LOCUS       sequence-345622-        3474 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 3474)
  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 3474)
  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-UCP1_STOP
FEATURES             Location/Qualifiers
     source          1..3474
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1031..1130)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1147..1166)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1148..1166)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1171..1187)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1171..1187)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1289..1308)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1300..2109
                     /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(1367..1386)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2256..2844
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2745..2764
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2998..3015
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3174..3201)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3293..3379)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3428..3450
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3442..3459
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3443..3459
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacacat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccaccatggg aggactgacc
      121 gcatctgacg tgcaccccac actgggagtg cagctgttca gcgccggaat cgcagcatgc
      181 ctggcagacg tgatcacctt tcctctggat acagccaagg tgaggctgca ggtgcagggc
      241 gagtgtccaa ccagctccgt gatccgctac aagggcgtgc tgggcaccat cacagccgtg
      301 gtgaagacag agggccggat gaagctgtat tccggactgc ctgcaggact gcagaggcag
      361 atctctagcg cctctctgcg catcggcctg tacgataccg tgcaggagtt cctgaccgcc
      421 ggcaaggaga cagccccatc tctgggcagc aagatcctgg caggactgac cacaggagga
      481 gtggccgtgt ttatcggaca gccaaccgag gtggtgaagg tgaggctgca ggcacagagc
      541 cacctgcacg gcatcaagcc tcggtacacc ggcacataca acgcctatag aatcatcgca
      601 accacagagg gactgaccgg actgtggaag ggaaccacac ccaacctgat gcgcagcgtg
      661 atcatcaatt gcaccgagct ggtgacatat gacctgatga aggaggcctt cgtgaagaac
      721 aatatcctgg ccgacgatgt gccatgccac ctggtgtccg ccctgatcgc aggattctgt
      781 gcaaccgcca tgtcctctcc cgtggatgtg gtgaagacac ggtttatcaa cagcccccct
      841 ggccagtata agtccgtgcc taactgtgcc atgaaggtgt tcaccaatga gggcccaaca
      901 gccttcttta agggcctggt gccctccttt ctgagactgg gctcttggaa tgtgatcatg
      961 ttcgtgtgct ttgagcagct gaagcgggag ctgagcaagt ccagacagac catggactgt
     1021 gccacataag acccagcttt cttgtacaaa gttggcatta taagaaagca ttgcttatca
     1081 atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg ccatccagct
     1141 gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc
     1201 gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata
     1261 aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa
     1321 acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     1381 cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg
     1441 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     1501 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
     1561 actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta
     1621 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     1681 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc
     1741 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     1801 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
     1861 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     1921 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     1981 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     2041 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     2101 tttttctaat cagaattggt taattggttg taacactggc agagcattac gctgacttga
     2161 cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc gttccactga
     2221 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     2281 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     2341 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     2401 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     2461 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     2521 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     2581 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     2641 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     2701 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     2761 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     2821 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     2881 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     2941 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     3001 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     3061 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     3121 cgcaacgcaa ttaatacgcg taccgctagc caggaagagt ttgtagaaac gcaaaaaggc
     3181 catccgtcag gatggccttc tgcttagttt gatgcctggc agtttatggc gggcgtcctg
     3241 cccgccaccc tccgggccgt tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc
     3301 tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag tcttccgact
     3361 gagcctttcg ttttatttga tgcctggcag ttccctactc tcgcgttaac gctagcatgg
     3421 atgttttccc agtcacgacg ttgtaaaacg acggccagtc ttaagctcgg gccc
//
