LOCUS       sequence-258985-        4308 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4308)
  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 4308)
  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_MFSD6L
FEATURES             Location/Qualifiers
     source          1..4308
                     /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(2433..2532)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2549..2568)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2550..2568)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2573..2589)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2573..2589)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2691..2710)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2702..3511
                     /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(2769..2788)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3658..4246
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4147..4166
                     /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 accatgagcg ccaaccctcg gtgggatatc tccagagccc tgggcgtggc
      721 caagctgttc cacctggtgt gcggcgtgcg ggaggcatgc gtgacccctt ttctgacact
      781 gtacctgaga cagctgggac tggcagcacc atgggtgggc accctgatgg gcacaaagca
      841 cctgatcgca gcattctggg cacccgtgtg cgcctttctg gccaagagct ataggaagcg
      901 gagagccctg ctgatcggat ctctgctggg aagcgtggga gcatccctgc tgatggtgct
      961 ggtgccccct gtggataaga accgcgtgca cttcccatgt aatggaagct ccggactgac
     1021 ctccacagac gcactgcctg gagtgaccct gccagtgaac atcacatctg cccaggagag
     1081 cgcctctagc cacccagcaa agaggaccgc agaggtggag atgccaggct tccgcaatcc
     1141 accaggagag agcgataggg agaccttccg ggacctgcac gtgtacctgg caccatctgt
     1201 ggagggagca aggaccacaa gccaggccct gctgcaccca gtgacctccg gcctgaagga
     1261 tcacccttgg gaggtgacct tcgaggtggt gaagacagcc ctgccactgc tgcctggagg
     1321 caagggacca ggaaacccag caaatctgtc cggcacaaag ggcaaggcct gggcctttga
     1381 cctgtctctg gaggccctga ggcgcacctt catcctgagc ctgggctccg tggccttttg
     1441 ggagctgctg acagcaccac tggagcaggt ggcagacgat agcctgtacg agttcctgga
     1501 ctttgtggat gccaccgaca ggtatcgcag cctgtgggtg tggaggctgc tgggaatgtc
     1561 cgccggcgtg tgcggaatca ccgccctggt gggccagctg gattgtttcc tgatgacatc
     1621 cggccctaga ggcgtggtgc acttttacgg ctattctgtg gtgagcacac tggccctgct
     1681 ggtgtctatc gccttcccta tcccaatctg ccagcagtgg gagccctcct ataagagagt
     1741 gaaggccctg tctatcgtgg gcggcgatcc tcacctgatc ctgctggcct ctaccacagt
     1801 gctggtgggc gccatcgtga gcaccgtgca gaacttcctg ttttggcaca tgaaggacca
     1861 cggatccgga gagctggtca tgggcttctc cgtggccctg tctctgctgg gcgagatcct
     1921 gctgcacccc ttcaaggcca ccctgctgag gaagctgtcc aggacaggac tggtgggact
     1981 gggactgtct tgcctggcag gccagctgct gtactatagc ttcctgtggt cctggtggtc
     2041 tgtgctgcca atccagatcc tgagcgccat ctccaatcgg gccctgtggt gggcagtggg
     2101 agcaagcgtg gaggacctgg caaccccaag gatggagcgc gccctgtccg ccctgttcag
     2161 gggccacttt tacggctctg gatgtagcct gggatccttc gtgggaggat ttgtggtcat
     2221 gcgcttttct ctggccgtgc tgtatcaggc ctgctgcgtg gccctgctgc tgtggctggc
     2281 cctgctgctg agcatccagc ggagactgcc ccgggagaga aagatcaagt acagcaagct
     2341 gctgtccatg gaggtgtctg atacctccga ctctgagcag ggcacagagc aggattggct
     2401 ggtgaaggcc atgagagagg agcacagcga cgacccagct ttcttgtaca aagttggcat
     2461 tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa
     2521 aatcattatt tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc
     2581 tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     2641 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
     2701 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
     2761 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     2821 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     2881 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     2941 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     3001 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     3061 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     3121 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     3181 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     3241 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     3301 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     3361 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     3421 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     3481 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg
     3541 gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt
     3601 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3661 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3721 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3781 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     3841 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3901 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3961 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4021 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4081 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4141 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4201 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4261 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtt
//
