LOCUS       sequence-312006-        4311 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 4311)
  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 4311)
  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_STOP
FEATURES             Location/Qualifiers
     source          1..4311
                     /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(2436..2535)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2552..2571)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2553..2571)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2576..2592)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2576..2592)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2694..2713)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2705..3514
                     /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(2772..2791)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3661..4249
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4150..4169
                     /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 ctaagaccca gctttcttgt acaaagttgg
     2461 cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa
     2521 taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat
     2581 agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga
     2641 taaaataata tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg
     2701 tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc
     2761 tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta
     2821 tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt
     2881 tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct
     2941 tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat
     3001 ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt
     3061 tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt
     3121 taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt
     3181 tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga
     3241 aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact
     3301 tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg
     3361 aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc
     3421 ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt
     3481 gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca
     3541 ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa
     3601 cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3661 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     3721 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3781 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     3841 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3901 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3961 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4021 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     4081 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4141 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4201 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     4261 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt t
//
