LOCUS       sequence-260461-        5238 bp    DNA     circular SYN 14-APR-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 5238)
  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 5238)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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
FEATURES             Location/Qualifiers
     source          1..5238
                     /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"
     RBS             2572..2580
                     /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(3363..3462)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3479..3498)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3480..3498)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3503..3519)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3503..3519)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3621..3640)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3632..4441
                     /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(3699..3718)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4588..5176
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5077..5096
                     /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 accatgctga gagccgccct gtctctgctg gccctgccac tggcaggagc
      721 agcagaggag cctacccaga agccagagag cccaggagag ccacctccag gactggagct
      781 gttcagatgg cagtggcacg aggtggaggc accatacctg gtggccctgt ggatcctggt
      841 ggcaagcctg gccaagatcg tgtttcacct gtctaggaag gtgaccagcc tggtgcctga
      901 gtcctgcctg ctgatcctgc tgggactggt gctgggagga atcgtgctgg ccgtggccaa
      961 gaaggccgag tatcagctgg agccaggcac attctttctg ttcctgctgc cccctatcgt
     1021 gctggactcc ggctacttta tgccctctcg cctgttcttt gataacctgg gcgccatcct
     1081 gacctacgca gtggttggga ctttgtggaa tgccttcacc acaggagccg ccctgtgggg
     1141 actgcagcag gcaggcctgg tggcccctag ggtgcaggca ggactgctgg acttcctgct
     1201 gtttggcagc ctgatctccg ccgtggatcc agtggccgtg ctggccgtgt ttgaggaggt
     1261 gcacgtgaac gagaccctgt tcatcatcgt gtttggcgag agcctgctga atgacgccgt
     1321 gacagtggtg ctgtacaagg tgtgcaacag cttcgtggag atgggctccg ccaatgtgca
     1381 ggccaccgat tatctgaagg gcgtggcctc tctgtttgtg gtgagcctgg gaggagcagc
     1441 agtgggactg gtgttcgcct ttctgctggc cctgaccaca agattcacaa agagagtgag
     1501 gatcatcgag cctctgctgg tgtttctgct ggcctacgcc gcctatctga ccgccgagat
     1561 ggcctctctg agcgccatcc tggccgtgac aatgtgcggc ctgggctgta agaagtacgt
     1621 ggaggccaac atctctcaca agagcaggac cacagtgaag tataccatga agacactggc
     1681 ctcctgtgcc gagaccgtga tcttcatgct gctgggcatc tccgccgtgg acagctccaa
     1741 gtgggcatgg gattctggcc tggtgctggg caccctgatc ttcatcctgt tctttcgcgc
     1801 cctgggagtg gtgctgcaga catgggtgct gaatcagttt cggctggtgc cactggacaa
     1861 gatcgatcag gtggtcatga gctacggagg actgagggga gcagtggcct tcgccctggt
     1921 catcctgctg gaccggacca aggtgcccgc caaggattat tttgtggcca ccacaatcgt
     1981 ggtggtgttc tttaccgtga tcgtgcaggg cctgacaatc aagcccctgg tgaagtggct
     2041 gaaggtgaag cggtccgagc accacaagcc taccctgaac caggagctgc acgagcacac
     2101 attcgaccac atcctggcag cagtggagga tgtggtggga caccacggct accactattg
     2161 gcgcgaccgg tgggagcagt ttgataagaa gtacctgtcc cagctgctga tgcggagatc
     2221 tgcctataga atcagggacc agatctggga cgtgtactat cgcctgaata tccgggacgc
     2281 catcagcttc gtggatcagg gaggacacgt gctgtctagc accggactga cactgccttc
     2341 catgccatct agaaacagcg tggccgagac ctccgtgaca aatctgctga gggagtccgg
     2401 atctggagca tgcctggacc tgcaggtcat cgataccgtg cgctctggaa gggacaggga
     2461 ggatgccgtg atgcaccacc tgctgtgcgg aggactgtac aagccaaggc gccggtataa
     2521 ggccagctgt tccagacact tcatctctga ggacgcccag gagaggcagg ataaggaggt
     2581 gtttcagcag aacatgaaga gaaggctgga gtccttcaag tctaccaagc acaatatctg
     2641 ttttacaaag tccaagccta gaccaaggaa gaccggccgc cggaagaagg acggagtggc
     2701 aaacgcagag gcaacaaatg gcaagcacag gggactggga ttccaggaca ccgcagccgt
     2761 gatcctgaca gtggagagcg aggaggagga ggaggagtcc gattcctctg agaccgagaa
     2821 ggaggacgat gagggcatca tctttgtggc aagggccaca agcgaggtgc tgcaggaggg
     2881 caaggtgtct ggaagcctgg aggtgtgccc atctcctagg atcatcccac ccagcccaac
     2941 ctgtgccgag aaggagctgc catggaagag cggacagggc gacctggccg tgtacgtgag
     3001 ctccgagacc acaaagatcg tgcctgtgga tatgcagaca ggctggaacc agtccatctc
     3061 tagcctggag agcctggcct cccctccatg caatcaggcc cctatcctga cctgtctgcc
     3121 ccctcaccca agaggcacag aggagcccca ggtgcctctg cacctgccaa gcgaccccag
     3181 gtcctctttc gcctttccac catccctggc aaaggcaggc cgctcccggt ctgagagctc
     3241 cgccgacctg cctcagcagc aggagctgca gccactgatg ggccacaagg atcacaccca
     3301 cctgtctcca ggcaccgcca caagccactg gtgtatccag ttcaacagag gctccaggct
     3361 ggacccagct ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg
     3421 caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc
     3481 ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca
     3541 aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc
     3601 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag
     3661 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     3721 tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt
     3781 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     3841 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
     3901 tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga
     3961 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     4021 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
     4081 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     4141 ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc
     4201 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
     4261 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     4321 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     4381 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     4441 atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg
     4501 cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga
     4561 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4621 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4681 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     4741 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4801 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4861 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4921 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4981 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     5041 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     5101 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     5161 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     5221 gccttttgct cacatgtt
//
