LOCUS       sequence-442079-        4095 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 4095)
  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 4095)
  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-MFSD4A_STOP
FEATURES             Location/Qualifiers
     source          1..4095
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     187..206
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     440..457
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(616..643)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(735..821)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     870..892
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     884..901
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     885..901
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    917..1016
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      1428..1437
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2568..2667)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2684..2703)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2685..2703)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2708..2724)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2708..2724)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2826..2845)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2837..3646
                     /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(2904..2923)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(3793..4095,1..286)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
       61 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
      121 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
      181 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
      241 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
      301 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
      361 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
      421 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
      481 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
      541 cgactggaaa gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta gccaggaaga
      601 gtttgtagaa acgcaaaaag gccatccgtc aggatggcct tctgcttagt ttgatgcctg
      661 gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac aacgttcaaa
      721 tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca acagataaaa
      781 cgaaaggccc agtcttccga ctgagccttt cgttttattt gatgcctggc agttccctac
      841 tctcgcgtta acgctagcat ggatgttttc ccagtcacga cgttgtaaaa cgacggccag
      901 tcttaagctc gggccccaaa taatgatttt attttgactg atagtgacct gttcgttgca
      961 acaaattgat aagcaatgct ttcttataat gccaactttg tacaagaaag ctgggtctta
     1021 gcgctgataa cactcgctat tctccatgcc gatagaccta tcctgtgttg gcacagaagg
     1081 caggccgggg tgcattctgt gaaagaacag cagcaggata taaaaggtga aggccagaca
     1141 gccaaagatc acgccgcaca ccaggaaaga gtaggagccc tgggcctgga agatggagcc
     1201 caccagcatc tgcagcacca tctctcccac tcctgctcct gtcaccagca ctgtggttgc
     1261 gcagccctta tactgcagag agtcctctgt gtaggccagc atggagggaa aggtgctaga
     1321 caggaacagt cccaggcttg cggttcccac aaacaggaac accacattgt aagaaaagat
     1381 cagcagcacc aggaatgtca ccaccacgcc caccacgttg atgaacacca tggtggcagg
     1441 cttcattctg gagctgatgg ggatggacag cagccggccc agggtgataa agccccagaa
     1501 cagagagggc agatagcctg ccaccttgtg tcccacggac agtggcttct ccacggcata
     1561 agagtacacg aaggcgctgt aggcgcctgt caggccatcg gtcatgaaca gcaccagggc
     1621 gccggtgatg tggatggcaa agaagctata aggggcgcct ctcaggttct tccgctgaca
     1681 gcagctaaac aggtcctcgt ggcccaggtg agactggaac ttgggtggca gagaggaggc
     1741 atcctccttc tcagggggct gtgtctccag ggccagctcg tcggcggaca gcagcagggg
     1801 gcgcctctgt ggacagcagg tcagcaggcg ctccttgctc agcagcatca gcacggccat
     1861 tggcacaggc agattgatca gggccatgat ccaaaaggcg tagctcaccc ttgtgcctgc
     1921 tccatcctta ggtgtcagtc ctgggaaggt ctggttagac caaggctttg cgtccacgtg
     1981 gtgctgtccc agcacgcggg acacgtgaaa caggtggccc ctggatgtgg tattggcggt
     2041 gctgttggcg ggcaggcaat tggcctcgga caggaatgga tcggcgatca gagggctcag
     2101 cagggcgcca aagcccacaa agaagtgcag cacctgcagg aacacggcgc tatccttctg
     2161 atacatcctc accagctgca tgttggccac ggtgtcgata catcccattg ccagtcctgc
     2221 cagtgccatc acagatgcca gcaccttcac gtctctgcaa aaggggatca cggcgaacac
     2281 cagggagatg gccaggctag atgtaaacag tgcccacagg ctctgtgcca gggtccgctt
     2341 gaacacgcct cccagggcgg atcccagcag caggcacagc tgctggctaa agaacaccca
     2401 agagatctgt ggcagggagc tgtgtgtctg acatctcaga tccagcaggg tagggcccag
     2461 aaaggcgatg cacaggccga aggaaaagaa cacgctccag tatgtcaggg ttggctgcag
     2521 gtttctccgc agcagtccgc tcacccgtcc gtcgcatccc atggtggagc ctgctttttt
     2581 gtacaaagtt ggcattataa aaaagcattg ctcatcaatg tgttgcaacg aacaggtcac
     2641 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta
     2701 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     2761 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     2821 taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc
     2881 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
     2941 tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg
     3001 caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga
     3061 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     3121 caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     3181 tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     3241 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     3301 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     3361 agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg
     3421 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     3481 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     3541 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     3601 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa
     3661 ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc
     3721 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     3781 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3841 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3901 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3961 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4021 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4081 gttaccggat aaggc
//
