LOCUS       sequence-263070-        3768 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 3768)
  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 3768)
  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-NIPAL3
FEATURES             Location/Qualifiers
     source          1..3768
                     /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(1893..1992)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2009..2028)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2010..2028)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2033..2049)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2033..2049)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2151..2170)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2162..2971
                     /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(2229..2248)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3118..3706
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3607..3626
                     /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 accatggacg gaagccactc cgccgccctg aagctgcagc agctgccacc
      721 taccagctcc tctagcgccg tgtccgaggc ctctttcagc tacaaggaga acctgatcgg
      781 cgccctgctg gccatctttg gccacctggt ggtgtccatc gccctgaatc tgcagaagta
      841 ctgccacatc cggctggccg gctctaagga cccaagagcc tatttcaaga ccaagacatg
      901 gtggctgggc ctgtttctga tgctgctggg agagctgggc gtgttcgcaa gctatgcctt
      961 tgcccccctg tccctgatcg tgcctctgag cgccgtgtcc gtgatcgcct ctgccatcat
     1021 cggcatcatc ttcatcaagg agaagtggaa gcccaaggat ttcctgcgga gatacgtgct
     1081 gagctttgtg ggatgcggac tggcagtggt gggaacctat ctgctggtga cattcgcccc
     1141 taactctcac gagaagatga ccggcgagaa tgtgacacgg cacctggtga gctggccatt
     1201 tctgctgtac atgctggtgg agatcatcct gttctgtctg ctgctgtact tttataagga
     1261 gaagaacgct aacaatatcg tggtcatcct gctgctggtg gccctgctgg gcagcatgac
     1321 cgtggtgaca gtgaaggccg tggccggcat gctggtgctg tccatccagg gcaatctgca
     1381 gctggactac cctatcttct atgtgatgtt cgtgtgcatg gtggcaaccg ccgtgtacca
     1441 ggcagccttc ctgtctcagg ccagccagat gtacgattcc tctctgatcg cctccgtggg
     1501 ctatatcctg tctaccacaa tcgccatcac agccggcgcc atcttctatc tggactttat
     1561 cggcgaggat gtgctgcaca tctgcatgtt cgccctgggc tgtctgatcg ccttcctggg
     1621 cgtgtttctg atcaccagga accgcaagaa gccaatcccc tttgagccat acatctccat
     1681 ggacgccatg cccggcatgc agaatatgca cgataagggc atgacagtgc agcccgagct
     1741 gaaggcctcc ttttcttacg gcgccctgga gaacaatgat aacatcagcg agatctatgc
     1801 ccctgccacc ctgccagtga tgcaggagga gcacggaagc aggtccgcct ctggagtgcc
     1861 ttatcgcgtg ctggagcaca caaagaagga ggacccagct ttcttgtaca aagttggcat
     1921 tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa
     1981 aatcattatt tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc
     2041 tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     2101 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
     2161 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
     2221 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     2281 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     2341 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     2401 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     2461 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     2521 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     2581 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     2641 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     2701 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     2761 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     2821 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     2881 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     2941 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg
     3001 gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt
     3061 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3121 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3181 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3241 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     3301 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3361 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3421 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3481 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3541 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3601 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3661 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3721 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtt
//
