LOCUS       sequence-437194-        3531 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 3531)
  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 3531)
  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-Embigin
FEATURES             Location/Qualifiers
     source          1..3531
                     /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(1656..1755)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1772..1791)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1773..1791)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1796..1812)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1796..1812)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1914..1933)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1925..2734
                     /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(1992..2011)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2881..3469
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3370..3389
                     /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 ctctgcctgg actgctggaa gctcgggcta gaaccccacg
      721 actgctgctg ctgcagtgcc tgctggccgc cgcaagacct agctccgccg acggatccgc
      781 cccagattct cccttcacca gcccccctct gagggaggag atcatggcca acaatttttc
      841 tctggagagc cacaatatct ccctgacaga gcactctagc atgccagtgg agaagaacat
      901 caccctggag cgcccctcca acgtgaatct gacatgccag ttcaccacat ctggcgacct
      961 gaacgccgtg aatgtgacct ggaagaagga tggcgagcag ctggagaaca attacctggt
     1021 gtccgccacc ggctctacac tgtacaccca gtatcggttt accatcatca atagcaagca
     1081 gatgggcagc tattcctgct tctttcggga ggagaaggag cagagaggca ccttcaactt
     1141 caaggtgccc gagctgcacg gcaagaacaa gcctctgatc agctacgtgg gcgactccac
     1201 agtgctgacc tgcaagtgtc agaactgttt ccctctgaat tggacctggt attcctctaa
     1261 cggctctgtg aaggtgcccg tgggcgtgca gatgaacaag tacgtgatca atggcacata
     1321 tgccaacgag accaagctga agatcacaca gctgctggag gaggatggcg agagctactg
     1381 gtgcagggcc ctgttccagc tgggcgagtc tgaggagcac atcgagctgg tggtgctgag
     1441 ctatctggtg cctctgaagc ccttcctggt catcgtggcc gaagtgatcc tgctggtggc
     1501 caccatcctg ctgtgcgaga agtacacaca gaagaagaag aagcacagcg acgagggcaa
     1561 ggagtttgag cagatcgagc agctgaagtc cgatgatagc aacgggattg agaataatgt
     1621 gccacgccat agaaagaatg agtcactggg acaggaccca gctttcttgt acaaagttgg
     1681 cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa
     1741 taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat
     1801 agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga
     1861 taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg
     1921 tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc
     1981 tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta
     2041 tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt
     2101 tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct
     2161 tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat
     2221 ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt
     2281 tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt
     2341 taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt
     2401 tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga
     2461 aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact
     2521 tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg
     2581 aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc
     2641 ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt
     2701 gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca
     2761 ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa
     2821 cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     2881 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     2941 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3001 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     3061 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3121 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3181 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3241 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     3301 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     3361 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3421 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3481 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt t
//
