LOCUS       sequence-253303-        4101 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 4101)
  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 4101)
  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_MFSD13A
FEATURES             Location/Qualifiers
     source          1..4101
                     /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(2226..2325)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2342..2361)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2343..2361)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2366..2382)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2366..2382)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2484..2503)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2495..3304
                     /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(2562..2581)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3451..4039
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3940..3959
                     /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 tgggacagcc tcaggcatgg ctgctgggac tgccaaccgc
      721 agtggtgtac ggaagcctgg ccctgtttac cacaatcctg cacaacgtgt tcctgctgta
      781 ctatgtggat accttcgtga gcgtgtacaa gatcaataag atggcctttt gggtgggcga
      841 gacagtgttc ctgctgtgga acagcctgaa tgacccactg ttcggctggc tgtccgatag
      901 acagtttctg agctcccagc ccaggagcgg agcaggactg tctagcaggg cagtggtgct
      961 ggcccgggtg caggccctgg gctggcacgg accactgctg gccctgtcct tcctggcctt
     1021 ttgggtgcca tgggcacctg caggactgca gttcctgctg tgcctgtgcc tgtacgacgg
     1081 ctttctgacc ctggtggatc tgcaccacca cgcactgctg gcagacctgg ccctgtccgc
     1141 ccacgatcgg acacacctga acttttactg cagcctgttc tccgccgcag gatctctgag
     1201 cgtgtttgcc tcttatgcct tctggaataa ggaggacttc tcctctttta gagccttctg
     1261 cgtgaccctg gccgtgagct ccggactggg cttcctggga gcaacacagc tgctgcggag
     1321 aagggtggag gcagcaagga aggaccctgg atgctccgga ctggtggtgg attctggact
     1381 gtgcggagag gagctgctgg tgggatctga ggaggccgac agcatcaccc tgggcagata
     1441 cctgaggcag ctggcccgcc accggaactt tctgtggttc gtgagcatgg atctggtgca
     1501 ggtgtttcac tgccacttca actctaattt ctttcctctg ttcctggagc acctgctgag
     1561 cgaccacatc tccctgtcta caggcagcat cctgctgggc ctgtcctacg tggccccaca
     1621 cctgaacaat ctgtatttcc tgtctctgtg ccgccggtgg ggcgtgtatg cagtggtgag
     1681 aggcctgttt ctgctgaagc tgggactgag cctgctgatg ctgctggcag gaccagatca
     1741 cctgtccctg ctgtgcctgt ttatcgcctc taaccgcgtg ttcaccgagg gcacctgtaa
     1801 gctgctgacc ctggtggtga cagacctggt ggacgaggat ctggtgctga atcacaggaa
     1861 gcaggcagca tccgccctgc tgttcggaat ggtggccctg gtgaccaagc caggccagac
     1921 atttgcccca ctgctgggaa cctggctgct gtgcttttac acaggccacg atctgttcca
     1981 gcagtctctg atcacccctg tgggcagcgc ccacccatgg ccagagccac ctgcacctgc
     2041 accagcacag gcccccaccc tgcgccaggg ctgcttctat ctgctggtgc tggtgcctat
     2101 cacatgtgcc ctgctgcagc tgtttacctg gtcccagttc acactgcacg gcagaaggct
     2161 gcacatggtg aaggcccagc ggcagaatct gtctcaggcc cagaccctgg acgtgaagat
     2221 ggtggaccca gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     2281 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     2341 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     2401 tcaaaatctc tgatgttaca ttgcacaaga taaaataata tcatcatgaa caataaaact
     2461 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     2521 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     2581 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     2641 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     2701 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     2761 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     2821 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     2881 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     2941 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     3001 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     3061 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     3121 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     3181 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     3241 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     3301 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     3361 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     3421 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3481 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3541 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     3601 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3661 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3721 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3781 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3841 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3901 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3961 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4021 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4081 ctggcctttt gctcacatgt t
//
