LOCUS       sequence-213592-        2860 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Plasmid for the insect cell expression system, donor for plasmid 
            fusions, two promoters, Multiplication Module M, LoxP site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2860)
  AUTHORS   Fitzgerald DJ, Berger P, Schaffitzel C, Yamada K, Richmond TJ, 
            Berger I
  TITLE     Protein complex expression by using multigene baculoviral vectors.
  JOURNAL   Nat Methods. 2006 Dec;3(12):1021-32. doi: 10.1038/nmeth983.
  PUBMED    17117155
REFERENCE   2  (bases 1 to 2860)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName: 
            "Nat Methods"; date: "2006-12"; volume: "3"; issue: "12"; pages: 
            "1021-32"
COMMENT     Sequence Label: pSPL
FEATURES             Location/Qualifiers
     source          1..2860
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      303..659
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    complement(712..745)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    935..983
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     1008..1027
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(1119..1228)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        1273..1364
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     1289..1306
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     polyA_signal    1623..1757
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1660..1679)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1714..1733
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(1907..2698)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg agcgcatacg
       61 ctacttaccg gtttattgac taccggaagc agtgtgaccg tgtgcttctc aaatgcctga
      121 ggccagtttg ctcaggctct ccccgtggag gtaataattg acgatatgat catttattct
      181 gcctcccagc tgacattcat ccggggtcag caccgtttct gcggactggc tttctacgtg
      241 ttccgcttcc tttagcagcc cttgcgccct gagtgcttgc ggcagcgtga agctaattcc
      301 catgtcagcc gttaagtgtt cctgtgtcac tcaaaattgc tttgagaggc tctaagggct
      361 tctcagtgcg ttacatccct ggcttgttgt ccacaaccgt taaaccttaa aagctttaaa
      421 agccttatat attctttttt ttcttataaa acttaaaacc ttagaggcta tttaagttgc
      481 tgatttatat taattttatt gttcaaacat gagagcttag tacgtgaaac atgagagctt
      541 agtacgttag ccatgagagc ttagtacgtt agccatgagg gtttagttcg ttaaacatga
      601 gagcttagta cgttaaacat gagagcttag tacgtgaaac atgagagctt agtacgtact
      661 atcaacaggt tgaactgctg atcaacagat cctctacgcg gccgcggtac cataacttcg
      721 tatagcatac attatacgaa gttatctggt ttaaacgtac ccgtagtggc tatggcaggg
      781 cttgccgccc cgacgttggc tgcgagccct gggccttcac ccgaacttgg gggttggggt
      841 ggggaaaagg aagaaacgcg ggcgtattgg tcccaatggg gtctcggtgg ggtatcgaca
      901 gagtgccagc cctgggaccg aaccccgcgt ttatgaacaa acgacccaac acccgtgcgt
      961 tttattctgt ctttttattg ccgtcatagc gcgggttcct tccggtattg tctccttccg
     1021 tgtttcagtt agcctccccc atctcccggt accgcatgct atgcatcagc tgctagcacc
     1081 atggctcgag atcccgggtg atcaagtctt cgtcgagtga ttgtaaataa aatgtaattt
     1141 acagtatagt attttaatta atatacaaat gatttgataa taattcttat ttaactataa
     1201 tatattgtgt tgggttgaat taaaggtccg tatactagta tcgattcgcg acctactccg
     1261 gaatattaat agatcatgga gataattaaa atgataacca tctcgcaaat aaataagtat
     1321 tttactgttt tcgtaacagt tttgtaataa aaaaacctat aaatattccg gattattcat
     1381 accgtcccac catcgggcgc ggatcccggt ccgaagcgcg cggaattcaa aggcctacgt
     1441 cgacgagctc actagtcgcg gccgctttcg aatctagagc ctgcagtctc gacaagcttg
     1501 tcgagaagta ctagaggatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     1561 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     1621 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     1681 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     1741 cttatcatgt ctggatctga tcactgcttg agcctagaag atccggctgc taacaaagcc
     1801 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactatcata acccctagga
     1861 gtccaagcga gctcgatatc cgtcggcttg aacgaattgt tagacattat ttgccgacta
     1921 ccttggtgat ctcgcctttc acgtagtgga caaattcttc caactgatct gcgcgcgagg
     1981 ccaagcgatc ttcttcttgt ccaagataag cctgtctagc ttcaagtatg acgggctgat
     2041 actgggccgg caggcgctcc attgcccagt cggcagcgac atccttcggc gcgattttgc
     2101 cggttactgc gctgtaccaa atgcgggaca acgtaagcac tacatttcgc tcatcgccag
     2161 cccagtcggg cggcgagttc catagcgtta aggtttcatt tagcgcctca aatagatcct
     2221 gttcaggaac cggatcaaag agttcctccg ccgctggacc taccaaggca acgctatgtt
     2281 ctcttgcttt tgtcagcaag atagccagat caatgtcgat cgtggctggc tcgaagatac
     2341 ctgcaagaat gtcattgcgc tgccattctc caaattgcag ttcgcgctta gctggataac
     2401 gccacggaat gatgtcgtcg tgcacaacaa tggtgacttc tacagcgcgg agaatctcgc
     2461 tctctccagg ggaagccgaa gtttccaaaa ggtcgttgat caaagctcgc cgcgttgttt
     2521 catcaagcct tacggtcacc gtaaccagca aatcaatatc actgtgtggc ttcaggccgc
     2581 catccactgc ggagccgtac aaatgtacgg ccagcaacgt cggttcgaga tggcgctcga
     2641 tgacgccaac tacctctgat agttgagtcg atacttcggc gatcaccgct tccctcatga
     2701 tgtttaactt tgttttaggg cgactgccct gctgcgtaac atcgttgctg ctccataaca
     2761 tcaaacatcg acccacggcg taacgcgctt gctgcttgga tgcccgaggc atagactgta
     2821 ccccaaaaaa acagtcataa caagccatga aaaccgccac
//
