LOCUS       sequence-312076-        3789 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 3789)
  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 3789)
  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-MFSD3_STOP
FEATURES             Location/Qualifiers
     source          1..3789
                     /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(1914..2013)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2030..2049)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2031..2049)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2054..2070)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2054..2070)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2172..2191)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2183..2992
                     /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(2250..2269)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3139..3727
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3628..3647
                     /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 accatgaggg gcaagctgct gcctctggca ggactgtacc tggtgcaggg
      721 actgccatat ggactgcagt ccggactgct gcctgtgctg ctgagggcag gcggcctgtc
      781 tctgaccaga gtgggcctgg ccaaggtgct gtacgcacca tggctgctga agctggcatg
      841 ggcaccactg gtggacgcac agggcagcgc cagggcatgg gtgaccagat ccacagcagg
      901 actgggactg gtgtgcggcc tgctggccgg cctgccccct ccaggagcag gacaggcagg
      961 actgccagca gcagtggcag gactgctgct gctgctgaac ctgggagcag caatgcagga
     1021 cgtggccctg gatgcactgg cagtgcagct gctggagcct gcagagctgg gcccaggcaa
     1081 taccgtgcag gtggtggcct acaagctggg agccgccctg gcaggaggcg ccctgctggc
     1141 cctgctgccc accttcagct ggcctcagct gtttctgctg ctggcagcaa catattggct
     1201 ggcagccgcc ctggcatggg cagcaccagc cctgcggaga ctgccacagc agccaccttc
     1261 cgagcagagg ccacacaccg cacacctgct gagggacgtg ctggcagtgc ctggaaccgt
     1321 gtggacagca ggcttcgtgc tgacatataa gctgggagag cagggagcaa gctccctgtt
     1381 tcctctgctg ctgctggatc acggcgtgtc tgccccagag ctgggactgt ggaacggagt
     1441 gggagcagtg gtgtgcagca tcgcaggatc tagcctggga ggcaccctgc tggcaaagca
     1501 ctggaagctg ctgccactgc tgcggtccgt gctgaggttc cgcctgggag gactggcatg
     1561 tcagaccgcc ctggtgtttc acctggacac actgggcgcc tctatggatg ccggcacaat
     1621 cctgagaggc tccgccctgc tgtctctgtg cctgcagcac ttcctgggcg gcctggtgac
     1681 cacagtgacc tttacaggca tgatgaggtg tagccagctg gcaccacgcg ccctgcaggc
     1741 aacccactac tccctgctgg ccacactgga gctgctgggc aagctgctgc tgggcaccct
     1801 ggcaggagga ctggcagacg gactgggacc acacccctgt ttcctgctgc tgctgatcct
     1861 gtctgccttt cctgtgctgt atctggatct ggccccaagc acattcctgt aagacccagc
     1921 tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca
     1981 ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga
     2041 gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg
     2101 atgttacatt gcacaagata aaataatatc atcatgaaca ataaaactgt ctgcttacat
     2161 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt
     2221 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     2281 atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa
     2341 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     2401 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     2461 gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga
     2521 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     2581 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     2641 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     2701 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac
     2761 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     2821 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     2881 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     2941 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     3001 ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc
     3061 atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga
     3121 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3181 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3241 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     3301 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3361 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3421 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3481 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3541 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3601 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3661 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3721 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     3781 tcacatgtt
//
