LOCUS       sequence-479316-        5925 bp    DNA     circular SYN 29-JUL-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 5925)
  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 5925)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 29, 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-SLC9C2_STOP
FEATURES             Location/Qualifiers
     source          1..5925
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(26..43)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(197..785)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(277..296)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(932..1741)
                     /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     1655..1674
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1733..1752
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1854..1870
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1854..1870
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1875..1894
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1875..1893
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1911..2010
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(5392..5491)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(5507..5524)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5507..5523)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5516..5538)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      5587..5673
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5765..5792
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
       61 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
      121 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
      181 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
      241 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
      301 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
      361 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
      421 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
      481 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
      541 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
      601 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
      661 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
      721 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
      781 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gacgcgtaac
      841 tcacgttaag ggattttggt catgagcttg cgccgtcccg tcaagtcagc gtaatgctct
      901 gccagtgtta caaccaatta accaattctg attagaaaaa ctcatcgagc atcaaatgaa
      961 actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta
     1021 atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg
     1081 cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt
     1141 tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat
     1201 gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg
     1261 catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc
     1321 tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg
     1381 catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat gctgtttttc
     1441 cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg
     1501 tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat
     1561 tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca
     1621 agcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata
     1681 aatcagcatc catgttggaa tttaatcgcg gcctcgacgt ttcccgttga atatggctca
     1741 taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat gatgatatat
     1801 ttttatcttg tgcaatgtaa catcagagat tttgagacac gggccagagc tgccaggaaa
     1861 cagctatgac catgtaatac gactcactat aggggatatc agctggatgg caaataatga
     1921 ttttattttg actgatagtg acctgttcgt tgcaacaaat tgataagcaa tgctttctta
     1981 taatgccaac tttgtacaag aaagctgggt cttagctcat cttctgcctg ccgttgatat
     2041 tcttgccggg ctgctcgaac actgtgttca cgctggccat cacgttggta ttagagtttc
     2101 tctgtgtcag ctcggaggcc tggatgatca gcagcttgga cagatcgctg gtgccctgca
     2161 cctgctcaca tgtggtaggg atgatgcagg gggcgaaata tggctcctct gtcttggtgt
     2221 cgatcacgga gccgtacacg atgatcacga acttcattgt catgttatcg atgatcatgt
     2281 cgctataaga ggacagggtc tccacgtagg cctggttgaa catcacgcaa ttctgaaagc
     2341 gcagatcctc gtcgatcagg ctagactcga aatactggta ggcggtagac agggccagct
     2401 tcagccagat cttatactcc agggatggcc aaaaggcatc gaagccctcg tacaggtcct
     2461 ccaggctgat aaaacaggcc tgcagagagg tctcgcagat cactgtgtac tcgatctccc
     2521 tcttcagcag acagctcagc tcgccgatga tgtcgcctgt ggtgcaaaac tctgtgaaca
     2581 tatccctaga tcccctgtcg cacctctggt tggactcgat gccgaaggtg ggagacaggg
     2641 agtgcaggat ggccatgcca gagatgatca gatagatgcc ctgtggcatc tcgccgccct
     2701 tacagatggt atcgccggag tcaaagcagg ccagcttggc tctctcctta aagaaatcga
     2761 tcagcacgtc cttgccctcc agccagatga tgttgtgcag gtagatatcg ggggtagggg
     2821 gtgggatggc ctttgggaaa ttgttcaggg ccttcagctt cttcagcagc accttattga
     2881 tctcgatcac ttcgtgcttg tcgatgatgc cccgggagca cagaaatgtc aggttcttca
     2941 gggccttggc gatcacattc cggatggcct gcttggtctt cagggcgatg accacgtccc
     3001 ttccctcgtg ctccatcagc accagctcct tcacggcgtc ctgcttattt gtctccagga
     3061 tctcgcacag cttctggtag atagactcgc acacggcgat ctgcttgatc agcagcttgg
     3121 catcctcctg ggacttgata tagcccttgg tgatagagta catcagggac agcctcttct
     3181 tgatctgcac gtcggcgatt ctgatcagga taggcacgat gatcttaaac aggggcagga
     3241 atctgatgat ccgcagatag cccatgatca cggtcagctg gatcagggcc aggttatcgg
     3301 ggcgcagctt cacaaagtac acgcagaaga tgtcgatgat gccgatgacc aggataaaga
     3361 actccagtgt attccagcac tgctggaaat acttcctctt caggatgatg atcttcaggg
     3421 tgctctccag cacgtacaga aacatgaaat agtagttgat gctgatcagg gcgctcacgt
     3481 tcaggcctct ggccattggc cacaggtgga tgatcatagg atagatgtag atcaggttga
     3541 tgatctggcc tgtatactca aactcctcgg agaacacgat gtgaaagatg aatgtcagga
     3601 aggtgttgct ctcagggggg ataaagtgga tcttctcgat acagtactcc aggaaggtca
     3661 gcacattctt aaacttgatc agccagcttc ttgtccgcat ataggtgctc acgtcgtaga
     3721 tgctcatgaa cttgccctgg atagaatagt agcacttggc tgctccgatc aggatccttg
     3781 ctgcctcgat ctccaggatg ccattgtttc tctgcttctc aaaggagctc atctggatgg
     3841 ctgccacgtg cagccttgcc tcctccatca gggcctcatc tgtggtggac tctgtcttca
     3901 tgtcgttgtg agacacgtgg gagaaaggga tgtactcgat gcgggtcttg tcctccacca
     3961 gggtccaatt cacgtttgtc aggatcttct cggtcttaaa cagtgtgatg gtgttctgca
     4021 cgatctcctg gatgtgctgg gtggcattct gcaggatcat ctgccggggc agggacagca
     4081 cgcacagatc cagcttgcgg gcggactggg tcatcacata gctattgatg cccattgtca
     4141 gcaggctgat gacctgcacg tacaggatga acatctgtgg cacctccacc ttccgctcgg
     4201 ccaggttgta cacgtcaggg gcccacagca gattaaacac gcccttgatg ccgctccagg
     4261 tgatgaccac tccccatctc cagttatact cgtaattaga gtgcatcagg attggggaca
     4321 ccagcaggat tgtcagcagg cgcaccaggt tcactgtggt gaacaggata aagatgaaag
     4381 ggatggtgtg aaactcgtag tgagacagct cgccacagcc gatcacgatg ccaaagaagg
     4441 catagatcag gtgctcgtac acgctggaaa agatgcgcag gaacttggtg atgaccagct
     4501 cgatcttggg cttgaatgtc aggctatcca gattcaggcc cactgctgcc agggccagtg
     4561 tgccagacat tcccagaaac tccacgatat agaaggtcat gtacaccatg ctgaagcaca
     4621 ggatgatgtt tgtcagcata ttagaaaaca cgtcggccag gatacactgg atgatcttgg
     4681 cgcaccaata gccgaagatg atgctgccca ggatatcgta agacagctcg atgcccacgt
     4741 gcaggtccct aaagatgctg aagtggatgc ggttgccccg gaagttgcca aagaagatgc
     4801 tggcgataga acagatgatc agggactcgc ccctgatcag atcgatgtag atcttgctga
     4861 tgccgattgt cttcagggag ttcacgcttc tcagagggtc gatgatgccc agggtgatag
     4921 agaacagcag gcaggactgc agatcccagc tgtccttgtt gaacttgatg accacatagc
     4981 cgatgatgat gctggcggtg ctgaaagaga tcaggcctgt cagcagcacc tgccagaaca
     5041 tcttcttcag tgtgtaaaac tccacgtcca gggccaccat gaagatgatc agaggagaaa
     5101 aataggagta caggctgaag ctagaggttc tcagcagggg atacacgatc tggtgcacct
     5161 ccacggagtt gtaggccatg tggccgatca cgaagccgga caggctcagg atggtcagca
     5221 cgatcacttc acaattcttc aggcacatct tcagcaggcc gcccagcacc acgataaagc
     5281 acaccagggt ggtgaagtgc ttctcctcca ccagataatc ggctggctgt ccgcacagca
     5341 ggtctggcct gttggactca ttctgggccc aaaagtagga gctcatggtg gagcctgctt
     5401 ttttgtacaa agttggcatt ataaaaaagc attgctcatc aatgtgttgc aacgaacagg
     5461 tcactatcag tcaaaataaa atcattattt ggggcccgag cttaagactg gccgtcgttt
     5521 tacaacgtcg tgactgggaa aacatccatg ctagcgttaa cgcgagagta gggaactgcc
     5581 aggcatcaaa taaaacgaaa ggctcagtcg gaagactggg cctttcgttt tatctgttgt
     5641 ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgtg
     5701 aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaact
     5761 aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct tcctggctag
     5821 cggtacgcgt attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     5881 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtt
//
