LOCUS       sequence-355838-        4089 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 4089)
  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 4089)
  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-SPNS3_STOP
FEATURES             Location/Qualifiers
     source          1..4089
                     /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(1646..1745)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1762..1781)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1763..1781)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1786..1802)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1786..1802)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1904..1923)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1915..2724
                     /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(1982..2001)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2871..3459
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3360..3379
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3613..3630
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3789..3816)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3908..3994)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4043..4065
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4057..4074
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4058..4074
                     /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 ccaccatggc aggaggaatg
      121 tctgccgagt gcccagagcc tggaccagga ggactgcagg gccagagccc cggccctggc
      181 cggcagtgtc cacctccaat cacccccaca agctggtccc tgccaccttg gagagcatac
      241 gtggcagcag ccgtgctgtg ctacatcaac ctgctgaatt atatgaactg gttcatcatc
      301 gccggcgtgc tgctggacat ccaggaggtg tttcagatct ccgataatca cgccggcctg
      361 ctgcagaccg tgttcgtgtc ctgtctgctg ctgtctgccc ccgtgttcgg ctatctgggc
      421 gacaggcact cccgcaaggc cacaatgtct ttcggcatcc tgctgtggtc cggagcagga
      481 ctgagctcct cttttatctc tccaaggtac agctggctgt tctttctgag caggggaatc
      541 gtgggaaccg gatctgccag ctattccacc atcgccccaa cagtgctggg cgacctgttc
      601 gtgagggatc agcggacaag agtgctggcc gtgttctaca tctttatccc agtgggaagc
      661 ggactgggat atgtgctggg atccgccgtg accatgctga caggcaactg gcggtgggcc
      721 ctgagagtga tgccttgcct ggaggccgtg gccctgatcc tgctgatcct gctggtgcca
      781 gacccaccca ggggagcagc agagacccag ggagagggag ccgtgggcgg ctttcggagc
      841 tcctggtgtg aggatgtgag atacctgggc aagaattggt ccttcgtgtg gtctaccctg
      901 ggcgtgacag ccatggcctt tgtgacaggc gccctgggat tctgggcacc aaagtttctg
      961 ctggaggcaa gggtggtgca cggactgcag cctccatgct tccaggagcc atgtagcaac
     1021 cccgactccc tgatctttgg cgccctgacc atcatgacag gcgtgatcgg cgtgatcctg
     1081 ggagcagagg cagcacggag atataagaaa gtgatccctg gagcagagcc actgatctgc
     1141 gcatctagcc tgctggcaac cgcaccatgt ctgtacctgg ccctggtgct ggcaccaacc
     1201 acactgctgg catcttacgt gttcctggga ctgggagagc tgctgctgag ctgcaattgg
     1261 gccgtggtgg ccgatatcct gctgtccgtg gtggtgccaa ggtgtagggg aaccgcagag
     1321 gccctgcaga tcacagtggg acacatcctg ggcgacgcag gatcccccta cctgaccggc
     1381 ctgatctcct ctgtgctgag ggcaaggcgc cctgatagct atctgcagcg gttcagatct
     1441 ctgcagcaga gcttcctgtg ctgtgccttt gtgatcgccc tgggaggagg atgctttctg
     1501 ctgaccgccc tgtacctgga gagggatgag acaagggcat ggcagccagt gaccggaaca
     1561 cctgacagca atgacgtgga ttccaacgat ctggagagac agggcctgct gtctggcgcc
     1621 ggcgccagca ccgaggagcc ttaagaccca gctttcttgt acaaagttgg cattataaga
     1681 aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt
     1741 atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc
     1801 tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata
     1861 tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc
     1921 catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat
     1981 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat
     2041 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
     2101 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
     2161 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa
     2221 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
     2281 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat
     2341 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
     2401 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa
     2461 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
     2521 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
     2581 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
     2641 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
     2701 ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc
     2761 attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac
     2821 gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     2881 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2941 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3001 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     3061 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3121 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3181 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3241 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3301 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3361 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3421 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3481 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3541 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     3601 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     3661 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     3721 aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga agagtttgta
     3781 gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt
     3841 atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc
     3901 ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg
     3961 cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgcg
     4021 ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaag
     4081 ctcgggccc
//
