LOCUS       sequence-263226-        3477 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 3477)
  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 3477)
  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_UCP2
FEATURES             Location/Qualifiers
     source          1..3477
                     /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(1602..1701)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1718..1737)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1719..1737)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1742..1758)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1742..1758)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1860..1879)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1871..2680
                     /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(1938..1957)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2827..3415
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3316..3335
                     /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 accatggtgg gcttcaaggc aaccgacgtg ccacctaccg caacagtgaa
      721 gtttctggga gcaggaacag cagcatgcat cgcagacctg atcaccttcc ctctggatac
      781 agccaaggtg aggctgcaga tccagggcga gtctcaggga ccagtgcggg caaccgcaag
      841 cgcccagtac agaggcgtga tgggcaccat cctgacaatg gtgaggacag agggccctcg
      901 ctccctgtat aatggactgg tggcaggact gcagaggcag atgtctttcg ccagcgtgcg
      961 catcggcctg tacgactccg tgaagcagtt ttataccaag ggctctgagc acgcctccat
     1021 cggctctaga ctgctggcag gatctaccac aggcgccctg gcagtggcag tggcacagcc
     1081 aaccgatgtg gtgaaggtga ggttccaggc acaggcaagg gcaggaggag gccggagata
     1141 ccagtctacc gtgaacgcct ataagacaat cgcccgggag gagggcttta gaggcctgtg
     1201 gaagggcacc agcccaaacg tggcaaggaa tgccatcgtg aactgtgccg agctggtgac
     1261 atacgacctg atcaaggatg ccctgctgaa ggccaatctg atgaccgacg atctgccctg
     1321 ccacttcaca agcgcctttg gcgccggctt ctgtaccaca gtgatcgcca gccctgtgga
     1381 tgtggtgaag accagataca tgaactccgc cctgggacag tatagctccg ccggacactg
     1441 cgcactgaca atgctgcaga aggagggccc aagggccttt tacaagggct tcatgcccag
     1501 ctttctgcgc ctgggctcct ggaacgtggt catgttcgtg acctatgagc agctgaagcg
     1561 ggccctgatg gcagcatgta catccagaga ggcccccttt gacccagctt tcttgtacaa
     1621 agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag
     1681 tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat
     1741 ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc
     1801 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
     1861 aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat
     1921 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     1981 aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     2041 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     2101 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     2161 tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     2221 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     2281 tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
     2341 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     2401 gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt
     2461 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     2521 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     2581 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     2641 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     2701 gtaacactgg cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc
     2761 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2821 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     2881 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     2941 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3001 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3061 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3121 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3181 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3241 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3301 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3361 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3421 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
