LOCUS       sequence-232906-        3563 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Plasmid for expression of C-terminally CBP-3xFLAG-ProteinA-tagged 
            proteins under the control of a polh promoter in baculovirus/insect 
            cell systems.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3563)
  AUTHORS   Monika Golas
  TITLE     Plasmids for expression of proteins in insect cells
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3563)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pGS-BacA-21232
FEATURES             Location/Qualifiers
     source          1..3563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      190..778
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     679..698
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     mobile_element  913..1136
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(1203..1736)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1925..1930,1931..1947,1948..1953))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(2024..2057)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2134..2225
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     2150..2167
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             2351..2428
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             2444..2509
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2537..2557
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2597..2770
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             2771..2944
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNGAQAPK"
     polyA_signal    3143..3277
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3180..3199)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3234..3253
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3398..3563
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 accggttgac ttgggtcaac tgtcagacca agtttactca tatatacttt agattgattt
       61 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
      121 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
      181 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
      241 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
      301 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
      361 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
      421 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
      481 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
      541 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
      601 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
      661 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
      721 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
      781 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
      841 ctttcctgcg ttatcccctg attgacttgg gtcgctcttc ctgtggatgc gcagatgccc
      901 tgcgtaagcg ggtgtgggcg gacaataaag tcttaaactg aacaaaatag atctaaacta
      961 tgacaataaa gtcttaaact agacagaata gttgtaaact gaaatcagtc cagttatgct
     1021 gtgaaaaagc atactggact tttgttatgg ctaaagcaaa ctcttcattt tctgaagtgc
     1081 aaattgcccg tcgtattaaa gaggggcgtg gccaagggca tgtaaagact atattcgcgg
     1141 cgttgtgaca atttaccgaa caactccgcg gccgggaagc cgatctcggc ttgaacgaat
     1201 tgttaggtgg cggtacttgg gtcgatatca aagtgcatca cttcttcccg tatgcccaac
     1261 tttgtataga gagccactgc gggatcgtca ccgtaatctg cttgcacgta gatcacataa
     1321 gcaccaagcg cgttggcctc atgcttgagg agattgatga gcgcggtggc aatgccctgc
     1381 ctccggtgct cgccggagac tgcgagatca tagatataga tctcactacg cggctgctca
     1441 aacttgggca gaacgtaagc cgcgagagcg ccaacaaccg cttcttggtc gaaggcagca
     1501 agcgcgatga atgtcttact acggagcaag ttcccgaggt aatcggagtc cggctgatgt
     1561 tgggagtagg tggctacgtc tccgaactca cgaccgaaaa gatcaagagc agcccgcatg
     1621 gatttgactt ggtcagggcc gagcctacat gtgcgaatga tgcccatact tgagccacct
     1681 aactttgttt tagggcgact gccctgctgc gtaacatcgt tgctgctgcg taacatcgtt
     1741 gctgctccat aacatcaaac atcgacccac ggcgtaacgc gcttgctgct tggatgcccg
     1801 aggcatagac tgtacaaaaa aacagtcata acaagccatg aaaaccgcca ctgcgccgtt
     1861 accaccgctg cgttcggtca aggttctgga ccagttgcgt gagcgcatac gctacttgca
     1921 ttacagttta cgaaccgaac aggcttatgt caactgggtt cgtgccttca tccgtttcca
     1981 cggtgtgcgt cacccggcaa ccttgggcag cagcgaagtc gccataactt cgtatagcat
     2041 acattatacg aagttatctg taactataac ggtcctaagg tagcgagttt aaacactagt
     2101 atcgattcgc gacctactcc ggaatattaa tagatcatgg agataattaa aatgataacc
     2161 atctcgcaaa taaataagta ttttactgtt ttcgtaacag ttttgtaata aaaaaaccta
     2221 taaatattcc ggattattca taccgtccca ccatcgggcg cggatccgtc acggtcgacg
     2281 taacgggtac ccgtagttaa ttacggcgcg ccgtaacgac gtccctgtac tcgagggttc
     2341 tggttctggt aaaagaagat ggaagaagaa ttttattgct gtttctgctg ctaatagatt
     2401 taagaaaatt tcttcttctg gtgctttgtc tgcttctggt tctgactaca aagaccatga
     2461 cggtgattat aaagatcatg acatcgatta caaggatgac gatgacaagg attatgatat
     2521 tccaactact gctagcgaga atttgtattt tcagggtgag ctcaaaaccg cggctcttgc
     2581 gcaacacgat gaagccgtgg acaacaaatt caacaaagaa caacaaaacg cgttctatga
     2641 gatcttacat ttacctaact taaacgaaga acaacgaaac gccttcatcc aaagtttaaa
     2701 agatgaccca agccaaagcg ctaacctttt agcagaagct aaaaagctaa atgatgctca
     2761 ggcgccgaaa gtagacaaca aattcaacaa agaacaacaa aacgcgttct atgagatctt
     2821 acatttacct aacttaaacg aagaacaacg aaacgccttc atccaaagtt taaaagatga
     2881 cccaagccaa agcgctaacc ttttagcaga agctaaaaag ctaaatggtg ctcaggcgcc
     2941 gaaagtagac gcgaattccg cggggaagtc aacctgagcg gccgctttcg aatctagagc
     3001 ctgcagtctc gacaagcttg tcgagaagta ctagaggatc ataatcagcc ataccacatt
     3061 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     3121 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     3181 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     3241 gtccaaactc atcaatgtat cttatcatgt ctggatctga tcactgcttg agcctagaag
     3301 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
     3361 aactatcata acccctaggg tatacccatc taattggaac cagataagtg aaatctagtt
     3421 ccaaactatt ttgtcatttt taattttcgt attagcttac gacgctacac ccagttccca
     3481 tctattttgt cactcttccc taaataatcc ttaaaaactc catttccacc cctcccagtt
     3541 cccaactatt ttgtccgccc aca
//
