LOCUS       sequence-258993-        3777 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 3777)
  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 3777)
  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_RHAG
FEATURES             Location/Qualifiers
     source          1..3777
                     /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(1902..2001)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2018..2037)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2019..2037)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2042..2058)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2042..2058)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2160..2179)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2171..2980
                     /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(2238..2257)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3127..3715
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3616..3635
                     /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 accatgcggt tcacctttcc tctgatggcc atcgtgctgg agatcgccat
      721 gatcgtgctg ttcggcctgt ttgtggagta cgagaccgac cagacagtgc tggagcagct
      781 gaacatcacc aagcctacag acatgggcat cttctttgag ctgtacccac tgtttcagga
      841 tgtgcacgtg atgatcttcg tgggcttcgg ctttctgatg acctttctga agaagtatgg
      901 cttcagctcc gtgggcatca acctgctggt ggccgccctg ggactgcagt ggggcacaat
      961 cgtgcagggc atcctgcaga gccagggcca gaagtttaat atcggcatca agaacatgat
     1021 caatgccgac ttttccgccg ccaccgtgct gatctctttc ggcgccgtgc tgggcaagac
     1081 ctctccaaca cagatgctga tcatgacaat cctggagatc gtgttctttg cccacaacga
     1141 gtatctggtg agcgagatct tcaaggccag cgatatcggc gcctccatga ccatccacgc
     1201 cttcggcgca tactttggac tggcagtggc aggaatcctg tatcggtccg gcctgagaaa
     1261 gggccacgag aatgaggagt ccgcctacta ttctgacctg tttgccatga tcggcacact
     1321 gttcctgtgg atgttttggc caagcttcaa ctccgccatc gcagagccag gcgataagca
     1381 gtgcagggcc atcgtgaata cctacttctc tctggcagca tgcgtgctga cagccttcgc
     1441 cttttctagc ctggtggagc accgcggcaa gctgaacatg gtgcacatcc agaatgccac
     1501 cctggcagga ggagtggcag tgggaacatg cgccgacatg gccatccacc cattcggctc
     1561 tatgatcatc ggcagcatcg ccggcatggt gtccgtgctg ggctataagt ttctgacccc
     1621 cctgttcacc acaaagctgc ggatccacga tacatgtgga gtgcacaacc tgcacggact
     1681 gcctggagtg gtgggaggac tggccggcat cgtggccgtg gccatgggcg cctctaatac
     1741 cagcatggca atgcaggcag ccgccctggg atcctctatc ggaaccgcag tggtgggagg
     1801 actgatgaca ggcctgatcc tgaagctgcc cctgtggggc cagcctagcg atcagaattg
     1861 ctacgacgat tccgtgtatt ggaaggtgcc caagaccagg gacccagctt tcttgtacaa
     1921 agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag
     1981 tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat
     2041 ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc
     2101 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
     2161 aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat
     2221 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     2281 aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     2341 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     2401 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     2461 tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     2521 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     2581 tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
     2641 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     2701 gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt
     2761 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     2821 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     2881 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     2941 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     3001 gtaacactgg cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc
     3061 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3121 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3181 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3241 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3301 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3361 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3421 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3481 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3541 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3601 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3661 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3721 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
