LOCUS       sequence-259098-        4179 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 4179)
  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 4179)
  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_MFSD2A
FEATURES             Location/Qualifiers
     source          1..4179
                     /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(2304..2403)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2420..2439)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2421..2439)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2444..2460)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2444..2460)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2562..2581)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2573..3382
                     /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(2640..2659)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3529..4117
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4018..4037
                     /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 accatggcca agggagaggg agcagagagc ggatccgccg ccggcctgct
      721 gccaaccagc atcctgcagt ccacagagcg ccctgcccag gtgaagaagg agcccaagaa
      781 gaagaagcag cagctgtccg tgtgcaacaa gctgtgctac gcactgggag gagcacctta
      841 tcaggtgacc ggatgcgccc tgggcttctt tctgcagatc tacctgctgg acgtggccca
      901 gaaggatgag gaggtggtgt tctgttttag ctcctttcaa gtgggcccat tctctgccag
      961 catcatcctg tttgtgggaa gggcatggga cgcaatcacc gatccactgg tgggactgtg
     1021 catctccaag tctccttgga catgtctggg ccggctgatg ccctggatca tcttcagcac
     1081 ccctctggcc gtgatcgcct acttcctgat ctggtttgtg cccgacttcc ctcacggcca
     1141 gacatactgg tatctgctgt tttactgcct gttcgagacc atggtgacat gttttcacgt
     1201 gccctattcc gccctgacca tgttcatctc taccgagcag acagagcgcg attctgccac
     1261 cgcctatcgg atgacagtgg aggtgctggg aaccgtgctg ggaacagcaa tccagggaca
     1321 gatcgtggga caggcagaca ccccttgctt ccaggatctg aactctagca ccgtggccag
     1381 ccagtccgcc aatcacacac acggcaccac aagccaccgg gagacccaga aggcatacct
     1441 gctggcagca ggcgtgatcg tgtgcatcta tatcatctgt gccgtgatcc tgatcctggg
     1501 cgtgcgcgag cagagggagc cttacgaggc ccagcagtct gagccaatcg cctacttccg
     1561 gggactgagg ctggtcatga gccacggccc ctacatcaag ctgatcaccg gcttcctgtt
     1621 tacaagcctg gccttcatgc tggtggaggg caacttcgtg ctgttttgta cctatacact
     1681 gggctttaga aacgagttcc agaatctgct gctggccatc atgctgtccg ccaccctgac
     1741 aatcccaatc tggcagtggt ttctgaccag gttcggcaag aagacagccg tgtacgtggg
     1801 catctcctct gccgtgccct ttctgatcct ggtggccctg atggagtcca atctgatcat
     1861 cacctatgcc gtggcagtgg cagcaggaat cagcgtggca gcagccttcc tgctgccatg
     1921 gtccatgctg cccgacgtga tcgacgattt tcacctgaag cagccacact tccacggcac
     1981 agagcccatc ttcttttcct tttacgtgtt ctttaccaag ttcgcctctg gcgtgagcct
     2041 gggcatctcc acactgtctc tggattttgc cggctatcag accagaggct gctctcagcc
     2101 agagagggtg aagttcaccc tgaacatgct ggtgacaatg gcccccatcg tgctgatcct
     2161 gctgggcctg ctgctgttca agatgtaccc tatcgacgag gagcggagaa ggcagaataa
     2221 gaaggccctg caggccctga gggacgaggc aagctcctct ggctgttctg agaccgatag
     2281 cacagagctg gccagcatcc tggacccagc tttcttgtac aaagttggca ttataagaaa
     2341 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat
     2401 ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg
     2461 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     2521 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     2581 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     2641 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     2701 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     2761 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     2821 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac
     2881 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     2941 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     3001 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     3061 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     3121 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     3181 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     3241 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     3301 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     3361 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat
     3421 tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc
     3481 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3541 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3601 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3661 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3721 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3781 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3841 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3901 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3961 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4021 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4081 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4141 acggttcctg gccttttgct ggccttttgc tcacatgtt
//
