LOCUS       sequence-232245-        7134 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with N-terminal HIS6-MBP tag, cleavable with
            3C, and N-terminal SNAP tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7134)
  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 7134)
  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: pOCC171
FEATURES             Location/Qualifiers
     source          1..7134
                     /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             60..77
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             90..1190
                     /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     1164..1187
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1245..1268
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1269..1814
                     /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             1953..2258
                     /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     2137..2156
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    2431..2565
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2468..2487)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2522..2541
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           2638..3010
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(3362..3379)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3472..4060)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3552..3571)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4231..5022,5023..5091))
                     /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     4854..4873
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5092..5196)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5497..5519
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5511..5528
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5512..5528
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5539..5558
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5539..5557
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6168..6292
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(6330..6935)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     promoter        7043..7134
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     7059..7076
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
ORIGIN
        1 attccggatt attcataccg tcccaccatc gggcgcggat cgccaccatg ggcagcagcc
       61 atcaccatca ccatcacagc agcggcagaa tgaaaatcga agaaggtaaa ctggtaatct
      121 ggattaacgg cgataaaggc tataacggtc tcgctgaagt cggtaagaaa ttcgagaaag
      181 ataccggaat taaagtcacc gttgagcatc cggataaact ggaagagaaa ttcccacagg
      241 ttgcggcaac tggcgatggc cctgacatta tcttctgggc acacgaccgc tttggtggct
      301 acgctcaatc tggcctgttg gctgaaatca ccccggacaa agcgttccag gacaagctgt
      361 atccgtttac ctgggatgcc gtacgttaca acggcaagct gattgcttac ccgatcgctg
      421 ttgaagcgtt atcgctgatt tataacaaag atctgctgcc gaacccgcca aaaacctggg
      481 aagagatccc ggcgctggat aaagaactga aagcgaaagg taagagcgcg ctgatgttca
      541 acctgcaaga accgtacttc acctggccgc tgattgctgc tgacgggggt tatgcgttca
      601 agtatgaaaa cggcaagtac gacattaaag acgtgggcgt ggataacgct ggcgcgaaag
      661 cgggtctgac cttcctggtt gacctgatta aaaacaaaca catgaatgca gacaccgatt
      721 actccatcgc agaagctgcc tttaataaag gcgaaacagc gatgaccatc aacggcccgt
      781 gggcatggtc caacatcgac accagcaaag tgaattatgg tgtaacggta ctgccgacct
      841 tcaagggtca accatccaaa ccgttcgttg gcgtgctgag cgcaggtatt aacgccgcca
      901 gtccgaacaa agagctggca aaagagttcc tcgaaaacta tctgctgact gatgaaggtc
      961 tggaagcggt taataaagac aaaccgctgg gtgccgtagc gctgaagtct tacgaggaag
     1021 agttggtgaa agatccgcgt attgccgcca ctatggaaaa cgcccagaaa ggtgaaatca
     1081 tgccgaacat cccgcagatg tccgctttct ggtatgccgt gcgtactgcg gtgatcaacg
     1141 ccgccagcgg tcgtcagact gtcgatgaag ccctgaaaga cgcgcagact aattcgagct
     1201 cgaacaacaa caacaataac aataacaaca acagcagcgg cagactggaa gttctgttcc
     1261 aggggccgat ggacaaagac tgcgaaatga agcgcaccac cctggatagc cctctgggca
     1321 agctggaact gtctgggtgc gaacagggcc tgcaccgtat catcttcctg ggcaaaggaa
     1381 catctgccgc cgacgccgtg gaagtgcctg ccccagccgc cgtgctgggc ggaccagagc
     1441 cactgatgca ggccaccgcc tggctcaacg cctactttca ccagcctgag gccatcgagg
     1501 agttccctgt gccagccctg caccacccag tgttccagca ggagagcttt acccgccagg
     1561 tgctgtggaa actgctgaaa gtggtgaagt tcggagaggt catcagctac agccacctgg
     1621 ccgccctggc cggcaatccc gccgccaccg ccgccgtgaa aaccgccctg agcggaaatc
     1681 ccgtgcccat tctgatcccc tgccaccggg tggtgcaggg cgacctggac gtggggggct
     1741 acgagggcgg gctcgccgtg aaagagtggc tgctggccca cgagggccac agactgggca
     1801 agcctgggct gggtggatcc gctggctccg ctgctggttc tggcgcggcc gcaagcgcta
     1861 tggccacaac ctgttacatt gcacaagata aaaatatatc atcacgaaca gtaaaactgt
     1921 ctgcttacat aaacagtaat acaaggggtg ttatgcagtt taaggtttac acctataaaa
     1981 gagagagccg ttatcgtctg tttgtggatg tacagagtga tattattgac acgcccgggc
     2041 gacggatggt gatccccctg gccagtgccc gtctgctgtc agataaagtc tcccgtgaac
     2101 tttacccggt ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc gatatggcca
     2161 gtgtgccggt ctccgttatc ggggaagaag tggctgatct cagccaccgc gaaaatgaca
     2221 tcaaaaacgc cattaacctg atgttctggg gaatataaat gtcaggctcc cttatacaca
     2281 gccagtggcg cgccgtaata gaagcttgtc gagaagtact agaggatcat aatcagccat
     2341 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     2401 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     2461 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     2521 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatctgatc actgcttgag
     2581 cctaggagat ccgaaccaga taagtgaaag cctgcatccg cagatcttaa ttaataaaac
     2641 acgatacatt gttattagta catttattaa gcgctagatt ctgtgcgttg ttgatttaca
     2701 gacaattgtt gtacgtattt taataattca ttaaatttat aatctttagg gtggtatgtt
     2761 agagcgaaaa tcaaatgatt ttcagcgtct ttatatctga atttaaatat taaatcctca
     2821 atagatttgt aaaataggtt tcgattagtt tcaaacaagg gttgtttttc cgaaccgatg
     2881 gctggactat ctaatggatt ttcgctcaac gccacaaaac ttgccaaatc ttgtagcagc
     2941 aatctagctt tgtcgatatt cgtttgtgtt ttgttttgta ataaaggttc gacgtcgttc
     3001 aaaatattat gcgcttttgt atttctttca tcactgtcgt tagtgtacaa ttgactcgac
     3061 gtaaacacgt taaataacgc ttggacatat ttaacatcgg gcgtgttagc tttattaggc
     3121 cgattatcgt cgtcgtccca accctcgtcg ttagaagttg cttccgaaga cgattttgcc
     3181 atagccacac gacgcctatt aattgtgtcg gctaacacgt ccgcgatcaa atttgtagtt
     3241 gagctttttg ggcctcatgg gccttccgct cactgcccgc tttccagtcg ggaaacctgt
     3301 cgtgccagct gcattaacat ggtcatagct gtttccttgc gtattgggcg ctctccgctt
     3361 cctcgctcac tgactcgctg cgctcggtcg ttcgggtaaa gcctggggtg cctaatgagc
     3421 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3481 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3541 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3601 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3661 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3721 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3781 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3841 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3901 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3961 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4021 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4081 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4141 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4201 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4261 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4321 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagaaccacg
     4381 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4441 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4501 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4561 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4621 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4681 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4741 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4801 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4861 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4921 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4981 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5041 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5101 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5161 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5221 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     5281 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     5341 atagggttga gtggccgcta cagggcgctc ccattcgcca ttcaggctgc gcaactgttg
     5401 ggaagggcgt ttcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     5461 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     5521 cggccagtga gcgcgacgta atacgactca ctatagggcg aattggcgga aggccgtcaa
     5581 ggccgtgacg ttaaaactat taagccatcc aatcgaccgt tagtcgaatc aggaccgctg
     5641 gtgcgagaag ccgcgaagta tggcgaatgc atcgtataac gtgtggagtc cgctcattag
     5701 agcgtcatgt ttagacaaga aagctacata tttaattgat cccgatgatt ttattgataa
     5761 attgacccta actccataca cggtattcta caatggcggg gttttggtca aaatttccgg
     5821 actgcgattg tacatgctgt taacggctcc gcccactatt aatgaaatta aaaattccaa
     5881 ttttaaaaaa cgcagcaaga gaaacatttg tatgaaagaa tgcgtagaag gaaagaaaaa
     5941 tgtcgtggac atgctgaaca acaagattaa tatgcctccg tgtataaaaa aaatattgaa
     6001 cgatttgaaa gaaaacaatg taccgcgcgg cggtatgtac aggaagaggt ttatactaaa
     6061 ctgttacatt gcaaacgtgg tttcgtgtgc caagtgtgaa aaccgatgtt taatcaaggc
     6121 tctgacgcat ttctacaacc acgactccaa gtgtgtgggt gaagtcatgc atcttttaat
     6181 caaatcccaa gatgtgtata aaccaccaaa ctgccaaaaa atgaaaactg tggacaagct
     6241 ctgtccgttt gctggcaact gcaagggtct caatcctatt tgtaattatt gaataataaa
     6301 acaattataa atgctaaatt tgttttttat taacgataca aaccaaacgc aacaagaaca
     6361 tttgtagtat tatctataat tgaaaacgcg tagttataat cgctgaggta atatttaaaa
     6421 tcattttcaa atgattcaca gttaatttgc gacaatataa ttttattttc acataaacta
     6481 gacgccttgt cgtcttcttc ttcgtattcc ttctcttttt catttttctc ctcataaaaa
     6541 ttaacatagt tattatcgta tccatatatg tatctatcgt atagagtaaa ttttttgttg
     6601 tcataaatat atatgtcttt tttaatgggg tgtatagtac cgctgcgcat agtttttctg
     6661 taatttacaa cagtgctatt ttctggtagt tcttcggagt gtgttgcttt aattattaaa
     6721 tttatataat caatgaattt gggatcgtcg gttttgtaca atatgttgcc ggcatagtac
     6781 gcagcttctt ctagttcaat tacaccattt tttagcagca ccggattaac ataactttcc
     6841 aaaatgttgt acgaaccgtt aaacaaaaac agttcacctc ccttttctat actattgtct
     6901 gcgagcagtt gtttgttgtt aaaaataaca gccattgtaa tgagacgcac aaactaatat
     6961 cacaagctgc caccattttg ttcgcccagg actctagcta tagttctagt ggttggctac
     7021 gtatactccg gaatattaat agatcatgga gataattaaa atgataacca tctcgcaaat
     7081 aaataagtat tttactgttt tcgtaacagt tttgtaataa aaaaacctat aaat
//
