LOCUS       sequence-259329-        3945 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 3945)
  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 3945)
  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_MFSD1
FEATURES             Location/Qualifiers
     source          1..3945
                     /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(2070..2169)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2186..2205)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2187..2205)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2210..2226)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2210..2226)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2328..2347)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2339..3148
                     /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(2406..2425)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3295..3883
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3784..3803
                     /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 accatggagg aggaggacga ggaggcacgg gccctgctgg caggaggacc
      721 cgacgaggcc gatagaggcg cccccgccgc ccctggcgcc ctgcctgccc tgtgcgatcc
      781 atctcggctg gcccacagac tgctggtgct gctgctgatg tgcttcctgg gctttggcag
      841 ctacttctgt tatgacaacc ctgccgccct gcagacccag gtgaagcggg atatgcaagt
      901 gaataccaca aagtttatgc tgctgtacgc ctggtattcc tggccaaacg tggtgctgtg
      961 cttcttcggc ggcttcctga tcgacagggt gtttggcatc cgctggggca caatcatctt
     1021 ctcttgcttc gtgtgcatcg gacaggtggt gttcgcactg ggaggcatct tcaacgcctt
     1081 ttggctgatg gagtttggcc ggttcgtgtt tggaatcgga ggagagagcc tggcagtggc
     1141 acagaacacc tacgccgtgt cctggttcaa gggcaaggag ctgaatctgg tgtttggcct
     1201 gcagctgtcc atggccagaa tcggctctac agtgaacatg aatctgatgg gctggctgta
     1261 tagcaagatc gaggccctgc tgggatccgc cggacacacc acactgggaa tcaccctgat
     1321 gatcggcggc atcacatgca tcctgagcct gatctgtgcc ctggccctgg cctacctgga
     1381 tcagagggcc gagcgcatcc tgcacaagga gcagggcaag accggcgaag tgatcaagct
     1441 gacagacgtg aaggatttct ccctgccact gtggctgatc tttatcatct gcgtgtgcta
     1501 ctatgtggcc gtgttcccct ttatcggcct gggcaaggtg ttctttaccg agaagttcgg
     1561 ctttagctcc caggccgcca gcgccatcaa ttccgtggtg tatgtgatct ctgccccaat
     1621 gagccccgtg ttcggcctgc tggtggacaa gacaggcaag aacatcatct gggtgctgtg
     1681 cgcagtggca gccaccctgg tgagccacat gatgctggcc tttacaatgt ggaatccttg
     1741 gatcgccatg tgcctgctgg gcctgtctta cagcctgctg gcatgtgcac tgtggccaat
     1801 ggtggccttc gtggtccctg agcaccagct gggaaccgcc tatggcttta tgcagtctat
     1861 ccagaacctg ggcctggcca tcatctctat catcgccggc atgatcctgg atagcagggg
     1921 ctacctgttc ctggaggtgt tctttatcgc ctgcgtgagc ctgtctctgc tgtccgtggt
     1981 gctgctgtac ctggtgaatc gcgcccaggg cggcaacctg aattattccg cccggcagag
     2041 agaggagatc aagttctctc acaccgagga cccagctttc ttgtacaaag ttggcattat
     2101 aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat
     2161 cattatttgc catccagctg atatccccta tagtgagtcg tattacatgg tcatagctgt
     2221 ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa
     2281 tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat
     2341 gagccatatt caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt
     2401 atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt
     2461 gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa
     2521 tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac
     2581 catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg
     2641 aaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc
     2701 gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag
     2761 cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc
     2821 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca
     2881 taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa
     2941 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc
     3001 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt
     3061 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt
     3121 tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggca
     3181 gagcattacg ctgacttgac gggacggcgc aagctcatga ccaaaatccc ttaacgtgag
     3241 ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3301 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3361 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3421 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     3481 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3541 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3601 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3661 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3721 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3781 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3841 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3901 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtt
//
