LOCUS       sequence-461359-        3993 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 3993)
  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 3993)
  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-MFSD12_STOP
FEATURES             Location/Qualifiers
     source          1..3993
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      115..124
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1463..1562)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1578..1595)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1578..1594)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1587..1609)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      1658..1744
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1836..1863
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2022..2039)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2193..2781)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2273..2292)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2928..3737)
                     /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     3651..3670
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3729..3748
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     3850..3866
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3850..3866
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3871..3890
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3871..3889
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    join(3907..3993,1..13)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 aagaaagctg ggtcttatgg ccttgcatct ctgtcccacc ggcgcagcct tgtaggccac
       61 agcagcagag agcacaggca cagggcggct gccactccca ctcctccggt cactgccacc
      121 atggcccagt ggtagaagct cacgcaggcc ctacagcaca gctcagaggg gcatgggtgc
      181 agggactgga tggccatcac ggccaggcca ttggccacct tatccaggaa gctcatagag
      241 ccgtacacga aggcgccgga gtttgtgtgt ggtccgatca ggtctgcggt cattgccagg
      301 cttgtcacca ggatggttgc acatcctgct cccagcagca cggctgctgc gtacacggcc
      361 acgcccaggc cctcggccag ggccacccag gcggcgaagg ccaggatgac cagcagtccg
      421 ctgaagtatg tcatatttct gccgatgcac ttgttgatag gcttcatcag gaaagaggac
      481 agaaagccag acaggtacat gaccagtggg atggtggcga tgaacttctt aggcaggtgc
      541 aggctatatg tcaggtacat ggccatgtag gtctgagaca gattcacgat cagccttgtg
      601 gtcatataca ggatgcccac ttggtaaaag gcgggctctc tcagccagtg cttccacagc
      661 agcagtggct gtgctgttgc tggtgccagc agaggggtgt gctctcctgg ctcctcggcg
      721 tgtggccttc tccgctctct ggtgcccagg tggaacagca gggaaaacac ggctcccact
      781 cccaccacca gcaggctcag gttccggaac acgggcacat cctgtcctcc cagctgatcg
      841 gagatgctga tgtcctgggt tggctccacc ctgctagatc cctgcaggtg cagcagcagc
      901 catgctgctc catacactgt gatatttgcc accacggtga aggcgtatct cagggctgtc
      961 agctccacct tctcgtggtc gttggtcacc agctcgggga tcagagacag gtgggagatc
     1021 tgggtggagg cccagccgaa ctgaaagatc acgataaaag ggccatagta cagcagggcg
     1081 gcccactctg gtgttgctgc tccacagccc aggcaagggc tgaagataaa ggggaaagac
     1141 agcagcacgc acacggttcc caccaggtgc catgccttcc ttggtccata tcttgcacag
     1201 cagcttgctg ccctatctgc ctcgtatccc accagtggtg tgcacagtcc gtctgccacc
     1261 tgtcccagca gcagcagcag tcctgctccc ctggagctat atgctctcac agagtgcaga
     1321 tacagcagca ggtaggtgaa ccacatggag gcgcacaggt cgttcagaaa gtggcccact
     1381 gcgtagctca gccttgccac cagggacaga gggcgggggc ttggggctgc tcctgctgca
     1441 ggtggtccag gtcccatggt ggagcctgct tttttgtaca aagttggcat tataaaaaag
     1501 cattgctcat caatgtgttg caacgaacag gtcactatca gtcaaaataa aatcattatt
     1561 tggggcccga gcttaagact ggccgtcgtt ttacaacgtc gtgactggga aaacatccat
     1621 gctagcgtta acgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc
     1681 ggaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
     1741 aaatccgccg ggagcggatt tgaacgttgt gaagcaacgg cccggagggt ggcgggcagg
     1801 acgcccgcca taaactgcca ggcatcaaac taagcagaag gccatcctga cggatggcct
     1861 ttttgcgttt ctacaaactc ttcctggcta gcggtacgcg tattaattgc gttgcgctca
     1921 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     1981 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     2041 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     2101 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     2161 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2221 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2281 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2341 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2401 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2461 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     2521 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     2581 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     2641 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     2701 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     2761 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     2821 tggaacgacg cgtaactcac gttaagggat tttggtcatg agcttgcgcc gtcccgtcaa
     2881 gtcagcgtaa tgctctgcca gtgttacaac caattaacca attctgatta gaaaaactca
     2941 tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga
     3001 aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga
     3061 tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc
     3121 tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag
     3181 aatggcaaaa gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg
     3241 tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga
     3301 cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc
     3361 aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc
     3421 tggaatgctg tttttccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg
     3481 ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc
     3541 tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca
     3601 tcgggcttcc catacaagcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc
     3661 catttatacc catataaatc agcatccatg ttggaattta atcgcggcct cgacgtttcc
     3721 cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt
     3781 gttcatgatg atatattttt atcttgtgca atgtaacatc agagattttg agacacgggc
     3841 cagagctgcc aggaaacagc tatgaccatg taatacgact cactataggg gatatcagct
     3901 ggatggcaaa taatgatttt attttgactg atagtgacct gttcgttgca acaaattgat
     3961 aagcaatgct ttcttataat gccaactttg tac
//
