LOCUS       sequence-224663-        6580 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 HIS6 tag, cleavable with 3C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6580)
  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 6580)
  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: pOCC28
FEATURES             Location/Qualifiers
     source          1..6580
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        79..170
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     95..112
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     regulatory      212..221
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             224..1324
                     /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     1298..1321
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1379..1402
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1511..1816
                     /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     1695..1714
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             1854..1877
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1887..1904
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2046..2180
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2083..2102)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2137..2156
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           2253..2625
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(2977..2994)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3087..3675)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3167..3186)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3846..4637,4638..4706))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4469..4488
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4708..4812)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5113..5135
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5127..5144
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5128..5144
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5155..5174
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5155..5173
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5784..5908
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5946..6551)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
ORIGIN
        1 agctgccacc attttgttcg cccaggactc tagctatagt tctagtggtt ggctacgtat
       61 actccggaat attaatagat catggagata attaaaatga taaccatctc gcaaataaat
      121 aagtatttta ctgttttcgt aacagttttg taataaaaaa acctataaat attccggatt
      181 attcataccg tcccaccatc gggcgcggat cgccaccatg ggcatgaaaa tcgaagaagg
      241 taaactggta atctggatta acggcgataa aggctataac ggtctcgctg aagtcggtaa
      301 gaaattcgag aaagataccg gaattaaagt caccgttgag catccggata aactggaaga
      361 gaaattccca caggttgcgg caactggcga tggccctgac attatcttct gggcacacga
      421 ccgctttggt ggctacgctc aatctggcct gttggctgaa atcaccccgg acaaagcgtt
      481 ccaggacaag ctgtatccgt ttacctggga tgccgtacgt tacaacggca agctgattgc
      541 ttacccgatc gctgttgaag cgttatcgct gatttataac aaagatctgc tgccgaaccc
      601 gccaaaaacc tgggaagaga tcccggcgct ggataaagaa ctgaaagcga aaggtaagag
      661 cgcgctgatg ttcaacctgc aagaaccgta cttcacctgg ccgctgattg ctgctgacgg
      721 gggttatgcg ttcaagtatg aaaacggcaa gtacgacatt aaagacgtgg gcgtggataa
      781 cgctggcgcg aaagcgggtc tgaccttcct ggttgacctg attaaaaaca aacacatgaa
      841 tgcagacacc gattactcca tcgcagaagc tgcctttaat aaaggcgaaa cagcgatgac
      901 catcaacggc ccgtgggcat ggtccaacat cgacaccagc aaagtgaatt atggtgtaac
      961 ggtactgccg accttcaagg gtcaaccatc caaaccgttc gttggcgtgc tgagcgcagg
     1021 tattaacgcc gccagtccga acaaagagct ggcaaaagag ttcctcgaaa actatctgct
     1081 gactgatgaa ggtctggaag cggttaataa agacaaaccg ctgggtgccg tagcgctgaa
     1141 gtcttacgag gaagagttgg tgaaagatcc gcgtattgcc gccactatgg aaaacgccca
     1201 gaaaggtgaa atcatgccga acatcccgca gatgtccgct ttctggtatg ccgtgcgtac
     1261 tgcggtgatc aacgccgcca gcggtcgtca gactgtcgat gaagccctga aagacgcgca
     1321 gactaattcg agctcgaaca acaacaacaa taacaataac aacaacagca gcggcagact
     1381 ggaagttctg ttccaggggc cggcggccgc aagcgctatg gccacaacct gttacattgc
     1441 acaagataaa aatatatcat cacgaacagt aaaactgtct gcttacataa acagtaatac
     1501 aaggggtgtt atgcagttta aggtttacac ctataaaaga gagagccgtt atcgtctgtt
     1561 tgtggatgta cagagtgata ttattgacac gcccgggcga cggatggtga tccccctggc
     1621 cagtgcccgt ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg
     1681 ggatgaaagc tggcgcatga tgaccaccga tatggccagt gtgccggtct ccgttatcgg
     1741 ggaagaagtg gctgatctca gccaccgcga aaatgacatc aaaaacgcca ttaacctgat
     1801 gttctgggga atataaatgt caggctccct tatacacagc cagtggcgcg ccgctggaag
     1861 ttctgttcca ggggccgggc agcagccatc accatcacca tcacagtgga taatagaagc
     1921 ttgtcgagaa gtactagagg atcataatca gccataccac atttgtagag gttttacttg
     1981 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     2041 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     2101 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     2161 tatcttatca tgtctggatc tgatcactgc ttgagcctag gagatccgaa ccagataagt
     2221 gaaagcctgc atccgcagat cttaattaat aaaacacgat acattgttat tagtacattt
     2281 attaagcgct agattctgtg cgttgttgat ttacagacaa ttgttgtacg tattttaata
     2341 attcattaaa tttataatct ttagggtggt atgttagagc gaaaatcaaa tgattttcag
     2401 cgtctttata tctgaattta aatattaaat cctcaataga tttgtaaaat aggtttcgat
     2461 tagtttcaaa caagggttgt ttttccgaac cgatggctgg actatctaat ggattttcgc
     2521 tcaacgccac aaaacttgcc aaatcttgta gcagcaatct agctttgtcg atattcgttt
     2581 gtgttttgtt ttgtaataaa ggttcgacgt cgttcaaaat attatgcgct tttgtatttc
     2641 tttcatcact gtcgttagtg tacaattgac tcgacgtaaa cacgttaaat aacgcttgga
     2701 catatttaac atcgggcgtg ttagctttat taggccgatt atcgtcgtcg tcccaaccct
     2761 cgtcgttaga agttgcttcc gaagacgatt ttgccatagc cacacgacgc ctattaattg
     2821 tgtcggctaa cacgtccgcg atcaaatttg tagttgagct ttttgggcct catgggcctt
     2881 ccgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aacatggtca
     2941 tagctgtttc cttgcgtatt gggcgctctc cgcttcctcg ctcactgact cgctgcgctc
     3001 ggtcgttcgg gtaaagcctg gggtgcctaa tgagcaaaag gccagcaaaa ggccaggaac
     3061 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3121 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3181 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3241 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3301 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3361 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3421 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3481 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     3541 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3601 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3661 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3721 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3781 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     3841 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     3901 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     3961 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4021 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4081 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4141 cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4201 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4261 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4321 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4381 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4441 agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa
     4501 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4561 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4621 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4681 gcgacacgga aatgttgaat actcattact cttccttttt caatattatt gaagcattta
     4741 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     4801 aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg
     4861 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     4921 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgg ccgctacagg
     4981 gcgctcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgtttcg gtgcgggcct
     5041 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     5101 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc gacgtaatac
     5161 gactcactat agggcgaatt ggcggaaggc cgtcaaggcc gtgacgttaa aactattaag
     5221 ccatccaatc gaccgttagt cgaatcagga ccgctggtgc gagaagccgc gaagtatggc
     5281 gaatgcatcg tataacgtgt ggagtccgct cattagagcg tcatgtttag acaagaaagc
     5341 tacatattta attgatcccg atgattttat tgataaattg accctaactc catacacggt
     5401 attctacaat ggcggggttt tggtcaaaat ttccggactg cgattgtaca tgctgttaac
     5461 ggctccgccc actattaatg aaattaaaaa ttccaatttt aaaaaacgca gcaagagaaa
     5521 catttgtatg aaagaatgcg tagaaggaaa gaaaaatgtc gtggacatgc tgaacaacaa
     5581 gattaatatg cctccgtgta taaaaaaaat attgaacgat ttgaaagaaa acaatgtacc
     5641 gcgcggcggt atgtacagga agaggtttat actaaactgt tacattgcaa acgtggtttc
     5701 gtgtgccaag tgtgaaaacc gatgtttaat caaggctctg acgcatttct acaaccacga
     5761 ctccaagtgt gtgggtgaag tcatgcatct tttaatcaaa tcccaagatg tgtataaacc
     5821 accaaactgc caaaaaatga aaactgtgga caagctctgt ccgtttgctg gcaactgcaa
     5881 gggtctcaat cctatttgta attattgaat aataaaacaa ttataaatgc taaatttgtt
     5941 ttttattaac gatacaaacc aaacgcaaca agaacatttg tagtattatc tataattgaa
     6001 aacgcgtagt tataatcgct gaggtaatat ttaaaatcat tttcaaatga ttcacagtta
     6061 atttgcgaca atataatttt attttcacat aaactagacg ccttgtcgtc ttcttcttcg
     6121 tattccttct ctttttcatt tttctcctca taaaaattaa catagttatt atcgtatcca
     6181 tatatgtatc tatcgtatag agtaaatttt ttgttgtcat aaatatatat gtctttttta
     6241 atggggtgta tagtaccgct gcgcatagtt tttctgtaat ttacaacagt gctattttct
     6301 ggtagttctt cggagtgtgt tgctttaatt attaaattta tataatcaat gaatttggga
     6361 tcgtcggttt tgtacaatat gttgccggca tagtacgcag cttcttctag ttcaattaca
     6421 ccatttttta gcagcaccgg attaacataa ctttccaaaa tgttgtacga accgttaaac
     6481 aaaaacagtt cacctccctt ttctatacta ttgtctgcga gcagttgttt gttgttaaaa
     6541 ataacagcca ttgtaatgag acgcacaaac taatatcaca
//
