LOCUS       sequence-259219-        3987 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 3987)
  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 3987)
  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_RHCG
FEATURES             Location/Qualifiers
     source          1..3987
                     /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(2112..2211)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2228..2247)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2229..2247)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2252..2268)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2252..2268)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2370..2389)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2381..3190
                     /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(2448..2467)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3337..3925
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3826..3845
                     /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 accatggcct ggaacaccaa tctgaggtgg cgcctgccac tgacatgcct
      721 gctgctgcag gtcatcatgg tcatcctgtt cggcgtgttt gtgcggtacg acttcgaggc
      781 cgatgcccac tggtggtctg agagaaccca caagaacctg agcgacatgg agaatgagtt
      841 ctactatcgg taccccagct ttcaggatgt ccacgtaatg gtgtttgttg gtttcggctt
      901 tctgatgaca ttcctgcaga gatatggctt ttccgccgtg ggcttcaact ttctgctggc
      961 cgcctttggc atccagtggg ccctgctgat gcagggctgg ttccactttc tgcaggaccg
     1021 gtacatcgtg gtgggcgtgg agaacctgat caatgccgat ttctgcgtgg caagcgtgtg
     1081 cgtggccttt ggcgccgtgc tgggcaaggt gtccccaatc cagctgctga tcatgacctt
     1141 ctttcaggtg acactgttcg ccgtgaacga gtttatcctg ctgaatctgc tgaaggtgaa
     1201 ggacgccggc ggctctatga ccatccacac attcggcgcc tactttggcc tgaccgtgac
     1261 aaggatcctg tatcggagaa acctggagca gtccaaggag cgccagaata gcgtgtacca
     1321 gtccgacctg ttcgccatga tcggcaccct gtttctgtgg atgtactggc ctagcttcaa
     1381 ctccgccatc tcttatcacg gcgatagcca gcacagggca gccatcaata cctattgctc
     1441 cctggcagca tgcgtgctga ccagcgtggc catcagctcc gccctgcaca agaagggcaa
     1501 gctggacatg gtgcacatcc agaacgccac cctggcagga ggagtggcag tgggcacagc
     1561 cgccgagatg atgctgatgc catacggcgc cctgatcatc ggcttcgtgt gcggcatcat
     1621 ctctaccctg ggcttcgtgt atctgacacc ctttctggag agcagactgc acatccagga
     1681 tacctgtggc atcaacaatc tgcacggcat ccctggaatc atcggaggaa tcgtgggagc
     1741 agtgacagca gcatccgcct ctctggaggt gtacggcaag gagggcctgg tgcacagctt
     1801 cgactttcag ggcttcaatg gcgattggac cgccaggaca cagggcaagt ttcagatcta
     1861 tggactgctg gtgaccctgg caatggcact gatgggaggc atcatcgtgg gcctgatcct
     1921 gcgcctgcct ttctggggcc agccatccga cgagaactgt tttgaggatg ccgtgtactg
     1981 ggagatgcct gagggcaatt ctaccgtgta tatcccagag gatcccacat tcaagcctag
     2041 cggaccatcc gtgccatctg tgcctatggt gtcccccctg cctatggcct ctagcgtgcc
     2101 actggtgccc gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     2161 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     2221 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     2281 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     2341 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     2401 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     2461 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     2521 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     2581 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     2641 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     2701 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     2761 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     2821 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     2881 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     2941 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     3001 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     3061 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     3121 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     3181 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     3241 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     3301 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3361 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3421 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3481 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3541 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3601 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3661 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3721 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3781 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3841 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3901 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3961 cttttgctgg ccttttgctc acatgtt
//
