LOCUS       sequence-224668-        6154 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with C-terminal monomeric GFP, cleavable 
            with TEV, and C-terminal HIS6 tag, cleavable with 3C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6154)
  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 6154)
  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: pOCC70
FEATURES             Location/Qualifiers
     source          1..6154
                     /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"
     regulatory      216..225
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             329..634
                     /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     513..532
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             690..710
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(711..713,714..716,717..1427)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(756..777)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1017..1036)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1364..1385
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1428..1451
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1461..1478
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1620..1754
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1657..1676)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1711..1730
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1827..2199
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(2551..2568)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2661..3249)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2741..2760)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3420..4211,4212..4280))
                     /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     4043..4062
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4282..4386)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4687..4709
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4701..4718
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4702..4718
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4729..4748
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4729..4747
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5358..5482
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5520..6125)
                     /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 gcggccgcaa gcgctatggc
      241 cacaacctgt tacattgcac aagataaaaa tatatcatca cgaacagtaa aactgtctgc
      301 ttacataaac agtaatacaa ggggtgttat gcagtttaag gtttacacct ataaaagaga
      361 gagccgttat cgtctgtttg tggatgtaca gagtgatatt attgacacgc ccgggcgacg
      421 gatggtgatc cccctggcca gtgcccgtct gctgtcagat aaagtctccc gtgaacttta
      481 cccggtggtg catatcgggg atgaaagctg gcgcatgatg accaccgata tggccagtgt
      541 gccggtctcc gttatcgggg aagaagtggc tgatctcagc caccgcgaaa atgacatcaa
      601 aaacgccatt aacctgatgt tctggggaat ataaatgtca ggctccctta tacacagcca
      661 gtggcgcgcc gggatccagc agcggcagag aaaacttgta tttccagggc atggtgagca
      721 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
      781 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
      841 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
      901 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
      961 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     1021 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     1081 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     1141 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     1201 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     1261 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     1321 cccagtccaa actgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     1381 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagctg gaagttctgt
     1441 tccaggggcc gggcagcagc catcaccatc accatcacag tggataatag aagcttgtcg
     1501 agaagtacta gaggatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     1561 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     1621 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     1681 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     1741 atcatgtctg gatctgatca ctgcttgagc ctaggagatc cgaaccagat aagtgaaagc
     1801 ctgcatccgc agatcttaat taataaaaca cgatacattg ttattagtac atttattaag
     1861 cgctagattc tgtgcgttgt tgatttacag acaattgttg tacgtatttt aataattcat
     1921 taaatttata atctttaggg tggtatgtta gagcgaaaat caaatgattt tcagcgtctt
     1981 tatatctgaa tttaaatatt aaatcctcaa tagatttgta aaataggttt cgattagttt
     2041 caaacaaggg ttgtttttcc gaaccgatgg ctggactatc taatggattt tcgctcaacg
     2101 ccacaaaact tgccaaatct tgtagcagca atctagcttt gtcgatattc gtttgtgttt
     2161 tgttttgtaa taaaggttcg acgtcgttca aaatattatg cgcttttgta tttctttcat
     2221 cactgtcgtt agtgtacaat tgactcgacg taaacacgtt aaataacgct tggacatatt
     2281 taacatcggg cgtgttagct ttattaggcc gattatcgtc gtcgtcccaa ccctcgtcgt
     2341 tagaagttgc ttccgaagac gattttgcca tagccacacg acgcctatta attgtgtcgg
     2401 ctaacacgtc cgcgatcaaa tttgtagttg agctttttgg gcctcatggg ccttccgctc
     2461 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaacatg gtcatagctg
     2521 tttccttgcg tattgggcgc tctccgcttc ctcgctcact gactcgctgc gctcggtcgt
     2581 tcgggtaaag cctggggtgc ctaatgagca aaaggccagc aaaaggccag gaaccgtaaa
     2641 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2701 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2761 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2821 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     2881 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     2941 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3001 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3061 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3121 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3181 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3241 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3301 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3361 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3421 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3481 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3541 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     3601 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3661 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     3721 gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     3781 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     3841 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     3901 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     3961 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     4021 tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     4081 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4141 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4201 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4261 cggaaatgtt gaatactcat tactcttcct ttttcaatat tattgaagca tttatcaggg
     4321 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4381 tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa
     4441 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     4501 atcccttata aatcaaaaga atagaccgag atagggttga gtggccgcta cagggcgctc
     4561 ccattcgcca ttcaggctgc gcaactgttg ggaagggcgt ttcggtgcgg gcctcttcgc
     4621 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     4681 ggttttccca gtcacgacgt tgtaaaacga cggccagtga gcgcgacgta atacgactca
     4741 ctatagggcg aattggcgga aggccgtcaa ggccgtgacg ttaaaactat taagccatcc
     4801 aatcgaccgt tagtcgaatc aggaccgctg gtgcgagaag ccgcgaagta tggcgaatgc
     4861 atcgtataac gtgtggagtc cgctcattag agcgtcatgt ttagacaaga aagctacata
     4921 tttaattgat cccgatgatt ttattgataa attgacccta actccataca cggtattcta
     4981 caatggcggg gttttggtca aaatttccgg actgcgattg tacatgctgt taacggctcc
     5041 gcccactatt aatgaaatta aaaattccaa ttttaaaaaa cgcagcaaga gaaacatttg
     5101 tatgaaagaa tgcgtagaag gaaagaaaaa tgtcgtggac atgctgaaca acaagattaa
     5161 tatgcctccg tgtataaaaa aaatattgaa cgatttgaaa gaaaacaatg taccgcgcgg
     5221 cggtatgtac aggaagaggt ttatactaaa ctgttacatt gcaaacgtgg tttcgtgtgc
     5281 caagtgtgaa aaccgatgtt taatcaaggc tctgacgcat ttctacaacc acgactccaa
     5341 gtgtgtgggt gaagtcatgc atcttttaat caaatcccaa gatgtgtata aaccaccaaa
     5401 ctgccaaaaa atgaaaactg tggacaagct ctgtccgttt gctggcaact gcaagggtct
     5461 caatcctatt tgtaattatt gaataataaa acaattataa atgctaaatt tgttttttat
     5521 taacgataca aaccaaacgc aacaagaaca tttgtagtat tatctataat tgaaaacgcg
     5581 tagttataat cgctgaggta atatttaaaa tcattttcaa atgattcaca gttaatttgc
     5641 gacaatataa ttttattttc acataaacta gacgccttgt cgtcttcttc ttcgtattcc
     5701 ttctcttttt catttttctc ctcataaaaa ttaacatagt tattatcgta tccatatatg
     5761 tatctatcgt atagagtaaa ttttttgttg tcataaatat atatgtcttt tttaatgggg
     5821 tgtatagtac cgctgcgcat agtttttctg taatttacaa cagtgctatt ttctggtagt
     5881 tcttcggagt gtgttgcttt aattattaaa tttatataat caatgaattt gggatcgtcg
     5941 gttttgtaca atatgttgcc ggcatagtac gcagcttctt ctagttcaat tacaccattt
     6001 tttagcagca ccggattaac ataactttcc aaaatgttgt acgaaccgtt aaacaaaaac
     6061 agttcacctc ccttttctat actattgtct gcgagcagtt gtttgttgtt aaaaataaca
     6121 gccattgtaa tgagacgcac aaactaatat caca
//
