LOCUS       sequence-312050-        4218 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 4218)
  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 4218)
  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-FLVCR1_STOP
FEATURES             Location/Qualifiers
     source          1..4218
                     /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"
     protein_bind    complement(2343..2442)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2459..2478)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2460..2478)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2483..2499)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2483..2499)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2601..2620)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2612..3421
                     /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(2679..2698)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3568..4156
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4057..4076
                     /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 accatggcca ggcccgacga tgaggaggga gcagcagtgg cacctggaca
      721 cccactggcc aagggctatc tgccactgcc taggggagca cctgtgggca aggagagcgt
      781 ggagctgcag aatggcccaa aggcaggcac cttccctgtg aacggagcac caagggactc
      841 cctggcagca gcatctggcg tgctgggagg accccagaca cctctggccc cagaggagga
      901 gacccaggca aggctgctgc cagcaggagc aggagccgag acacctggag cagagagctc
      961 cccactgcca ctgaccgccc tgagccctcg gagattcgtg gtgctgctga tcttttctct
     1021 gtacagcctg gtgaatgcct ttcagtggat ccagtattcc atcatctcta acgtgttcga
     1081 gggcttttac ggcgtgaccc tgctgcacat cgactggctg tccatggtgt acatgctggc
     1141 ctatgtgccc ctgatcttcc ctgccacctg gctgctggat acaaggggac tgagactgac
     1201 agccctgctg ggatctggcc tgaattgcct gggcgcctgg atcaagtgtg gcagcgtgca
     1261 gcagcacctg ttttgggtga ccatgctggg acagtgcctg tgctccgtgg cacaggtgtt
     1321 catcctggga ctgccaagca ggatcgcatc cgtgtggttt ggaccaaagg aggtgtctac
     1381 cgcatgcgca acagccgtgc tgggaaatca gctgggaaca gcagtgggct tcctgctgcc
     1441 acctgtgctg gtgcccaaca cccagaatga cacaaacctg ctggcctgta acatcagcac
     1501 catgttttat ggcacatccg ccgtggccac cctgctgttc atcctgaccg ccatcgcctt
     1561 taaggagaag ccaaggtacc caccatctca ggcacaggcc gccctgcagg atagccctcc
     1621 agaggagtac tcttataaga agagcatcag aaatctgttc aagaacatcc cctttgtgct
     1681 gctgctgatc acatacggca tcatgaccgg cgccttctat tccgtgtcta cactgctgaa
     1741 tcagatgatc ctgacctact atgagggaga ggaggtgaac gcaggaagga tcggactgac
     1801 cctggtggtg gcaggaatgg tgggaagcat cctgtgcggc ctgtggctgg actacaccaa
     1861 gacatataag cagaccacac tgatcgtgta tatcctgtcc ttcatcggca tggtcatctt
     1921 caccttcacc ctggatctga gatacatcat catcgtgttt gtgacaggcg gcgtgctggg
     1981 cttctttatg accggctatc tgcccctggg cttcgagttt gccgtggaga tcacataccc
     2041 tgagtccgag ggcacctcta gcggcctgct gaatgcctct gcccagatct tcggcatcct
     2101 gtttacactg gcccagggca agctgacctc tgattacggc cctaaggccg gcaacatctt
     2161 cctgtgcgtg tggatgttta tcggcatcat cctgacagcc ctgatcaagt ccgacctgag
     2221 gcgccacaac atcaatatcg gcatcaccaa cgtggatgtg aaggccatcc cagccgacag
     2281 ccccacagat caggagccaa agaccgtgat gctgtccaag cagagcgagt ccgccatcta
     2341 agacccagct ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg
     2401 caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc
     2461 ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca
     2521 aaatctctga tgttacattg cacaagataa aataatatca tcatgaacaa taaaactgtc
     2581 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag
     2641 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     2701 tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt
     2761 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     2821 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
     2881 tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga
     2941 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     3001 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
     3061 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     3121 ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc
     3181 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
     3241 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     3301 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     3361 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     3421 atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg
     3481 cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga
     3541 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3601 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3661 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3721 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3781 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3841 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3901 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3961 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4021 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4081 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4141 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4201 gccttttgct cacatgtt
//
