LOCUS       sequence-442087-        3903 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 3903)
  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 3903)
  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-MFSD5_STOP
FEATURES             Location/Qualifiers
     source          1..3903
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     77..94
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(253..280)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(372..458)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     507..529
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     521..538
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     522..538
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    554..653
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2013..2112)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2129..2148)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2130..2148)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2153..2169)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2153..2169)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2271..2290)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2282..3091
                     /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(2349..2368)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3238..3826
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3727..3746
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
       61 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
      121 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
      181 ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt accgctagcc aggaagagtt
      241 tgtagaaacg caaaaaggcc atccgtcagg atggccttct gcttagtttg atgcctggca
      301 gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc
      361 gctcccggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga
      421 aaggcccagt cttccgactg agcctttcgt tttatttgat gcctggcagt tccctactct
      481 cgcgttaacg ctagcatgga tgttttccca gtcacgacgt tgtaaaacga cggccagtct
      541 taagctcggg ccccaaataa tgattttatt ttgactgata gtgacctgtt cgttgcaaca
      601 cattgatgag caatgctttt ttataatgcc aactttgtac aaaaaagcag gctccaccat
      661 gctggtgacc gcatacctgg ccttcgtggg actgctggca tcttgcctgg gactggagct
      721 gagcaggtgt agagcaaagc cacctggaag ggcctgctcc aacccatctt ttctgagatt
      781 ccagctggac ttttaccagg tgtatttcct ggccctggca gcagattggc tgcaggcacc
      841 ctacctgtat aagctgtacc agcactacta ttttctggag ggccagatcg ccatcctgta
      901 cgtgtgcgga ctggcaagca cagtgctgtt cggactggtg gcaagctccc tggtggactg
      961 gctgggacgg aagaatagct gcgtgctgtt cagcctgacc tacagcctgt gctgtctgac
     1021 aaagctgtcc caggattatt tcgtgctgct ggtgggaaga gccctgggag gactgagcac
     1081 cgccctgctg ttttccgcct tcgaggcctg gtacatccac gagcacgtgg agaggcacga
     1141 ctttccagcc gagtggatcc cagccacatt tgccagggca gcattctgga accacgtgct
     1201 ggcagtggtg gcaggagtgg cagcagaggc cgtggcatct tggatcggac tgggacctgt
     1261 ggcaccattc gtggccgcca tccctctgct ggccctggca ggcgccctgg ccctgaggaa
     1321 ctggggcgag aattatgaca ggcagcgcgc ctttagcagg acctgcgcag gaggactgag
     1381 gtgtctgctg tccgatcgga gagtgctgct gctgggcacc atccaggccc tgttcgagtc
     1441 cgtgatcttc atcttcgtgt tcctgtggac accagtgctg gatccacacg gagcacctct
     1501 gggaatcatc ttttctagct tcatggccgc ctctctgctg ggctcctctc tgtacaggat
     1561 cgccacaagc aagcggtacc acctgcagcc tatgcacctg ctgtccctgg ccgtgctgat
     1621 cgtggtgttt tctctgttca tgctgacctt tagcacatcc cctggccagg agtccccagt
     1681 ggagtctttt atcgccttcc tgctgatcga gctggcctgt ggcctgtact ttccctctat
     1741 gagcttcctg aggcgcaaag tgatccctga gaccgagcag gcaggcgtgc tgaactggtt
     1801 cagggtgcca ctgcactccc tggcatgcct gggactgctg gtgctgcacg actctgatcg
     1861 gaagaccggc acaagaaata tgtttagcat ctgtagcgcc gtgatggtca tggcactgct
     1921 ggctgtcgta gggcttttca cggtagttcg gcacgacgca gagctgagag tgccatcccc
     1981 cacagaggag ccctatgccc ctgagctgta agacccagct ttcttgtaca aagttggcat
     2041 tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa
     2101 aatcattatt tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc
     2161 tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     2221 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
     2281 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
     2341 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     2401 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     2461 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     2521 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     2581 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     2641 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     2701 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     2761 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     2821 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     2881 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     2941 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     3001 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     3061 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg
     3121 gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt
     3181 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3241 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3301 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3361 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     3421 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3481 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3541 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3601 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3661 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3721 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3781 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3841 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     3901 atc
//
