LOCUS       sequence-479329-        4596 bp    DNA     circular SYN 06-AUG-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 4596)
  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 4596)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 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-SLC5A9_STOP
FEATURES             Location/Qualifiers
     source          1..4596
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(14..31)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(185..773)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(265..284)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(920..1729)
                     /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     1643..1662
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1721..1740
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1842..1858
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1842..1858
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1863..1882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1863..1881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1899..1998
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      4021..4030
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(4051..4150)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4166..4183)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4166..4182)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4175..4197)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      4246..4332
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4424..4451
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
       61 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
      121 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
      181 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
      241 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
      301 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
      361 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
      421 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
      481 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
      541 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
      601 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
      661 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
      721 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
      781 ctttgatctt ttctacgggg tctgacgctc agtggaacga cgcgtaactc acgttaaggg
      841 attttggtca tgagcttgcg ccgtcccgtc aagtcagcgt aatgctctgc cagtgttaca
      901 accaattaac caattctgat tagaaaaact catcgagcat caaatgaaac tgcaatttat
      961 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
     1021 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
     1081 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
     1141 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc
     1201 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
     1261 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
     1321 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
     1381 tttcacctga atcaggatat tcttctaata cctggaatgc tgtttttccg gggatcgcag
     1441 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
     1501 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
     1561 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaag cgatagattg
     1621 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
     1681 tgttggaatt taatcgcggc ctcgacgttt cccgttgaat atggctcata acaccccttg
     1741 tattactgtt tatgtaagca gacagtttta ttgttcatga tgatatattt ttatcttgtg
     1801 caatgtaaca tcagagattt tgagacacgg gccagagctg ccaggaaaca gctatgacca
     1861 tgtaatacga ctcactatag gggatatcag ctggatggca aataatgatt ttattttgac
     1921 tgatagtgac ctgttcgttg caacaaattg ataagcaatg ctttcttata atgccaactt
     1981 tgtacaagaa agctgggtct taggcaaagt agccccacag gaagatattg atggccagca
     2041 gcagcacggc attgatgttg cacacgtgcc tccacagtgg ctcctcctcg atgctggtca
     2101 gcttctgctc cagggcggcc ttctctgctg gagacagggc ctgctcaggt gttccggaca
     2161 gtccgcagaa ccagctccac agcagcttgc cccaagagcg ggatggtgcc tcaggctgct
     2221 cctgtcccag gctagaattc tcggctgctc ctcctcctgc tggacactct cctgctggcc
     2281 tctcagagat ctctggggtg gactcgtggg cctccttctc cagctcggac agagggcagt
     2341 ttctggtcca ccatgtcagc cgggtcagct gctcctcggg gattggtgtg gtgcacagag
     2401 acacgatcac gatcacgatg gctgtcaggc cgcacagcag gatggcaaaa tacaggtagt
     2461 ggaaatcctt cagcacggcg gggcgcctgt ccacctctcc acatgcaggg gcggggtagg
     2521 agaactccag gatcattctc agcagtccca ctcccagtcc aaacaccagt ccccagaatg
     2581 ctcctggctc ggtcaccctc ttgcaaaaga tggccagcag gaacagggcg gtgatagggg
     2641 gggccagata ggatgtcacg gcctggatgt aatcaaacag ctggccgctg ttggagctct
     2701 ggatgatagg gatccacagg atagagatga ccaccagaaa caccacgaac acgcgtccca
     2761 ccaccatcag ctcctgctcg gtggactttc tccggaacct ctgccacacg tcgattgtaa
     2821 acagggtaga ggagctattg aagatgcttg tcagagagga catcagggcg gccatgatca
     2881 cggcgatcat cagtcccctc agtcccactg gcatcagtgc catgaccagc ttaggatatg
     2941 cgatgttgct acatcccacc cttgctccac agatcctctg gcacacgtct gggtccacgc
     3001 atcccacttc gtctgggaac agggctctag agatcatgcc aggcatcacg ataaagaaca
     3061 tgggcaggat cttcaggtag cctcccagca cggatcctcc ctttgcgtgg ctcagagact
     3121 tggcggacag gctccgctgc acgatgacct gatcggtaca ccagcaccag gtggccagca
     3181 ctgtcaggcc aaagatcagg cctggccaag ggatatcgcc gctcacgggg tccctcagga
     3241 tgtggaatgc gtctggcctt ggcaggtggc atgtggtatt aggcactgtc acgttgggga
     3301 tggcctgtct ataccgctgc tccagtcctg ggtaccatcc cacatcctga aagcccagga
     3361 acatcagcac cagggcgcct cccaccatga tcacggtctg cagggcgtct gtatagatca
     3421 cggccatcag gccgccggcg atggtgtaca cggctgtcac caccagcagg atgccggtgc
     3481 tcagatacag gttccagccc agggccatct ggataaacag ggcgccgctg aagatatcgg
     3541 tagagatctt tgtaaagata tacaggatca gggacagcac gctcatgtac acctggattc
     3601 tctggccgcc gaaccgcttc ttcagatact gaggcattgt caccactcct gctgcgatgt
     3661 acacgggcac aaacacccat cccagggcca gcagcagcca ggttgcattc cactcgaatc
     3721 ctcccactgc cagtcctcct gctgctcctg tgcctgccag tccgataaac agtccgcttc
     3781 ccacgttgct agacatcaga gaggcgccga tgggccacca agacatggat ctgccggcca
     3841 ggaagtatcc tccgatggtt cccctgcttg ccctgatgga gctccagatg cccacggcga
     3901 tcacaaacac gaagtagatg accaccacag agatatcgta ggcgtgcagt cccactctgg
     3961 agtccagggc gatgtggggg gctgtctcgg tccgcacgcc gtcgccggat gctcctggtc
     4021 ccatggcggc cagctccttg ctcatggtgg agcctgcttt tttgtacaaa gttggcatta
     4081 taaaaaagca ttgctcatca atgtgttgca acgaacaggt cactatcagt caaaataaaa
     4141 tcattatttg gggcccgagc ttaagactgg ccgtcgtttt acaacgtcgt gactgggaaa
     4201 acatccatgc tagcgttaac gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag
     4261 gctcagtcgg aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
     4321 agtaggacaa atccgccggg agcggatttg aacgttgtga agcaacggcc cggagggtgg
     4381 cgggcaggac gcccgccata aactgccagg catcaaacta agcagaaggc catcctgacg
     4441 gatggccttt ttgcgtttct acaaactctt cctggctagc ggtacgcgta ttaattgcgt
     4501 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4561 gccaacgcgc ggggagaggc ggtttgcgta ttgggc
//
