LOCUS       sequence-316895-        2991 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-term CBP 3C cleavable Amp resistant bacterial expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2991)
  AUTHORS   Altmannova V, Blaha A, Astrinidis S, Reichle H, Weir JR
  TITLE     InteBac - An integrated bacterial and baculovirus expression vector 
            suite.
  JOURNAL   Protein Sci. 2020 Sep 21. doi: 10.1002/pro.3957.
  PUBMED    32955754
REFERENCE   2  (bases 1 to 2991)
  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"; journalName: "Protein 
            Sci. 2020 Sep 21. doi: 10.1002/pro.3957."
COMMENT     Sequence Label: 13 pCOLI_A_CBP
FEATURES             Location/Qualifiers
     source          1..2991
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     171..190
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     424..441
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    456..595
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(521..540)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     681..703
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(697..888)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1125..1144
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1125..1143
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1181..1203
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1214..1291
                     /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             1304..1327
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(1520..1538)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1534..1581
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1646..1737
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(2673..2991,1..270)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
       61 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
      121 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
      181 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
      241 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
      301 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
      361 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
      421 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
      481 catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg
      541 catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg
      601 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
      661 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
      721 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
      781 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
      841 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt
      901 gtaaggggga tttctgttca tggggtaatg ataccgatga aacgagagag gatgctcacg
      961 atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg
     1021 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt
     1081 aatacagatg taggtgttgg caccatgcat cgatcccgcg aaattaatac gactcactat
     1141 agggagacca caacggtttc cctccggctc cggaaataat tttgtttaac tttaagaagg
     1201 agatatatat atgaagcgac gatggaaaaa gaatttcata gccgtctcag cagccaaccg
     1261 ctttaagaaa atctcatcct ccggggcact tagcagcgga tctctggaag ttctgttcca
     1321 ggggcccgga tcccggtccg acatatgagc gcgcggaatt caaaggccta cgtcgacgag
     1381 ctcactagtc gcggccgctt tcgaatctag agcctgcagt ctcgacaagc ttgtcgagaa
     1441 gtactagagg atgtacaacg cgtgcgcgct gatccggctg ctaacaaagc ccgaaaggaa
     1501 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
     1561 cgggtcttga ggggtttttt gctgaaagga ggaactatat cgggattagc ccgcctaatg
     1621 agcgggcttt tttttaattc ccctatttgt ttatttttct aaatacattc aaatatgtat
     1681 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     1741 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     1801 tttgctcacc cagaaacgct cgtgaaagta aaagacgcag aggaccaatt gggggcacga
     1861 gtgggataca tagaactgga cttgaatagc ggtaaaatcc ttgagagttt tcgccctgaa
     1921 gagcgttttc caatgatgag cactttcaaa gttctgctat gtggagcagt attatcccgt
     1981 gtagatgcgg ggcaagagca actcggacga cgaatacact attcgcagaa tgacttggtt
     2041 gaatactccc cagtgacaga aaagcacctt acggacggaa tgacggtaag agaattatgt
     2101 agtgccgcca taacgatgag tgataacact gcggcgaact tacttctgac aaccatcggt
     2161 ggaccgaagg aattaaccgc ttttttgcac aatatgggag accatgtaac tcgccttgac
     2221 cgttgggaac cagaactgaa tgaagccata ccaaacgacg agcgagacac cacaatgcct
     2281 gcggcaatgg caacaacatt acgcaaacta ttaactggcg aactacttac tctggcttca
     2341 cggcaacaat taatagactg gcttgaagcg gataaagttg caggaccact actgcgttcg
     2401 gcacttcctg ctggctggtt tattgctgat aaatctgggg caggagagcg tggttcacgg
     2461 ggtatcattg ccgcacttgg accagatggt aagccttccc gtatcgtagt tatctacacg
     2521 acgggtagtc aggcaactat ggacgaacga aatagacaga ttgctgaaat aggggcttca
     2581 ctgattaagc attggtaaac cgatacaatt aaaggctcct tttggagcct tttttttttg
     2641 gacggaccgg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     2701 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     2761 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     2821 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     2881 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     2941 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg a
//
