LOCUS       sequence-259556-        3990 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 3990)
  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 3990)
  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
FEATURES             Location/Qualifiers
     source          1..3990
                     /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"
     regulatory      2007..2016
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2115..2214)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2231..2250)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2232..2250)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2255..2271)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2255..2271)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2373..2392)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2384..3193
                     /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(2451..2470)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3340..3928
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3829..3848
                     /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 accatgggac ctggaccacc tgcagcagga gcagccccaa gcccccgccc
      721 tctgtccctg gtggcaaggc tgagctacgc agtgggccac tttctgaacg acctgtgcgc
      781 ctccatgtgg ttcacctacc tgctgctgta tctgcactct gtgagagcat atagctccag
      841 gggagcagga ctgctgctgc tgctgggaca ggtggcagac ggactgtgca caccactggt
      901 gggatacgag gcagataggg cagcaagctg ctgtgcaaga tatggaccaa ggaaggcatg
      961 gcacctggtg ggaaccgtgt gcgtgctgct gtctttcccc tttatcttca gcccttgcct
     1021 gggctgtgga gcagcaacac cagagtgggc cgccctgctg tactatggcc cttttatcgt
     1081 gatctttcag ttcggctggg cctccaccca gatctcccac ctgtctctga tccccgagct
     1141 ggtgaccaac gaccacgaga aggtggagct gacagccctg agatacgcct tcaccgtggt
     1201 ggcaaatatc acagtgtatg gagcagcatg gctgctgctg cacctgcagg gatctagcag
     1261 ggtggagcca acccaggaca tcagcatctc cgatcagctg ggaggacagg atgtgcccgt
     1321 gttccggaac ctgagcctgc tggtggtggg agtgggagcc gtgttttccc tgctgttcca
     1381 cctgggcacc agagagcgga gaaggccaca cgccgaggag ccaggagagc acacccctct
     1441 gctggcacca gcaacagcac agccactgct gctgtggaag cactggctga gagagcccgc
     1501 cttttaccaa gtgggcatcc tgtatatgac cacaaggctg atcgtgaatc tgtctcagac
     1561 ctacatggcc atgtacctga catatagcct gcacctgcct aagaagttca tcgccaccat
     1621 cccactggtc atgtacctgt ctggctttct gtcctctttc ctgatgaagc ctatcaacaa
     1681 gtgcatcggc agaaatatga catacttcag cggactgctg gtcatcctgg ccttcgccgc
     1741 ctgggtggcc ctggccgagg gcctgggcgt ggccgtgtac gcagcagccg tgctgctggg
     1801 agcaggatgt gcaaccatcc tggtgacaag cctggcaatg accgcagacc tgatcggacc
     1861 acacacaaac tccggcgcct tcgtgtacgg ctctatgagc ttcctggata aggtggccaa
     1921 tggcctggcc gtgatggcca tccagtccct gcacccatgc ccctctgagc tgtgctgtag
     1981 ggcctgcgtg agcttctacc actgggccat ggtggcagtg accggaggag tgggagtggc
     2041 agccgccctg tgcctgtgct ctctgctgct gtggcctaca aggctgcgcc ggtgggacag
     2101 agatgcaagg ccagacccag ctttcttgta caaagttggc attataagaa agcattgctt
     2161 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     2221 agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
     2281 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac
     2341 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
     2401 ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     2461 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga
     2521 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     2581 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     2641 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca
     2701 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     2761 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     2821 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     2881 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
     2941 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     3001 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     3061 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     3121 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     3181 tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac
     3241 ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     3301 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3361 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3421 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3481 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3541 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3601 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3661 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3721 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3781 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3841 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     3901 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3961 ggccttttgc tggccttttg ctcacatgtt
//
