LOCUS       sequence-461352-        4071 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 4071)
  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 4071)
  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-MFSD14B_STOP
FEATURES             Location/Qualifiers
     source          1..4071
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1579..1678)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1694..1711)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1694..1710)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1703..1725)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      1774..1860
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1952..1979
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2138..2155)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2309..2897)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2389..2408)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3044..3853)
                     /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     3767..3786
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3845..3864
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     3966..3982
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3966..3982
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3987..4006
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3987..4005
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    join(4023..4071,1..51)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 ttgataagca atgctttctt ataatgccaa ctttgtacaa gaaagctggg tcttacagct
       61 cggtacactg attgccgggc tcctcgaagc tagacagctc ccagatggag ctatcctgca
      121 gcagtggctc gatatcctcg tcgctgcctc tctcaggtgt attggtcaga gagccagagg
      181 agctgttgga gtgcttctgc acgccagagg ccttggagta ctctgggatg aacagggcca
      241 ccagaaaaga catcagcacg atacatgctc caaacaggaa aggtggtcct gggatcacgg
      301 ctccctgcag tggcacattg ttggagttca gctttgggcc cagctcggtc agctccacgt
      361 ggaacatata aaagatgaag ccgtacaggg caggtcccag tccattgcac agtcccctga
      421 ttcctgtgat gatgccctgt gccactccct gctggtcgct ctctgcgttc ctgctcacca
      481 gggcagagat ggcgggaaag gtgatagagg acattgctgc cactgttcct gctgcccaca
      541 tcatccaggc ctgagagccg aagccatacc aggccagctg cagcatctga aagcccaggc
      601 ccagcagcac ggtattcttg ttgcccagag agcgcatcag gatggacagg aaggctgtct
      661 gggccacgat ggacaggatg cccaccatgg cgatgaaggc ggcgatcttc acgctgccaa
      721 agccgatgac ctgcctcaga tacagaaaga agctagagta ctggccggcc tcaggcaggt
      781 aggacaggaa cacggtgata cagatcagca gcactgtgga atccttgccg accttcttca
      841 ggctggcaaa ggggtctgcc tgcttccagc tgatctgtgc tccccaagac actggccgca
      901 tcttctctgg caggctctca ggcacggcca ccaggatgaa gcagatatcc agcagggcca
      961 ccacggttgc caccagcacc accagagagt cgccatagga ggcgctcagg tatgctccga
     1021 ttgctgggga gctcaccagg gatgcggcaa aggtggcgct cacccatcca tatgcggtgc
     1081 tccgctcgtg ctcctgtgtc acatcggcca cgtaggcgaa gatcacggaa aatgtcacgc
     1141 tgaacacgcc agacacggag atcatggcga aataccacca aggggagatt ctcatcaggg
     1201 ggattgggaa gcatgtaaag aacacggtgc ccagcagaaa aggcttccgg ccccacacgt
     1261 cgctcagggc gccgatcagt ggggcgctca gaaaagacag caggcccttc acgccctgga
     1321 tcaggccgtt catcaggaag gtgtgctggg aaaatgtctc gtgcagcacg gtcagcatag
     1381 gtgtggtcag caggccccag gcaaagaact ccaggaagat cacgatggcg gcgtggtaca
     1441 cgctgggtct gccaaagccc tgcagccatg tagatcctgc tgcctgtgct cctgccctct
     1501 tctcaggcat ggctcctgcc tctggctcag atgctgcctt ctcctccagc tctggaggtg
     1561 gctccacgga catggtggag cctgcttttt tgtacaaagt tggcattata aaaaagcatt
     1621 gctcatcaat gtgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttggg
     1681 gcccgagctt aagactggcc gtcgttttac aacgtcgtga ctgggaaaac atccatgcta
     1741 gcgttaacgc gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcggaa
     1801 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat
     1861 ccgccgggag cggatttgaa cgttgtgaag caacggcccg gagggtggcg ggcaggacgc
     1921 ccgccataaa ctgccaggca tcaaactaag cagaaggcca tcctgacgga tggccttttt
     1981 gcgtttctac aaactcttcc tggctagcgg tacgcgtatt aattgcgttg cgctcactgc
     2041 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     2101 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     2161 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     2221 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     2281 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2341 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2401 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2461 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     2521 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2581 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     2641 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2701 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     2761 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2821 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2881 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     2941 acgacgcgta actcacgtta agggattttg gtcatgagct tgcgccgtcc cgtcaagtca
     3001 gcgtaatgct ctgccagtgt tacaaccaat taaccaattc tgattagaaa aactcatcga
     3061 gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
     3121 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
     3181 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
     3241 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
     3301 gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
     3361 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa
     3421 atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
     3481 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
     3541 atgctgtttt tccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
     3601 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat
     3661 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
     3721 gcttcccata caagcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
     3781 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctcgac gtttcccgtt
     3841 gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc
     3901 atgatgatat atttttatct tgtgcaatgt aacatcagag attttgagac acgggccaga
     3961 gctgccagga aacagctatg accatgtaat acgactcact ataggggata tcagctggat
     4021 ggcaaataat gattttattt tgactgatag tgacctgttc gttgcaacaa a
//
