LOCUS       sequence-232375-        7326 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with N-terminal MBP tag, cleavable with 3C, 
            and C-terminal monomeric GFP and HIS6, cleavable with TEV.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7326)
  AUTHORS   Lemaitre RP, Bogdanova A, Borgonovo B, Woodruff JB, Drechsel DN
  TITLE     FlexiBAC: a versatile, open-source baculovirus vector system for 
            protein expression, secretion, and proteolytic processing.
  JOURNAL   BMC Biotechnol. 2019 Mar 29;19(1):20. doi: 
            10.1186/s12896-019-0512-z.
  PUBMED    30925874
REFERENCE   2  (bases 1 to 7326)
  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: "BMC 
            Biotechnol."; date: "2019-03-29"; volume: "19(1):20. doi"; pages: " 
            10.1186/s12896-019-0512-z"
COMMENT     Sequence Label: pOCC120
FEATURES             Location/Qualifiers
     source          1..7326
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      42..51
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             54..1154
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELVKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     1128..1151
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1209..1232
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1341..1646
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1525..1544
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             join(1714..1716,1717..1719,1720..2430)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1759..1780)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2020..2039)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2367..2388
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2431..2454
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2464..2481
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2623..2757
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2660..2679)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2714..2733
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           2830..3202
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(3554..3571)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3664..4252)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3744..3763)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4423..5214,5215..5283))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5046..5065
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5284..5388)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5689..5711
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5703..5720
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5704..5720
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5731..5750
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5731..5749
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6360..6484
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(6522..7127)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     promoter        7235..7326
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     7251..7268
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
ORIGIN
        1 attccggatt attcataccg tcccaccatc gggcgcggat cgccaccatg ggcatgaaaa
       61 tcgaagaagg taaactggta atctggatta acggcgataa aggctataac ggtctcgctg
      121 aagtcggtaa gaaattcgag aaagataccg gaattaaagt caccgttgag catccggata
      181 aactggaaga gaaattccca caggttgcgg caactggcga tggccctgac attatcttct
      241 gggcacacga ccgctttggt ggctacgctc aatctggcct gttggctgaa atcaccccgg
      301 acaaagcgtt ccaggacaag ctgtatccgt ttacctggga tgccgtacgt tacaacggca
      361 agctgattgc ttacccgatc gctgttgaag cgttatcgct gatttataac aaagatctgc
      421 tgccgaaccc gccaaaaacc tgggaagaga tcccggcgct ggataaagaa ctgaaagcga
      481 aaggtaagag cgcgctgatg ttcaacctgc aagaaccgta cttcacctgg ccgctgattg
      541 ctgctgacgg gggttatgcg ttcaagtatg aaaacggcaa gtacgacatt aaagacgtgg
      601 gcgtggataa cgctggcgcg aaagcgggtc tgaccttcct ggttgacctg attaaaaaca
      661 aacacatgaa tgcagacacc gattactcca tcgcagaagc tgcctttaat aaaggcgaaa
      721 cagcgatgac catcaacggc ccgtgggcat ggtccaacat cgacaccagc aaagtgaatt
      781 atggtgtaac ggtactgccg accttcaagg gtcaaccatc caaaccgttc gttggcgtgc
      841 tgagcgcagg tattaacgcc gccagtccga acaaagagct ggcaaaagag ttcctcgaaa
      901 actatctgct gactgatgaa ggtctggaag cggttaataa agacaaaccg ctgggtgccg
      961 tagcgctgaa gtcttacgag gaagagttgg tgaaagatcc gcgtattgcc gccactatgg
     1021 aaaacgccca gaaaggtgaa atcatgccga acatcccgca gatgtccgct ttctggtatg
     1081 ccgtgcgtac tgcggtgatc aacgccgcca gcggtcgtca gactgtcgat gaagccctga
     1141 aagacgcgca gactaattcg agctcgaaca acaacaacaa taacaataac aacaacagca
     1201 gcggcagact ggaagttctg ttccaggggc cggcggccgc aagcgctatg gccacaacct
     1261 gttacattgc acaagataaa aatatatcat cacgaacagt aaaactgtct gcttacataa
     1321 acagtaatac aaggggtgtt atgcagttta aggtttacac ctataaaaga gagagccgtt
     1381 atcgtctgtt tgtggatgta cagagtgata ttattgacac gcccgggcga cggatggtga
     1441 tccccctggc cagtgcccgt ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg
     1501 tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga tatggccagt gtgccggtct
     1561 ccgttatcgg ggaagaagtg gctgatctca gccaccgcga aaatgacatc aaaaacgcca
     1621 ttaacctgat gttctgggga atataaatgt caggctccct tatacacagc cagtggcgcg
     1681 ccgggatccg ctggctccgc tgctggttct ggcatggtga gcaagggcga ggagctgttc
     1741 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     1801 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1861 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1921 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1981 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     2041 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     2101 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     2161 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     2221 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     2281 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc caaactgagc
     2341 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     2401 atcactctcg gcatggacga gctgtacaag ctggaagttc tgttccaggg gccgggcagc
     2461 agccatcacc atcaccatca cagtggataa tagaagcttg tcgagaagta ctagaggatc
     2521 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     2581 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     2641 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     2701 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatctga
     2761 tcactgcttg agcctaggag atccgaacca gataagtgaa agcctgcatc cgcagatctt
     2821 aattaataaa acacgataca ttgttattag tacatttatt aagcgctaga ttctgtgcgt
     2881 tgttgattta cagacaattg ttgtacgtat tttaataatt cattaaattt ataatcttta
     2941 gggtggtatg ttagagcgaa aatcaaatga ttttcagcgt ctttatatct gaatttaaat
     3001 attaaatcct caatagattt gtaaaatagg tttcgattag tttcaaacaa gggttgtttt
     3061 tccgaaccga tggctggact atctaatgga ttttcgctca acgccacaaa acttgccaaa
     3121 tcttgtagca gcaatctagc tttgtcgata ttcgtttgtg ttttgttttg taataaaggt
     3181 tcgacgtcgt tcaaaatatt atgcgctttt gtatttcttt catcactgtc gttagtgtac
     3241 aattgactcg acgtaaacac gttaaataac gcttggacat atttaacatc gggcgtgtta
     3301 gctttattag gccgattatc gtcgtcgtcc caaccctcgt cgttagaagt tgcttccgaa
     3361 gacgattttg ccatagccac acgacgccta ttaattgtgt cggctaacac gtccgcgatc
     3421 aaatttgtag ttgagctttt tgggcctcat gggccttccg ctcactgccc gctttccagt
     3481 cgggaaacct gtcgtgccag ctgcattaac atggtcatag ctgtttcctt gcgtattggg
     3541 cgctctccgc ttcctcgctc actgactcgc tgcgctcggt cgttcgggta aagcctgggg
     3601 tgcctaatga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3661 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3721 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3781 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3841 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3901 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3961 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4021 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4081 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4141 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4201 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4261 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4321 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4381 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4441 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4501 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4561 gcgagaacca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4621 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4681 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4741 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4801 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4861 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4921 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4981 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     5041 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5101 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5161 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5221 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5281 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5341 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     5401 aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     5461 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     5521 gaatagaccg agatagggtt gagtggccgc tacagggcgc tcccattcgc cattcaggct
     5581 gcgcaactgt tgggaagggc gtttcggtgc gggcctcttc gctattacgc cagctggcga
     5641 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     5701 gttgtaaaac gacggccagt gagcgcgacg taatacgact cactataggg cgaattggcg
     5761 gaaggccgtc aaggccgtga cgttaaaact attaagccat ccaatcgacc gttagtcgaa
     5821 tcaggaccgc tggtgcgaga agccgcgaag tatggcgaat gcatcgtata acgtgtggag
     5881 tccgctcatt agagcgtcat gtttagacaa gaaagctaca tatttaattg atcccgatga
     5941 ttttattgat aaattgaccc taactccata cacggtattc tacaatggcg gggttttggt
     6001 caaaatttcc ggactgcgat tgtacatgct gttaacggct ccgcccacta ttaatgaaat
     6061 taaaaattcc aattttaaaa aacgcagcaa gagaaacatt tgtatgaaag aatgcgtaga
     6121 aggaaagaaa aatgtcgtgg acatgctgaa caacaagatt aatatgcctc cgtgtataaa
     6181 aaaaatattg aacgatttga aagaaaacaa tgtaccgcgc ggcggtatgt acaggaagag
     6241 gtttatacta aactgttaca ttgcaaacgt ggtttcgtgt gccaagtgtg aaaaccgatg
     6301 tttaatcaag gctctgacgc atttctacaa ccacgactcc aagtgtgtgg gtgaagtcat
     6361 gcatctttta atcaaatccc aagatgtgta taaaccacca aactgccaaa aaatgaaaac
     6421 tgtggacaag ctctgtccgt ttgctggcaa ctgcaagggt ctcaatccta tttgtaatta
     6481 ttgaataata aaacaattat aaatgctaaa tttgtttttt attaacgata caaaccaaac
     6541 gcaacaagaa catttgtagt attatctata attgaaaacg cgtagttata atcgctgagg
     6601 taatatttaa aatcattttc aaatgattca cagttaattt gcgacaatat aattttattt
     6661 tcacataaac tagacgcctt gtcgtcttct tcttcgtatt ccttctcttt ttcatttttc
     6721 tcctcataaa aattaacata gttattatcg tatccatata tgtatctatc gtatagagta
     6781 aattttttgt tgtcataaat atatatgtct tttttaatgg ggtgtatagt accgctgcgc
     6841 atagtttttc tgtaatttac aacagtgcta ttttctggta gttcttcgga gtgtgttgct
     6901 ttaattatta aatttatata atcaatgaat ttgggatcgt cggttttgta caatatgttg
     6961 ccggcatagt acgcagcttc ttctagttca attacaccat tttttagcag caccggatta
     7021 acataacttt ccaaaatgtt gtacgaaccg ttaaacaaaa acagttcacc tcccttttct
     7081 atactattgt ctgcgagcag ttgtttgttg ttaaaaataa cagccattgt aatgagacgc
     7141 acaaactaat atcacaagct gccaccattt tgttcgccca ggactctagc tatagttcta
     7201 gtggttggct acgtatactc cggaatatta atagatcatg gagataatta aaatgataac
     7261 catctcgcaa ataaataagt attttactgt tttcgtaaca gttttgtaat aaaaaaacct
     7321 ataaat
//
