LOCUS       sequence-312008-        3780 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 3780)
  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 3780)
  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_STOP
FEATURES             Location/Qualifiers
     source          1..3780
                     /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(1905..2004)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2021..2040)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2022..2040)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2045..2061)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2045..2061)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2163..2182)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2174..2983
                     /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(2241..2260)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3130..3718
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3619..3638
                     /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 taagacccag ctttcttgta
     1921 caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat
     1981 cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta
     2041 catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat
     2101 tgcacaagat aaaataatat catcatgaac aataaaactg tctgcttaca taaacagtaa
     2161 tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa
     2221 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     2281 gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     2341 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     2401 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     2461 cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     2521 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     2581 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     2641 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     2701 ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     2761 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     2821 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     2881 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     2941 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg
     3001 gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa
     3061 atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3121 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     3181 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     3241 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     3301 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     3361 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     3421 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     3481 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     3541 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     3601 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     3661 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     3721 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
//
