LOCUS       sequence-224653-        5404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with N-terminal HIS6 tag, cleavable with 3C 
            protease.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5404)
  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 5404)
  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: pOCC5
FEATURES             Location/Qualifiers
     source          1..5404
                     /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             392..697
                     /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     576..595
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    870..1004
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(907..926)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     961..980
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1077..1449
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(1801..1818)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1911..2499)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1991..2010)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2670..3461,3462..3530))
                     /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     3293..3312
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3532..3636)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3937..3959
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3951..3968
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3952..3968
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3979..3998
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3979..3997
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4608..4732
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(4770..5375)
                     /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 ccggcggccg caagcgctat
      301 ggccacaacc tgttacattg cacaagataa aaatatatca tcacgaacag taaaactgtc
      361 tgcttacata aacagtaata caaggggtgt tatgcagttt aaggtttaca cctataaaag
      421 agagagccgt tatcgtctgt ttgtggatgt acagagtgat attattgaca cgcccgggcg
      481 acggatggtg atccccctgg ccagtgcccg tctgctgtca gataaagtct cccgtgaact
      541 ttacccggtg gtgcatatcg gggatgaaag ctggcgcatg atgaccaccg atatggccag
      601 tgtgccggtc tccgttatcg gggaagaagt ggctgatctc agccaccgcg aaaatgacat
      661 caaaaacgcc attaacctga tgttctgggg aatataaatg tcaggctccc ttatacacag
      721 ccagtggcgc gccgtaatag aagcttgtcg agaagtacta gaggatcata atcagccata
      781 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
      841 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
      901 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
      961 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatctgatca ctgcttgagc
     1021 ctaggagatc cgaaccagat aagtgaaagc ctgcatccgc agatcttaat taataaaaca
     1081 cgatacattg ttattagtac atttattaag cgctagattc tgtgcgttgt tgatttacag
     1141 acaattgttg tacgtatttt aataattcat taaatttata atctttaggg tggtatgtta
     1201 gagcgaaaat caaatgattt tcagcgtctt tatatctgaa tttaaatatt aaatcctcaa
     1261 tagatttgta aaataggttt cgattagttt caaacaaggg ttgtttttcc gaaccgatgg
     1321 ctggactatc taatggattt tcgctcaacg ccacaaaact tgccaaatct tgtagcagca
     1381 atctagcttt gtcgatattc gtttgtgttt tgttttgtaa taaaggttcg acgtcgttca
     1441 aaatattatg cgcttttgta tttctttcat cactgtcgtt agtgtacaat tgactcgacg
     1501 taaacacgtt aaataacgct tggacatatt taacatcggg cgtgttagct ttattaggcc
     1561 gattatcgtc gtcgtcccaa ccctcgtcgt tagaagttgc ttccgaagac gattttgcca
     1621 tagccacacg acgcctatta attgtgtcgg ctaacacgtc cgcgatcaaa tttgtagttg
     1681 agctttttgg gcctcatggg ccttccgctc actgcccgct ttccagtcgg gaaacctgtc
     1741 gtgccagctg cattaacatg gtcatagctg tttccttgcg tattgggcgc tctccgcttc
     1801 ctcgctcact gactcgctgc gctcggtcgt tcgggtaaag cctggggtgc ctaatgagca
     1861 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     1921 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     1981 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2041 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2101 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2161 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2221 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2281 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     2341 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     2401 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     2461 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     2521 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     2581 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     2641 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     2701 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     2761 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     2821 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     2881 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     2941 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg
     3001 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     3061 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     3121 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     3181 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     3241 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc
     3301 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     3361 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     3421 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     3481 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat tactcttcct
     3541 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     3601 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     3661 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     3721 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     3781 atagggttga gtggccgcta cagggcgctc ccattcgcca ttcaggctgc gcaactgttg
     3841 ggaagggcgt ttcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     3901 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     3961 cggccagtga gcgcgacgta atacgactca ctatagggcg aattggcgga aggccgtcaa
     4021 ggccgtgacg ttaaaactat taagccatcc aatcgaccgt tagtcgaatc aggaccgctg
     4081 gtgcgagaag ccgcgaagta tggcgaatgc atcgtataac gtgtggagtc cgctcattag
     4141 agcgtcatgt ttagacaaga aagctacata tttaattgat cccgatgatt ttattgataa
     4201 attgacccta actccataca cggtattcta caatggcggg gttttggtca aaatttccgg
     4261 actgcgattg tacatgctgt taacggctcc gcccactatt aatgaaatta aaaattccaa
     4321 ttttaaaaaa cgcagcaaga gaaacatttg tatgaaagaa tgcgtagaag gaaagaaaaa
     4381 tgtcgtggac atgctgaaca acaagattaa tatgcctccg tgtataaaaa aaatattgaa
     4441 cgatttgaaa gaaaacaatg taccgcgcgg cggtatgtac aggaagaggt ttatactaaa
     4501 ctgttacatt gcaaacgtgg tttcgtgtgc caagtgtgaa aaccgatgtt taatcaaggc
     4561 tctgacgcat ttctacaacc acgactccaa gtgtgtgggt gaagtcatgc atcttttaat
     4621 caaatcccaa gatgtgtata aaccaccaaa ctgccaaaaa atgaaaactg tggacaagct
     4681 ctgtccgttt gctggcaact gcaagggtct caatcctatt tgtaattatt gaataataaa
     4741 acaattataa atgctaaatt tgttttttat taacgataca aaccaaacgc aacaagaaca
     4801 tttgtagtat tatctataat tgaaaacgcg tagttataat cgctgaggta atatttaaaa
     4861 tcattttcaa atgattcaca gttaatttgc gacaatataa ttttattttc acataaacta
     4921 gacgccttgt cgtcttcttc ttcgtattcc ttctcttttt catttttctc ctcataaaaa
     4981 ttaacatagt tattatcgta tccatatatg tatctatcgt atagagtaaa ttttttgttg
     5041 tcataaatat atatgtcttt tttaatgggg tgtatagtac cgctgcgcat agtttttctg
     5101 taatttacaa cagtgctatt ttctggtagt tcttcggagt gtgttgcttt aattattaaa
     5161 tttatataat caatgaattt gggatcgtcg gttttgtaca atatgttgcc ggcatagtac
     5221 gcagcttctt ctagttcaat tacaccattt tttagcagca ccggattaac ataactttcc
     5281 aaaatgttgt acgaaccgtt aaacaaaaac agttcacctc ccttttctat actattgtct
     5341 gcgagcagtt gtttgttgtt aaaaataaca gccattgtaa tgagacgcac aaactaatat
     5401 caca
//
