LOCUS       sequence-232311-        7164 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for rec. protein with N-terminal MBP tag, cleavable with 3C and 
            C-terminal SNAP, cleavable with TEV and HIS6, cleavable with 3C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7164)
  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 7164)
  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: pOCC210
FEATURES             Location/Qualifiers
     source          1..7164
                     /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             1702..1722
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1723..2268
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     CDS             2269..2292
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2302..2319
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2461..2595
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2498..2517)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2552..2571
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           2668..3040
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(3392..3409)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3502..4090)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3582..3601)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4261..5052,5053..5121))
                     /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     4884..4903
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5122..5226)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5527..5549
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5541..5558
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5542..5558
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5569..5588
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5569..5587
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6198..6322
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(6360..6965)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     promoter        7073..7164
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     7089..7106
                     /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 ccgggatcca gcagcggcag agaaaacttg tatttccagg gcatggacaa agactgcgaa
     1741 atgaagcgca ccaccctgga tagccctctg ggcaagctgg aactgtctgg gtgcgaacag
     1801 ggcctgcacc gtatcatctt cctgggcaaa ggaacatctg ccgccgacgc cgtggaagtg
     1861 cctgccccag ccgccgtgct gggcggacca gagccactga tgcaggccac cgcctggctc
     1921 aacgcctact ttcaccagcc tgaggccatc gaggagttcc ctgtgccagc cctgcaccac
     1981 ccagtgttcc agcaggagag ctttacccgc caggtgctgt ggaaactgct gaaagtggtg
     2041 aagttcggag aggtcatcag ctacagccac ctggccgccc tggccggcaa tcccgccgcc
     2101 accgccgccg tgaaaaccgc cctgagcgga aatcccgtgc ccattctgat cccctgccac
     2161 cgggtggtgc agggcgacct ggacgtgggg ggctacgagg gcgggctcgc cgtgaaagag
     2221 tggctgctgg cccacgaggg ccacagactg ggcaagcctg ggctgggtct ggaagttctg
     2281 ttccaggggc cgggcagcag ccatcaccat caccatcaca gtggataata gaagcttgtc
     2341 gagaagtact agaggatcat aatcagccat accacatttg tagaggtttt acttgcttta
     2401 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     2461 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     2521 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     2581 tatcatgtct ggatctgatc actgcttgag cctaggagat ccgaaccaga taagtgaaag
     2641 cctgcatccg cagatcttaa ttaataaaac acgatacatt gttattagta catttattaa
     2701 gcgctagatt ctgtgcgttg ttgatttaca gacaattgtt gtacgtattt taataattca
     2761 ttaaatttat aatctttagg gtggtatgtt agagcgaaaa tcaaatgatt ttcagcgtct
     2821 ttatatctga atttaaatat taaatcctca atagatttgt aaaataggtt tcgattagtt
     2881 tcaaacaagg gttgtttttc cgaaccgatg gctggactat ctaatggatt ttcgctcaac
     2941 gccacaaaac ttgccaaatc ttgtagcagc aatctagctt tgtcgatatt cgtttgtgtt
     3001 ttgttttgta ataaaggttc gacgtcgttc aaaatattat gcgcttttgt atttctttca
     3061 tcactgtcgt tagtgtacaa ttgactcgac gtaaacacgt taaataacgc ttggacatat
     3121 ttaacatcgg gcgtgttagc tttattaggc cgattatcgt cgtcgtccca accctcgtcg
     3181 ttagaagttg cttccgaaga cgattttgcc atagccacac gacgcctatt aattgtgtcg
     3241 gctaacacgt ccgcgatcaa atttgtagtt gagctttttg ggcctcatgg gccttccgct
     3301 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaacat ggtcatagct
     3361 gtttccttgc gtattgggcg ctctccgctt cctcgctcac tgactcgctg cgctcggtcg
     3421 ttcgggtaaa gcctggggtg cctaatgagc aaaaggccag caaaaggcca ggaaccgtaa
     3481 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3541 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3601 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3661 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3721 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3781 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3841 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3901 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3961 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4021 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4081 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4141 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4201 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4261 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4321 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4381 cagtgctgca atgataccgc gagaaccacg ctcaccggct ccagatttat cagcaataaa
     4441 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4501 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4561 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4621 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4681 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4741 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4801 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4861 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4921 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4981 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5041 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5101 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5161 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5221 tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa
     5281 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     5341 atcccttata aatcaaaaga atagaccgag atagggttga gtggccgcta cagggcgctc
     5401 ccattcgcca ttcaggctgc gcaactgttg ggaagggcgt ttcggtgcgg gcctcttcgc
     5461 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     5521 ggttttccca gtcacgacgt tgtaaaacga cggccagtga gcgcgacgta atacgactca
     5581 ctatagggcg aattggcgga aggccgtcaa ggccgtgacg ttaaaactat taagccatcc
     5641 aatcgaccgt tagtcgaatc aggaccgctg gtgcgagaag ccgcgaagta tggcgaatgc
     5701 atcgtataac gtgtggagtc cgctcattag agcgtcatgt ttagacaaga aagctacata
     5761 tttaattgat cccgatgatt ttattgataa attgacccta actccataca cggtattcta
     5821 caatggcggg gttttggtca aaatttccgg actgcgattg tacatgctgt taacggctcc
     5881 gcccactatt aatgaaatta aaaattccaa ttttaaaaaa cgcagcaaga gaaacatttg
     5941 tatgaaagaa tgcgtagaag gaaagaaaaa tgtcgtggac atgctgaaca acaagattaa
     6001 tatgcctccg tgtataaaaa aaatattgaa cgatttgaaa gaaaacaatg taccgcgcgg
     6061 cggtatgtac aggaagaggt ttatactaaa ctgttacatt gcaaacgtgg tttcgtgtgc
     6121 caagtgtgaa aaccgatgtt taatcaaggc tctgacgcat ttctacaacc acgactccaa
     6181 gtgtgtgggt gaagtcatgc atcttttaat caaatcccaa gatgtgtata aaccaccaaa
     6241 ctgccaaaaa atgaaaactg tggacaagct ctgtccgttt gctggcaact gcaagggtct
     6301 caatcctatt tgtaattatt gaataataaa acaattataa atgctaaatt tgttttttat
     6361 taacgataca aaccaaacgc aacaagaaca tttgtagtat tatctataat tgaaaacgcg
     6421 tagttataat cgctgaggta atatttaaaa tcattttcaa atgattcaca gttaatttgc
     6481 gacaatataa ttttattttc acataaacta gacgccttgt cgtcttcttc ttcgtattcc
     6541 ttctcttttt catttttctc ctcataaaaa ttaacatagt tattatcgta tccatatatg
     6601 tatctatcgt atagagtaaa ttttttgttg tcataaatat atatgtcttt tttaatgggg
     6661 tgtatagtac cgctgcgcat agtttttctg taatttacaa cagtgctatt ttctggtagt
     6721 tcttcggagt gtgttgcttt aattattaaa tttatataat caatgaattt gggatcgtcg
     6781 gttttgtaca atatgttgcc ggcatagtac gcagcttctt ctagttcaat tacaccattt
     6841 tttagcagca ccggattaac ataactttcc aaaatgttgt acgaaccgtt aaacaaaaac
     6901 agttcacctc ccttttctat actattgtct gcgagcagtt gtttgttgtt aaaaataaca
     6961 gccattgtaa tgagacgcac aaactaatat cacaagctgc caccattttg ttcgcccagg
     7021 actctagcta tagttctagt ggttggctac gtatactccg gaatattaat agatcatgga
     7081 gataattaaa atgataacca tctcgcaaat aaataagtat tttactgttt tcgtaacagt
     7141 tttgtaataa aaaaacctat aaat
//
