LOCUS       sequence-345426-        5241 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors. Contains a STOP codon at the end of 
            the codon-optimized ORF sequence. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5241)
  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 5241)
  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-SLC9A5_STOP
FEATURES             Location/Qualifiers
     source          1..5241
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     RBS             2004..2012
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     protein_bind    complement(2798..2897)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2914..2933)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2915..2933)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2938..2954)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2938..2954)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3056..3075)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3067..3876
                     /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(3134..3153)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4023..4611
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4512..4531
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4765..4782
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4941..4968)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5060..5146)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5195..5217
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5209..5226
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5210..5226
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacacat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccaccatgct gagagccgcc
      121 ctgtctctgc tggccctgcc actggcagga gcagcagagg agcctaccca gaagccagag
      181 agcccaggag agccacctcc aggactggag ctgttcagat ggcagtggca cgaggtggag
      241 gcaccatacc tggtggccct gtggatcctg gtggcaagcc tggccaagat cgtgtttcac
      301 ctgtctagga aggtgaccag cctggtgcct gagtcctgcc tgctgatcct gctgggactg
      361 gtgctgggag gaatcgtgct ggccgtggcc aagaaggccg agtatcagct ggagccaggc
      421 acattctttc tgttcctgct gccccctatc gtgctggact ccggctactt tatgccctct
      481 cgcctgttct ttgataacct gggcgccatc ctgacctacg cagtggttgg gactttgtgg
      541 aatgccttca ccacaggagc cgccctgtgg ggactgcagc aggcaggcct ggtggcccct
      601 agggtgcagg caggactgct ggacttcctg ctgtttggca gcctgatctc cgccgtggat
      661 ccagtggccg tgctggccgt gtttgaggag gtgcacgtga acgagaccct gttcatcatc
      721 gtgtttggcg agagcctgct gaatgacgcc gtgacagtgg tgctgtacaa ggtgtgcaac
      781 agcttcgtgg agatgggctc cgccaatgtg caggccaccg attatctgaa gggcgtggcc
      841 tctctgtttg tggtgagcct gggaggagca gcagtgggac tggtgttcgc ctttctgctg
      901 gccctgacca caagattcac aaagagagtg aggatcatcg agcctctgct ggtgtttctg
      961 ctggcctacg ccgcctatct gaccgccgag atggcctctc tgagcgccat cctggccgtg
     1021 acaatgtgcg gcctgggctg taagaagtac gtggaggcca acatctctca caagagcagg
     1081 accacagtga agtataccat gaagacactg gcctcctgtg ccgagaccgt gatcttcatg
     1141 ctgctgggca tctccgccgt ggacagctcc aagtgggcat gggattctgg cctggtgctg
     1201 ggcaccctga tcttcatcct gttctttcgc gccctgggag tggtgctgca gacatgggtg
     1261 ctgaatcagt ttcggctggt gccactggac aagatcgatc aggtggtcat gagctacgga
     1321 ggactgaggg gagcagtggc cttcgccctg gtcatcctgc tggaccggac caaggtgccc
     1381 gccaaggatt attttgtggc caccacaatc gtggtggtgt tctttaccgt gatcgtgcag
     1441 ggcctgacaa tcaagcccct ggtgaagtgg ctgaaggtga agcggtccga gcaccacaag
     1501 cctaccctga accaggagct gcacgagcac acattcgacc acatcctggc agcagtggag
     1561 gatgtggtgg gacaccacgg ctaccactat tggcgcgacc ggtgggagca gtttgataag
     1621 aagtacctgt cccagctgct gatgcggaga tctgcctata gaatcaggga ccagatctgg
     1681 gacgtgtact atcgcctgaa tatccgggac gccatcagct tcgtggatca gggaggacac
     1741 gtgctgtcta gcaccggact gacactgcct tccatgccat ctagaaacag cgtggccgag
     1801 acctccgtga caaatctgct gagggagtcc ggatctggag catgcctgga cctgcaggtc
     1861 atcgataccg tgcgctctgg aagggacagg gaggatgccg tgatgcacca cctgctgtgc
     1921 ggaggactgt acaagccaag gcgccggtat aaggccagct gttccagaca cttcatctct
     1981 gaggacgccc aggagaggca ggataaggag gtgtttcagc agaacatgaa gagaaggctg
     2041 gagtccttca agtctaccaa gcacaatatc tgttttacaa agtccaagcc tagaccaagg
     2101 aagaccggcc gccggaagaa ggacggagtg gcaaacgcag aggcaacaaa tggcaagcac
     2161 aggggactgg gattccagga caccgcagcc gtgatcctga cagtggagag cgaggaggag
     2221 gaggaggagt ccgattcctc tgagaccgag aaggaggacg atgagggcat catctttgtg
     2281 gcaagggcca caagcgaggt gctgcaggag ggcaaggtgt ctggaagcct ggaggtgtgc
     2341 ccatctccta ggatcatccc acccagccca acctgtgccg agaaggagct gccatggaag
     2401 agcggacagg gcgacctggc cgtgtacgtg agctccgaga ccacaaagat cgtgcctgtg
     2461 gatatgcaga caggctggaa ccagtccatc tctagcctgg agagcctggc ctcccctcca
     2521 tgcaatcagg cccctatcct gacctgtctg ccccctcacc caagaggcac agaggagccc
     2581 caggtgcctc tgcacctgcc aagcgacccc aggtcctctt tcgcctttcc accatccctg
     2641 gcaaaggcag gccgctcccg gtctgagagc tccgccgacc tgcctcagca gcaggagctg
     2701 cagccactga tgggccacaa ggatcacacc cacctgtctc caggcaccgc cacaagccac
     2761 tggtgtatcc agttcaacag aggctccagg ctgtaagacc cagctttctt gtacaaagtt
     2821 ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa
     2881 aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta ttacatggtc
     2941 atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa
     3001 gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg
     3061 ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat
     3121 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
     3181 tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc
     3241 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct
     3301 cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg
     3361 atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
     3421 gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct
     3481 tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg
     3541 gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa
     3601 gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca
     3661 cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc
     3721 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
     3781 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
     3841 ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa
     3901 cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc aaaatccctt
     3961 aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4021 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4081 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4141 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4201 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4261 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4321 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4381 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4441 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4501 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4561 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4621 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     4681 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     4741 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     4801 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     4861 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atacgcgtac cgctagccag
     4921 gaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttagtttgat
     4981 gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt
     5041 tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga
     5101 taaaacgaaa ggcccagtct tccgactgag cctttcgttt tatttgatgc ctggcagttc
     5161 cctactctcg cgttaacgct agcatggatg ttttcccagt cacgacgttg taaaacgacg
     5221 gccagtctta agctcgggcc c
//
