LOCUS       sequence-232347-        6176 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with GP64 viral secretion signal and 
            C-terminal eGFP-HIS6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6176)
  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 6176)
  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: pOCC93
FEATURES             Location/Qualifiers
     source          1..6176
                     /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             228..533
                     /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     412..431
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             join(595..597,598..600,601..1311)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(595..597,598..600,601..1311)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(640..661)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(901..920)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1248..1269
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1321..1338
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1473..1607
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1510..1529)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1564..1583
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1680..2052
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(2404..2421)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2514..3102)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2594..2613)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3273..4064,4065..4133))
                     /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     3896..3915
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4134..4238)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4539..4561
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4553..4570
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4554..4570
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4581..4600
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4581..4599
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5210..5334
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5372..5977)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     promoter        6085..6176
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     6101..6118
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
ORIGIN
        1 attccggatt attcataccg tcccaccatc gggcgcggat cgccaccatg gtgtccgcta
       61 tcgtgctgta cgtgctgctg gctgctgctg ctcactccgc tttcgctgga tccaattggg
      121 cggccgcaag cgctatggcc acaacctgtt acattgcaca agataaaaat atatcatcac
      181 gaacagtaaa actgtctgct tacataaaca gtaatacaag gggtgttatg cagtttaagg
      241 tttacaccta taaaagagag agccgttatc gtctgtttgt ggatgtacag agtgatatta
      301 ttgacacgcc cgggcgacgg atggtgatcc ccctggccag tgcccgtctg ctgtcagata
      361 aagtctcccg tgaactttac ccggtggtgc atatcgggga tgaaagctgg cgcatgatga
      421 ccaccgatat ggccagtgtg ccggtctccg ttatcgggga agaagtggct gatctcagcc
      481 accgcgaaaa tgacatcaaa aacgccatta acctgatgtt ctggggaata taaatgtcag
      541 gctcccttat acacagccag tggcgcgccg gctggctccg ctgctggttc tggcatggtg
      601 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
      661 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
      721 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
      781 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
      841 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
      901 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
      961 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     1021 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     1081 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     1141 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     1201 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     1261 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gagcagcggc
     1321 catcaccatc accatcacta aagaagcttg tcgagaagta ctagaggatc ataatcagcc
     1381 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     1441 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     1501 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     1561 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatctga tcactgcttg
     1621 agcctaggag atccgaacca gataagtgaa agcctgcatc cgcagatctt aattaataaa
     1681 acacgataca ttgttattag tacatttatt aagcgctaga ttctgtgcgt tgttgattta
     1741 cagacaattg ttgtacgtat tttaataatt cattaaattt ataatcttta gggtggtatg
     1801 ttagagcgaa aatcaaatga ttttcagcgt ctttatatct gaatttaaat attaaatcct
     1861 caatagattt gtaaaatagg tttcgattag tttcaaacaa gggttgtttt tccgaaccga
     1921 tggctggact atctaatgga ttttcgctca acgccacaaa acttgccaaa tcttgtagca
     1981 gcaatctagc tttgtcgata ttcgtttgtg ttttgttttg taataaaggt tcgacgtcgt
     2041 tcaaaatatt atgcgctttt gtatttcttt catcactgtc gttagtgtac aattgactcg
     2101 acgtaaacac gttaaataac gcttggacat atttaacatc gggcgtgtta gctttattag
     2161 gccgattatc gtcgtcgtcc caaccctcgt cgttagaagt tgcttccgaa gacgattttg
     2221 ccatagccac acgacgccta ttaattgtgt cggctaacac gtccgcgatc aaatttgtag
     2281 ttgagctttt tgggcctcat gggccttccg ctcactgccc gctttccagt cgggaaacct
     2341 gtcgtgccag ctgcattaac atggtcatag ctgtttcctt gcgtattggg cgctctccgc
     2401 ttcctcgctc actgactcgc tgcgctcggt cgttcgggta aagcctgggg tgcctaatga
     2461 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2521 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2581 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2641 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2701 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2761 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2821 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2881 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2941 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3001 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3061 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3121 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     3181 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     3241 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     3301 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     3361 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaacca
     3421 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     3481 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     3541 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     3601 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3661 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     3721 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     3781 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     3841 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     3901 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     3961 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     4021 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     4081 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     4141 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     4201 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     4261 cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     4321 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     4381 agatagggtt gagtggccgc tacagggcgc tcccattcgc cattcaggct gcgcaactgt
     4441 tgggaagggc gtttcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     4501 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     4561 gacggccagt gagcgcgacg taatacgact cactataggg cgaattggcg gaaggccgtc
     4621 aaggccgtga cgttaaaact attaagccat ccaatcgacc gttagtcgaa tcaggaccgc
     4681 tggtgcgaga agccgcgaag tatggcgaat gcatcgtata acgtgtggag tccgctcatt
     4741 agagcgtcat gtttagacaa gaaagctaca tatttaattg atcccgatga ttttattgat
     4801 aaattgaccc taactccata cacggtattc tacaatggcg gggttttggt caaaatttcc
     4861 ggactgcgat tgtacatgct gttaacggct ccgcccacta ttaatgaaat taaaaattcc
     4921 aattttaaaa aacgcagcaa gagaaacatt tgtatgaaag aatgcgtaga aggaaagaaa
     4981 aatgtcgtgg acatgctgaa caacaagatt aatatgcctc cgtgtataaa aaaaatattg
     5041 aacgatttga aagaaaacaa tgtaccgcgc ggcggtatgt acaggaagag gtttatacta
     5101 aactgttaca ttgcaaacgt ggtttcgtgt gccaagtgtg aaaaccgatg tttaatcaag
     5161 gctctgacgc atttctacaa ccacgactcc aagtgtgtgg gtgaagtcat gcatctttta
     5221 atcaaatccc aagatgtgta taaaccacca aactgccaaa aaatgaaaac tgtggacaag
     5281 ctctgtccgt ttgctggcaa ctgcaagggt ctcaatccta tttgtaatta ttgaataata
     5341 aaacaattat aaatgctaaa tttgtttttt attaacgata caaaccaaac gcaacaagaa
     5401 catttgtagt attatctata attgaaaacg cgtagttata atcgctgagg taatatttaa
     5461 aatcattttc aaatgattca cagttaattt gcgacaatat aattttattt tcacataaac
     5521 tagacgcctt gtcgtcttct tcttcgtatt ccttctcttt ttcatttttc tcctcataaa
     5581 aattaacata gttattatcg tatccatata tgtatctatc gtatagagta aattttttgt
     5641 tgtcataaat atatatgtct tttttaatgg ggtgtatagt accgctgcgc atagtttttc
     5701 tgtaatttac aacagtgcta ttttctggta gttcttcgga gtgtgttgct ttaattatta
     5761 aatttatata atcaatgaat ttgggatcgt cggttttgta caatatgttg ccggcatagt
     5821 acgcagcttc ttctagttca attacaccat tttttagcag caccggatta acataacttt
     5881 ccaaaatgtt gtacgaaccg ttaaacaaaa acagttcacc tcccttttct atactattgt
     5941 ctgcgagcag ttgtttgttg ttaaaaataa cagccattgt aatgagacgc acaaactaat
     6001 atcacaagct gccaccattt tgttcgccca ggactctagc tatagttcta gtggttggct
     6061 acgtatactc cggaatatta atagatcatg gagataatta aaatgataac catctcgcaa
     6121 ataaataagt attttactgt tttcgtaaca gttttgtaat aaaaaaacct ataaat
//
