/Qualifiers
     source          1..4557
                     /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(2682..2781)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2798..2817)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2799..2817)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2822..2838)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2822..2838)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2940..2959)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2951..3760
                     /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(3018..3037)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3907..4495
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4396..4415
                     /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 accatggctc ggagaggctg gaggcgcgcc cctctgcgga gaggagtggg
      721 aagctcccct agggcaaggc gcctgatgag accactgtgg ctgctgctgg cagtgggcgt
      781 gttcgattgg gcaggagcat ctgacggagg aggaggagag gcccgggcca tggatgagga
      841 gatcgtgagc gagaagcagg ccgaggagtc ccacagacag gactctgcca atctgctgat
      901 cttcatcctg ctgctgacac tgaccatcct gaccatctgg ctgtttaagc accggagagc
      961 caggttcctg cacgagacag gcctggccat gatctacgga ctgctggtgg gactggtgct
     1021 gcgctatggc atccacgtgc catccgatgt gaacaatgtg accctgtctt gcgaggtgca
     1081 gtctagcccc accacactgc tggtgacatt cgaccctgag gtgttcttta atatcctgct
     1141 gccccctatc atcttttacg ccggctattc cctgaagagg cgccacttct ttcggaacct
     1201 gggctctatc ctggcctacg cctttctggg caccgccatc agctgcttcg tgatcggctc
     1261 catcatgtat ggctgcgtga ccctgatgaa ggtgaccggc cagctggccg gcgatttcta
     1321 ctttaccgac tgtctgctgt tcggcgccat cgtgtccgcc acagatccag tgaccgtgct
     1381 ggccatcttc cacgagctgc aggtggacgt ggagctgtat gccctgctgt ttggcgagag
     1441 cgtgctgaat gacgccgtgg ccatcgtgct gtcctctagc atcgtggcat accagccagc
     1501 aggcgataac tctcacacat ttgacgtgac cgccatgttc aagagcatcg gcatcttcct
     1561 gggcatcttt tctggcagct tcgccatggg agcagcaaca ggagtggtga ccgccctggt
     1621 gacaaagttt accaagctga gggagttcca gctgctggag acaggcctgt tctttctgat
     1681 gtcctggtct acctttctgc tggcagaggc atggggcttc acaggagtgg tggccgtgct
     1741 gttttgcggc atcacacagg cccactacac ctataacaat ctgagcacag agtcccagca
     1801 ccgcaccaag cagctgtttg agctgctgaa cttcctggcc gagaatttca tcttttccta
     1861 tatgggcctg acactgttca cctttcagaa ccacgtgttt aatcctacct tcgtggtggg
     1921 cgcctttgtg gccatcttcc tgggcagggc cgccaacatc tacccactga gcctgctgct
     1981 gaatctgggc cggagatcta agatcggcag caattttcag cacatgatga tgttcgcagg
     2041 actgagggga gcaatggcct tcgccctggc catcagggat acagccacct acgcccgcca
     2101 gatgatgttc tccaccacac tgctgatcgt gttctttacc gtgtgggtgt ttggaggagg
     2161 aaccacagca atgctgagct gcctgcacat ccgggtgggc gtggattccg accaggagca
     2221 cctgggagtg ccagagaacg agaggaggac cacaaaggca gagtccgcct ggctgtttag
     2281 aatgtggtac aacttcgacc acaattatct gaagccactg ctgacccact ctggaccacc
     2341 actgaccaca accctgccag catgctgtgg acctatcgca agatgtctga ccagccctca
     2401 ggcctatgag aaccaggagc agctgaagga cgatgactcc gatctgatcc tgaatgatgg
     2461 cgacatctct ctgacctacg gcgacagcac agtgaacacc gagccagcca catcctctgc
     2521 cccacggaga ttcatgggca acagctccga ggatgcactg gacagggagc tggcctttgg
     2581 cgatcacgag ctggtcatca ggggaaccag gctggtgctg ccaatggatg acagcgagcc
     2641 tccactgaac ctgctggaca atacaagaca cggccccgcc gacccagctt tcttgtacaa
     2701 agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag
     2761 tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat
     2821 ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc
     2881 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
     2941 aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat
     3001 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     3061 aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     3121 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     3181 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     3241 tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     3301 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     3361 tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
     3421 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     3481 gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt
     3541 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     3601 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     3661 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     3721 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     3781 gtaacactgg cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc
     3841 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3901 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3961 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4021 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4081 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4141 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4201 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4261 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4321 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4381 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4441 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4501 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
