LOCUS       sequence-262890-        5052 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 5052)
  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 5052)
  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_SLC9A3
FEATURES             Location/Qualifiers
     source          1..5052
                     /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(3177..3276)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3293..3312)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3294..3312)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3317..3333)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3317..3333)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3435..3454)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3446..4255
                     /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(3513..3532)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4402..4990
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4891..4910
                     /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 accatgtggg gcctgggcgc cagaggccca gacaggggcc tgctgctggc
      721 cctggccctg ggaggactgg caagggcagg aggcgtggag gtggagcccg gcggcgccca
      781 cggcgagtct ggcggctttc aggtggtgac cttcgagtgg gcccacgtgc aggatcctta
      841 tgtgatcgcc ctgtggatcc tggtggcaag cctggccaag atcggcttcc acctgtctca
      901 caaggtgacc agcgtcgtgc cagagtccgc cctgctgatc gtgctgggac tggtgctggg
      961 aggaatcgtg tgggccgccg accacatcgc cagctttacc ctgacaccca ccgtgttctt
     1021 tttctacctg ctgcccccta tcgtgctgga tgccggctat ttcatgccta accggctgtt
     1081 tttcggcaat ctgggcacaa tcctgctgta tgcagtggtg ggaaccgtgt ggaacgcagc
     1141 aaccacagga ctgagcctgt acggcgtgtt tctgtccgga ctgatgggcg acctgcagat
     1201 cggactgctg gattttctgc tgttcggaag cctgatggca gcagtggacc ctgtggccgt
     1261 gctggccgtg ttcgaggagg tgcacgtgaa cgaggtgctg tttatcatcg tgttcggcga
     1321 gtccctgctg aacgatgccg tgacagtggt gctgtacaac gtgttcgaga gttttgtcgc
     1381 acttggtggg gacaatgtga ccggcgtgga ttgcgtgaag ggcatcgtga gcttcttcgt
     1441 ggtgagcctg ggaggcacac tggtgggcgt ggtgtttgcc ttcctgctgt ccctggtgac
     1501 acgctttacc aagcacgtgc ggatcatcga gcccggcttc gtgttcatca tctcctacct
     1561 gtcttatctg acatctgaga tgctgtctct gagcgccatc ctggccatca ccttctgtgg
     1621 catctgctgt cagaagtacg tgaaggccaa catcagcgag cagtccgcca ccacagtgag
     1681 atatacaatg aagatgctgg ccagctccgc cgagaccatc atctttatgt tcctgggcat
     1741 cagcgccgtg aaccccttca tctggacatg gaataccgcc tttgtgctgc tgacactggt
     1801 gttcatctcc gtgtacaggg caatcggagt ggtgctgcag acctggctgc tgaacagata
     1861 taggatggtg cagctggagc ccatcgacca ggtggtgctg tcctacggag gactgagggg
     1921 agcagtggcc ttcgccctgg tggtgctgct ggacggcgat aaggtgaagg agaagaatct
     1981 gtttgtgagc accacaatca tcgtggtgtt tttcacagtg atcttccagg gcctgaccat
     2041 caagccactg gtgcagtggc tgaaggtgaa gagatccgag cacagggagc cacggctgaa
     2101 cgagaagctg cacggcaggg cctttgacca catcctgtcc gccatcgagg atatctctgg
     2161 ccagatcggc cacaattacc tgagggacaa gtggtcccac tttgatcgca agttcctgtc
     2221 tcgggtgctg atgcggagaa gcgcccagaa gtccagggac cggatcctga acgtgttcca
     2281 cgagctgaat ctgaaggatg ccatcagcta tgtggcagag ggagagagga ggggatccct
     2341 ggcctttatc agatccccct ctaccgacaa cgtggtgaat gtggatttca cacctcgctc
     2401 tagcaccgtg gaggccagcg tgtcctacct gctgcgggag aacgtgagcg ccgtgtgcct
     2461 ggacatgcag tctctggagc agcggagaag gagcatcagg gacgccgagg atatggtgac
     2521 acaccacacc ctgcagcagt acctgtataa gcctagacag gagtacaagc acctgtattc
     2581 caggcacgag ctgacaccaa ccgaggacga gaagcaggat agagagatct ttcacaggac
     2641 aatgagaaag aggctggagt ctttcaagag caccaagctg ggcctgaacc agaataagaa
     2701 ggccgccaag ctgtataaga gggagagggc acagaagagg aggaactcct ctatccctaa
     2761 tggcaagctg cctatggagt ctccagccca gaacttcaca atcaaggaga aggacctgga
     2821 gctgagcgat accgaggagc cacccaatta cgacgaggag atgagcggcg gcatcgagtt
     2881 tctggcctcc gtgacaaagg acaccgcctc tgatagccca gccggcatcg ataatcccgt
     2941 gttctctcct gacgaggccc tggatcgcag cctgctggca aggctgcctc catggctgtc
     3001 cccaggagag accgtggtgc catctcagag agccaggaca cagatcccat acagccccgg
     3061 caccttttgt agactgatgc ccttcaggct gagctccaag tccgtggact ctttcctgca
     3121 ggccgatgga cctgaggaga ggccacctgc cgccctgcca gagtccaccc acatggaccc
     3181 agctttcttg tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga
     3241 acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag
     3301 tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct
     3361 ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta
     3421 cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg
     3481 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
     3541 gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct
     3601 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     3661 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
     3721 atggttactc accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc
     3781 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
     3841 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
     3901 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     3961 tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt
     4021 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     4081 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     4141 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     4201 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga
     4261 attggttaat tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag
     4321 ctcatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt
     4381 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     4441 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4501 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     4561 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4621 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4681 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4741 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4801 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4861 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4921 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4981 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     5041 tgctcacatg tt
//
