LOCUS       sequence-479324-        5502 bp    DNA     circular SYN 05-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 5502)
  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 5502)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 5, 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-SLC4A9_STOP
FEATURES             Location/Qualifiers
     source          1..5502
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(25..42)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(196..784)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(276..295)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(931..1740)
                     /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     1654..1673
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1732..1751
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1853..1869
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1853..1869
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1874..1893
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1874..1892
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1910..2009
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(4968..5067)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(5083..5100)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5083..5099)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5092..5114)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      5163..5249
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5341..5368
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
       61 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
      121 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      181 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      241 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      301 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      361 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
      421 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
      481 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
      541 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
      601 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
      661 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
      721 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
      781 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg acgcgtaact
      841 cacgttaagg gattttggtc atgagcttgc gccgtcccgt caagtcagcg taatgctctg
      901 ccagtgttac aaccaattaa ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa
      961 ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa
     1021 tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc
     1081 gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt
     1141 atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagtttatg
     1201 catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc
     1261 atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct
     1321 gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc
     1381 atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg ctgtttttcc
     1441 ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt
     1501 cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt
     1561 ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa
     1621 gcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa
     1681 atcagcatcc atgttggaat ttaatcgcgg cctcgacgtt tcccgttgaa tatggctcat
     1741 aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt
     1801 tttatcttgt gcaatgtaac atcagagatt ttgagacacg ggccagagct gccaggaaac
     1861 agctatgacc atgtaatacg actcactata ggggatatca gctggatggc aaataatgat
     1921 tttattttga ctgatagtga cctgttcgtt gcaacaaatt gataagcaat gctttcttat
     1981 aatgccaact ttgtacaaga aagctgggtc ttaattcacg ctgatgttga tctcaggggc
     2041 cttgggctgg tacatcagct cagaatcctc agagtcggag ccgctaaaag agtgctcagg
     2101 ctccagtccc ttctctggga tgctccgctc ctcctcgggc atcagctcat ccagccacag
     2161 cagctcctgt ggggaaaaca cgcgctccag ggccttcctc acgcccacca gtcccagcag
     2221 catcagtggg aagatgatgg cggcaggtgt gctcttgatg atccacagca gtcccaggca
     2281 tgccagctgg attgctgtga acaggtgcac tctggtcagt ggcacgtgcc tcagcagcag
     2341 caggtctggc tggtgcttgg ctggcatcag cagcagcttc accctattgg tgaactggat
     2401 gctagacagg gcggccacgc ccatatacag aaagatgccg tacagcactg gcatagggat
     2461 gaacttcagc acaggggcca gaaagatgga ggcgcctgtc aggataaaca ccaccaggcc
     2521 ggtcagtctc tgctcccgga ttcccaggaa gttaggcctc tctcctggtg cacatgccct
     2581 gctctctctc cgcagagagt ccatgtgggc cagggagatc actgtggcgc tcacatacca
     2641 aggcagtccc agggcgctgg tcagcagcat cagcacggcc acgcagaaca gatccaggtg
     2701 aaagccggcg cccttctgca gtctgtactc catccgattc aggatcacgg cggtgatctg
     2761 ctggtccatg aagatcagga tggacagcag cagggcgggc agggcggctg ccacgctcca
     2821 ccaccaaggg ttggcgccaa agggagacac cagccatccc cttcctggca gtgtaggctt
     2881 gaactccctt ggcaccatca gcttaggggt ggccaggccc agaaaggcat ccagtccaca
     2941 tcccagcagg attgccagca cggagctaaa gtcgctcagg ccctttctca ccacagatgg
     3001 gaaaaaccgg gatgtcttca cgcacttcag ggccatggcg aaaaagaaag aggtcaggaa
     3061 cagcagcagg gaaaagaagg cgatatcagg cactgtgtga cagcctggtc cccttgggtg
     3121 tcctccctgc ctggtgcact ctggtggagg cagcagggat gcattgatca ggcccagatc
     3181 catgctcacg atatcgtctc tgtccttggg gcgggtcctg atccactggc tctcgtttcc
     3241 tcctggtcct ggatactggc acaggcatcc gtaggcagag gagcctggct tctggatagg
     3301 atatgtgtgg gtcagattca gcatcttgcc cacggcgtcg tagatgaaga tcagggagat
     3361 cagggcacaa aagccctcct ctgtgaagcg ggtgaagtac ctcaccagca cgcttgcctc
     3421 tgttgccacc agcaccaggc agaaggtggc cacccagatg cccacccaca gcctaaaggg
     3481 cagataatcc aggctgtagt cgcgggagaa gctaaacagc agcctctcga acaccagcac
     3541 ggggcctgtg ctagacagga tggtcagtgg ctggccggcc atcagacaga atgctgctcc
     3601 tgccactgcg gttcccagaa aggactccag cacgccctgg gctccatctg ttgcgtcgcc
     3661 cagcagtccg ccaaaggtga tggcgtttgt cacggtggcc agatagatgt acagcacggc
     3721 agagaagcac tgcaggtgca gggcatccag aaagtcggaa gggtaccagg gcaccttgcg
     3781 cctcacatcc tggatcagtc ctccgaacag gcggcctgtc ctctgcagct ctggggagtg
     3841 tgcgtgtggt ccgtgtctgt gcctgtcctc ggcgctggtc agccttggca ctgcaggtcc
     3901 tctgatctcc ctctgctggc ttggcagcct cttgtgctga gaaggcagac acttgggtgg
     3961 agggattctg gctgtgggat cccaccggcc tggtggcagc acggtcacct cctccagaaa
     4021 ggcatccagg gcggccagca ggtcgtgcag attgctggct ctccgcacag accactggaa
     4081 ctgtgggtca gacagcagca cggctgctgc tcttcccatc tcgtgatagc ccttgcccag
     4141 catacaaggg cccagcagca ggcaaaagaa ccgggagggc aggctcacct ctgtcaggga
     4201 tcccagcacc actgggttcc tcagcctcac gaaggctccc agaggctgtg ccagaaagcc
     4261 cagctctcct gccagcacgg ttcctgcctc tgctccaggg ggcagcttct gtctcagagg
     4321 attctgacac tgctccctca gtggtgcctc ctcgttatcg gatgcctttc tggggtgtgt
     4381 gcttccccag catggcctgg ttcctgtggt ctgattgtag tgctgtgggc gctgcagcag
     4441 cagggcctgc agctgtcccc tcagctcagg agacagggac tccacccttg tcacctgctc
     4501 caccagctcc agcaggctct gtgcagggca gtccagcagc accagtccct ctgccagcag
     4561 agacctcagc ttctgcaggg atggcagggc cagggtgggc acgtgtgggg cagaccatct
     4621 gcctgctgcc acctccagct tctcctcgaa cagcacccac cggtgatgca ggtgtgtaga
     4681 cagggtgatg gagggcatct cgcccatatc cagcagcagg ctggcggagc tagatccggt
     4741 ctctctccac tccagggcct gtggccatcc cagcagctcg ttcagctgga taaacagcag
     4801 agggtccttg ggcacgccca gctccttgct ctctgtatca gaaggtccgt caggggatgg
     4861 tccggttcct ggatcaggat tggagtccag ctctccgctt gggatgttcc ttggggcgga
     4921 gcttgcctcg aatccctcct gtcctggcag cttcatctcc atggtggagc ctgctttttt
     4981 gtacaaagtt ggcattataa aaaagcattg ctcatcaatg tgttgcaacg aacaggtcac
     5041 tatcagtcaa aataaaatca ttatttgggg cccgagctta agactggccg tcgttttaca
     5101 acgtcgtgac tgggaaaaca tccatgctag cgttaacgcg agagtaggga actgccaggc
     5161 atcaaataaa acgaaaggct cagtcggaag actgggcctt tcgttttatc tgttgtttgt
     5221 cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgtgaagc
     5281 aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat caaactaagc
     5341 agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttcct ggctagcggt
     5401 acgcgtatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5461 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt tt
//
