LOCUS       sequence-224690-        6064 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with N-terminal HIS6, cleavable with 3C , 
            and N-terminal GST.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6064)
  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 6064)
  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: pOCC81
FEATURES             Location/Qualifiers
     source          1..6064
                     /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             230..247
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             260..283
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             284..937
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     895..917
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1052..1357
                     /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     1236..1255
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    1530..1664
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1567..1586)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1621..1640
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1737..2109
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(2461..2478)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2571..3159)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2651..2670)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3330..4121,4122..4190))
                     /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     3953..3972
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4192..4296)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4597..4619
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4611..4628
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4612..4628
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4639..4658
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4639..4657
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5268..5392
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5430..6035)
                     /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 ggcagcagcc atcaccatca
      241 ccatcacagc agcggcagac tggaagttct gttccagggg ccgatgtccc ctatactagg
      301 ttattggaaa attaagggcc ttgtgcaacc cactcgactt cttttggaat atcttgaaga
      361 aaaatatgaa gagcatttgt atgagcgcga tgaaggtgat aaatggcgaa acaaaaagtt
      421 tgaattgggt ttggagtttc ccaatcttcc ttattatatt gatggtgatg ttaaattaac
      481 acagtctatg gccatcatac gttatatagc tgacaagcac aacatgttgg gtggttgtcc
      541 aaaagagcgt gcagagattt caatgcttga aggagcggtt ttggatatta gatacggtgt
      601 ttcgagaatt gcatatagta aagactttga aactctcaaa gttgattttc ttagcaagct
      661 acctgaaatg ctgaaaatgt tcgaagatcg tttatgtcat aaaacatatt taaatggtga
      721 tcatgtaacc catcctgact tcatgttgta tgacgctctt gatgttgttt tatacatgga
      781 cccaatgtgc ctggatgcgt tcccaaaatt agtttgtttt aaaaaacgta ttgaagctat
      841 cccacaaatt gataagtact tgaaatccag caagtatata gcatggcctt tgcagggctg
      901 gcaagccacg tttggtggtg gcgaccatcc tccaaaatcg gatgcggccg caagcgctat
      961 ggccacaacc tgttacattg cacaagataa aaatatatca tcacgaacag taaaactgtc
     1021 tgcttacata aacagtaata caaggggtgt tatgcagttt aaggtttaca cctataaaag
     1081 agagagccgt tatcgtctgt ttgtggatgt acagagtgat attattgaca cgcccgggcg
     1141 acggatggtg atccccctgg ccagtgcccg tctgctgtca gataaagtct cccgtgaact
     1201 ttacccggtg gtgcatatcg gggatgaaag ctggcgcatg atgaccaccg atatggccag
     1261 tgtgccggtc tccgttatcg gggaagaagt ggctgatctc agccaccgcg aaaatgacat
     1321 caaaaacgcc attaacctga tgttctgggg aatataaatg tcaggctccc ttatacacag
     1381 ccagtggcgc gccgtaatag aagcttgtcg agaagtacta gaggatcata atcagccata
     1441 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     1501 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     1561 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     1621 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatctgatca ctgcttgagc
     1681 ctaggagatc cgaaccagat aagtgaaagc ctgcatccgc agatcttaat taataaaaca
     1741 cgatacattg ttattagtac atttattaag cgctagattc tgtgcgttgt tgatttacag
     1801 acaattgttg tacgtatttt aataattcat taaatttata atctttaggg tggtatgtta
     1861 gagcgaaaat caaatgattt tcagcgtctt tatatctgaa tttaaatatt aaatcctcaa
     1921 tagatttgta aaataggttt cgattagttt caaacaaggg ttgtttttcc gaaccgatgg
     1981 ctggactatc taatggattt tcgctcaacg ccacaaaact tgccaaatct tgtagcagca
     2041 atctagcttt gtcgatattc gtttgtgttt tgttttgtaa taaaggttcg acgtcgttca
     2101 aaatattatg cgcttttgta tttctttcat cactgtcgtt agtgtacaat tgactcgacg
     2161 taaacacgtt aaataacgct tggacatatt taacatcggg cgtgttagct ttattaggcc
     2221 gattatcgtc gtcgtcccaa ccctcgtcgt tagaagttgc ttccgaagac gattttgcca
     2281 tagccacacg acgcctatta attgtgtcgg ctaacacgtc cgcgatcaaa tttgtagttg
     2341 agctttttgg gcctcatggg ccttccgctc actgcccgct ttccagtcgg gaaacctgtc
     2401 gtgccagctg cattaacatg gtcatagctg tttccttgcg tattgggcgc tctccgcttc
     2461 ctcgctcact gactcgctgc gctcggtcgt tcgggtaaag cctggggtgc ctaatgagca
     2521 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     2581 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2641 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2701 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2761 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2821 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2881 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2941 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3001 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3061 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3121 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3181 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     3241 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     3301 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     3361 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     3421 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     3481 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     3541 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     3601 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg
     3661 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     3721 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     3781 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     3841 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     3901 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc
     3961 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     4021 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     4081 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     4141 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat tactcttcct
     4201 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4261 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4321 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     4381 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     4441 atagggttga gtggccgcta cagggcgctc ccattcgcca ttcaggctgc gcaactgttg
     4501 ggaagggcgt ttcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     4561 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     4621 cggccagtga gcgcgacgta atacgactca ctatagggcg aattggcgga aggccgtcaa
     4681 ggccgtgacg ttaaaactat taagccatcc aatcgaccgt tagtcgaatc aggaccgctg
     4741 gtgcgagaag ccgcgaagta tggcgaatgc atcgtataac gtgtggagtc cgctcattag
     4801 agcgtcatgt ttagacaaga aagctacata tttaattgat cccgatgatt ttattgataa
     4861 attgacccta actccataca cggtattcta caatggcggg gttttggtca aaatttccgg
     4921 actgcgattg tacatgctgt taacggctcc gcccactatt aatgaaatta aaaattccaa
     4981 ttttaaaaaa cgcagcaaga gaaacatttg tatgaaagaa tgcgtagaag gaaagaaaaa
     5041 tgtcgtggac atgctgaaca acaagattaa tatgcctccg tgtataaaaa aaatattgaa
     5101 cgatttgaaa gaaaacaatg taccgcgcgg cggtatgtac aggaagaggt ttatactaaa
     5161 ctgttacatt gcaaacgtgg tttcgtgtgc caagtgtgaa aaccgatgtt taatcaaggc
     5221 tctgacgcat ttctacaacc acgactccaa gtgtgtgggt gaagtcatgc atcttttaat
     5281 caaatcccaa gatgtgtata aaccaccaaa ctgccaaaaa atgaaaactg tggacaagct
     5341 ctgtccgttt gctggcaact gcaagggtct caatcctatt tgtaattatt gaataataaa
     5401 acaattataa atgctaaatt tgttttttat taacgataca aaccaaacgc aacaagaaca
     5461 tttgtagtat tatctataat tgaaaacgcg tagttataat cgctgaggta atatttaaaa
     5521 tcattttcaa atgattcaca gttaatttgc gacaatataa ttttattttc acataaacta
     5581 gacgccttgt cgtcttcttc ttcgtattcc ttctcttttt catttttctc ctcataaaaa
     5641 ttaacatagt tattatcgta tccatatatg tatctatcgt atagagtaaa ttttttgttg
     5701 tcataaatat atatgtcttt tttaatgggg tgtatagtac cgctgcgcat agtttttctg
     5761 taatttacaa cagtgctatt ttctggtagt tcttcggagt gtgttgcttt aattattaaa
     5821 tttatataat caatgaattt gggatcgtcg gttttgtaca atatgttgcc ggcatagtac
     5881 gcagcttctt ctagttcaat tacaccattt tttagcagca ccggattaac ataactttcc
     5941 aaaatgttgt acgaaccgtt aaacaaaaac agttcacctc ccttttctat actattgtct
     6001 gcgagcagtt gtttgttgtt aaaaataaca gccattgtaa tgagacgcac aaactaatat
     6061 caca
//
