LOCUS       sequence-262671-        4986 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 4986)
  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 4986)
  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_SLC9A2
FEATURES             Location/Qualifiers
     source          1..4986
                     /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(3111..3210)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3227..3246)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3228..3246)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3251..3267)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3251..3267)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3369..3388)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3380..4189
                     /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(3447..3466)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4336..4924
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4825..4844
                     /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 accatggagc ctctgggcaa ctggcgctcc ctgagggcac cactgccacc
      721 tatgttatta ctgctgctgc tgcaagtggc aggaccagtg ggcgccctgg ccgagacact
      781 gctgaatgca cctagggcaa tgggcaccag ctcctctcca ccaagcccag catccgtggt
      841 ggcaccagga accacactgt tcgaggagag ccggctgccc gtgtttacac tggattaccc
      901 tcacgtgcag atcccattcg agatcaccct gtggatcctg ctggcctctc tggccaagat
      961 cggctttcac ctgtatcaca agctgccaac aatcgtgccc gagagctgcc tgctgatcat
     1021 ggtgggcctg ctgctgggcg gcatcatctt cggcgtggat gagaagtccc ctccagccat
     1081 gaagaccgac gtgttctttc tgtacctgct gccacctatc gtgctggacg caggctattt
     1141 catgcccacc cgccctttct ttgagaacat cggcacaatc ttttggtacg ccgtggtggg
     1201 caccctgtgg aattctatcg gcatcggcgt gagcctgttc ggcatttgtc aaatagaggc
     1261 ctttggcctg agcgatatca ccctgctgca gaacctgctg ttcggctctc tgatcagcgc
     1321 cgtggaccct gtggccgtgc tggccgtgtt cgagaacatc cacgtgaatg agcagctgta
     1381 catcctggtg tttggcgaga gcctgctgaa cgatgccgtg accgtggtgc tgtataatct
     1441 gttcaagtcc ttttgtcaga tgaagaccat cgagacaatc gacgtgtttg ccggcatcgc
     1501 caacttcttt gtggtgggaa tcggaggcgt gctgatcgga atcttcctgg gctttatcgc
     1561 cgccttcacc acacgcttta cacacaatat cagagtgatc gagcccctgt tcgtgtttct
     1621 gtactcctat ctgtcttaca tcaccgccga gatgttccac ctgtccggca tcatggccat
     1681 caccgcctgc gccatgacaa tgaacaagta tgtggaggag aatgtgagcc agaagtccta
     1741 caccacaatc aagtacttca tgaagatgct gagctccgtg tctgagacac tgatcttcat
     1801 ctttatgggc gtgagcaccg tgggcaagaa ccacgagtgg aattgggcct tcgtgtgctt
     1861 tacactggcc ttctgtctga tgtggagggc cctgggcgtg ttcgtgctga cccaggtcat
     1921 caaccgcttt cggaccatcc cactgacatt caaggaccag tttatcatcg catacggagg
     1981 actgagggga gcaatctgtt tcgcactggt gtttctgctg ccagcagccg tgttccctcg
     2041 gaagaagctg tttatcaccg ccgccatcgt ggtcatcttc ttcaccgtgt tcatcctggg
     2101 catcacaatc agacccctgg tggagtttct ggatgtgaag aggtctaata agaagcagca
     2161 ggccgtgagc gaggagatct attgcagact gttcgatcac gtgaagaccg gaatcgagga
     2221 cgtgtgcgga cactggggcc acaacttttg gagggacaag ttcaagaagt ttgacgataa
     2281 gtacctgaga aagctgctga tcagggagaa tcagcctaag tctagcatcg tgtccctgta
     2341 taagaagctg gagatcaagc acgccatcga gatggccgag acaggcatga tcagcaccgt
     2401 gccaacattc gcctccctga acgattgtcg cgaggagaag atccggaagg tgacctcctc
     2461 tgagacagac gagatcagag agctgctgtc caggaacctg taccagatca gacagaggac
     2521 cctgagctat aatcgccact ccctgaccgc cgatacatct gagcggcagg ccaaggagat
     2581 cctgatccgg agaaggcact ctctgagaga gagcatcagg aaggacagct ccctgaacag
     2641 agagcacagg gcctccacct ctacaagcag atacctgagc ctgcctaaga atacaaagct
     2701 gccagagaag ctgcagaagc gccggaccat cagcatcgcc gatggcaact ctagcgactc
     2761 cgatgcagac gcaggaacca cagtgctgaa tctgcagcca agggccagaa ggttcctgcc
     2821 cgagcagttt tctaagaaga gcccccagtc ctataagatg gagtggaaga atgaggtgga
     2881 tgtggactcc ggcagagaca tgccctctac cccacccaca cctcactcca gggagaaggg
     2941 cacccagaca tctggcctgc tgcagcagcc tctgctgtcc aaggatcagt ccggctctga
     3001 gagagaggac tctctgaccg agggcatccc tccaaagcca cctccaaggc tggtgtggag
     3061 ggcatccgag ccaggatcta ggaaggcaag gttcggaagc gagaagcccg acccagcttt
     3121 cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt
     3181 cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc
     3241 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     3301 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     3361 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     3421 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     3481 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     3541 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     3601 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     3661 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     3721 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     3781 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     3841 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     3901 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     3961 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     4021 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     4081 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     4141 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     4201 taattggttg taacactggc agagcattac gctgacttga cgggacggcg caagctcatg
     4261 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     4321 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4381 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     4441 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     4501 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4561 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4621 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4681 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4741 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4801 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4861 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4921 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4981 catgtt
//
