LOCUS       sequence-213581-        4943 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for the insect cell expression system, acceptor for plasmid 
            fusions, two promoters, Multiplication Module M, Tn7 transposition 
            and LoxP site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4943)
  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 4943)
  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 Methods"; date: "2006-12"; volume: "3"; issue: "12"; pages: 
            "1021-32"
COMMENT     Sequence Label: pKL
FEATURES             Location/Qualifiers
     source          1..4943
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    43..177
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(80..99)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     134..153
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  287..452
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     primer_bind     complement(723..742)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(808..1623)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(808..1623)
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1531..1550
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1615..1634
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1745..2333
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2234..2253
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2487..2504
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2638..2862
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2929..3462)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3651..3656,3657..3673,3674..3679))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(4075..4108)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    4298..4346
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4371..4390
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(4482..4591)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        4636..4727
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     4652..4669
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
ORIGIN
        1 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
       61 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
      121 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatctga
      181 tcactgcttg agcctagaag atccggctgc taacaaagcc cgaaaggaag ctgagttggc
      241 tgctgccacc gctgagcaat aactatcata acccctagga gatccgaacc agataagtga
      301 aatctagttc caaactattt tgtcattttt aattttcgta ttagcttacg acgctacacc
      361 cagttcccat ctattttgtc actcttccct aaataatcct taaaaactcc atttccaccc
      421 ctcccagttc ccaactattt tgtccgccca cagcggggca tttttcttcc tgttatgttt
      481 ttaatcaaac atcctgccaa ctccatgtga caaaccgtca tcttcggcta ctttttctct
      541 gtcacagaat gaaaattttt ctgtcatctc ttcgttatta atgtttgtaa ttgactgaat
      601 atcaacgctt atttgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
      661 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
      721 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccaaacgta ctgcggtaaa
      781 gctcatcagc gtggtcgtga agaattctta gaaaaactca tcgagcatca aatgaaactg
      841 caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga
      901 aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat
      961 tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc
     1021 aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat
     1081 ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc
     1141 aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt
     1201 aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc
     1261 aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg
     1321 gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg
     1381 aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc
     1441 aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg
     1501 atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc
     1561 agcatccatg ttggaattta atcgcggcct agagcaagac gtttcccgtt gaatatggct
     1621 cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc atgaccaaaa
     1681 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     1741 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     1801 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     1861 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     1921 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     1981 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     2041 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     2101 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     2161 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     2221 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     2281 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     2341 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     2401 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     2461 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     2521 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata gaccagccgc
     2581 gtaacctggc aaaatcggtt acggttgagt aataaatgga tgccctgcgt aagcgggtgt
     2641 gggcggacaa taaagtctta aactgaacaa aatagatcta aactatgaca ataaagtctt
     2701 aaactagaca gaatagttgt aaactgaaat cagtccagtt atgctgtgaa aaagcatact
     2761 ggacttttgt tatggctaaa gcaaactctt cattttctga agtgcaaatt gcccgtcgta
     2821 ttaaagaggg gcgtggccaa gggcatggta aagactatat tcgcggcgtt gtgacaattt
     2881 accgaacaac tccgcggccg ggaagccgat ctcggcttga acgaattgtt aggtggcggt
     2941 acttgggtcg atatcaaagt gcatcacttc ttcccgtatg cccaactttg tatagagagc
     3001 cactgcggga tcgtcaccgt aatctgcttg cacgtagatc acataagcac caagcgcgtt
     3061 ggcctcatgc ttgaggagat tgatgagcgc ggtggcaatg ccctgcctcc ggtgctcgcc
     3121 ggagactgcg agatcataga tatagatctc actacgcggc tgctcaaact tgggcagaac
     3181 gtaagccgcg agagcgccaa caaccgcttc ttggtcgaag gcagcaagcg cgatgaatgt
     3241 cttactacgg agcaagttcc cgaggtaatc ggagtccggc tgatgttggg agtaggtggc
     3301 tacgtctccg aactcacgac cgaaaagatc aagagcagcc cgcatggatt tgacttggtc
     3361 agggccgagc ctacatgtgc gaatgatgcc catacttgag ccacctaact ttgttttagg
     3421 gcgactgccc tgctgcgtaa catcgttgct gctgcgtaac atcgttgctg ctccataaca
     3481 tcaaacatcg acccacggcg taacgcgctt gctgcttgga tgcccgaggc atagactgta
     3541 caaaaaaaca gtcataacaa gccatgaaaa ccgccactgc gccgttacca ccgctgcgtt
     3601 cggtcaaggt tctggaccag ttgcgtgagc gcatacgcta cttgcattac agtttacgaa
     3661 ccgaacaggc ttatgtcaac tgggttcgtg ccttcatccg tttccacggt gtgcgtcacc
     3721 cggcaacctt gggcagcagc gaagtcgagg catttctgtc ctggctggcg aacgagcgca
     3781 aggtttcggt ctccacgcat cgtcaggcat tggcggcctt gctgttcttc tacggcaagg
     3841 tgctgtgcac ggatctgccc ttgcttcagg agatcggtag acctcggccg tcgcggcgct
     3901 tgccggtggt gctgaccccg gatgaagtgg ttcgcatcct cggttttctg gaaggcgagc
     3961 atcgtttgtt cgcccaggac tctagctata gttctagtgg ttggctacag ctttgtttgt
     4021 actatcaaca ggttgaactg ctgatcaaca gatcctctac gcggccgcgg taccataact
     4081 tcgtatagca tacattatac gaagttatct ggtttaaacg tacccgtagt ggctatggca
     4141 gggcttgccg ccccgacgtt ggctgcgagc cctgggcctt cacccgaact tgggggttgg
     4201 ggtggggaaa aggaagaaac gcgggcgtat tggtcccaat ggggtctcgg tggggtatcg
     4261 acagagtgcc agccctggga ccgaaccccg cgtttatgaa caaacgaccc aacacccgtg
     4321 cgttttattc tgtcttttta ttgccgtcat agcgcgggtt ccttccggta ttgtctcctt
     4381 ccgtgtttca gttagcctcc cccatctccc ggtaccgcat gctatgcatc agctgctagc
     4441 accatggctc gagatcccgg gtgatcaagt cttcgtcgag tgattgtaaa taaaatgtaa
     4501 tttacagtat agtattttaa ttaatataca aatgatttga taataattct tatttaacta
     4561 taatatattg tgttgggttg aattaaaggt ccgtatacta gtatcgattc gcgacctact
     4621 ccggaatatt aatagatcat ggagataatt aaaatgataa ccatctcgca aataaataag
     4681 tattttactg ttttcgtaac agttttgtaa taaaaaaacc tataaatatt ccggattatt
     4741 cataccgtcc caccatcggg cgcggatccc ggtccgaagc gcgcggaatt caaaggccta
     4801 cgtcgacgag ctcactagtc gcggccgctt tcgaatctag agcctgcagt ctcgacaagc
     4861 ttgtcgagaa gtactagagg atcataatca gccataccac atttgtagag gttttacttg
     4921 ctttaaaaaa cctcccacac ctc
//
