LOCUS       sequence-224701-        6177 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 6177)
  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 6177)
  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..6177
                     /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             398..703
                     /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     582..601
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             join(765..767,768..770,771..1481)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(765..767,768..770,771..1481)
                     /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(810..831)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1071..1090)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1418..1439
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1491..1508
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1643..1777
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1680..1699)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1734..1753
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1850..2222
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(2574..2591)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2684..3272)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2764..2783)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3443..4234,4235..4303))
                     /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     4066..4085
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4305..4409)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4710..4732
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4724..4741
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4725..4741
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4752..4771
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4752..4770
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5381..5505
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5543..6148)
                     /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 gtgtccgcta tcgtgctgta
      241 cgtgctgctg gctgctgctg ctcactccgc tttcgctgga tccaattggg cggccgcaag
      301 cgctatggcc acaacctgtt acattgcaca agataaaaat atatcatcac gaacagtaaa
      361 actgtctgct tacataaaca gtaatacaag gggtgttatg cagtttaagg tttacaccta
      421 taaaagagag agccgttatc gtctgtttgt ggatgtacag agtgatatta ttgacacgcc
      481 cgggcgacgg atggtgatcc ccctggccag tgcccgtctg ctgtcagata aagtctcccg
      541 tgaactttac ccggtggtgc atatcgggga tgaaagctgg cgcatgatga ccaccgatat
      601 ggccagtgtg ccggtctccg ttatcgggga agaagtggct gatctcagcc accgcgaaaa
      661 tgacatcaaa aacgccatta acctgatgtt ctggggaata taaatgtcag gctcccttat
      721 acacagccag tggcgcgccg gctggctccg ctgctggttc tggcatggtg agcaagggcg
      781 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
      841 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
      901 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
      961 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     1021 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     1081 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     1141 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     1201 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     1261 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     1321 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     1381 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     1441 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gagcagcggc catcaccatc
     1501 accatcacta aagaagcttg tcgagaagta ctagaggatc ataatcagcc ataccacatt
     1561 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     1621 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     1681 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     1741 gtccaaactc atcaatgtat cttatcatgt ctggatctga tcactgcttg agcctaggag
     1801 atccgaacca gataagtgaa agcctgcatc cgcagatctt aattaataaa acacgataca
     1861 ttgttattag tacatttatt aagcgctaga ttctgtgcgt tgttgattta cagacaattg
     1921 ttgtacgtat tttaataatt cattaaattt ataatcttta gggtggtatg ttagagcgaa
     1981 aatcaaatga ttttcagcgt ctttatatct gaatttaaat attaaatcct caatagattt
     2041 gtaaaatagg tttcgattag tttcaaacaa gggttgtttt tccgaaccga tggctggact
     2101 atctaatgga ttttcgctca acgccacaaa acttgccaaa tcttgtagca gcaatctagc
     2161 tttgtcgata ttcgtttgtg ttttgttttg taataaaggt tcgacgtcgt tcaaaatatt
     2221 atgcgctttt gtatttcttt catcactgtc gttagtgtac aattgactcg acgtaaacac
     2281 gttaaataac gcttggacat atttaacatc gggcgtgtta gctttattag gccgattatc
     2341 gtcgtcgtcc caaccctcgt cgttagaagt tgcttccgaa gacgattttg ccatagccac
     2401 acgacgccta ttaattgtgt cggctaacac gtccgcgatc aaatttgtag ttgagctttt
     2461 tgggcctcat gggccttccg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     2521 ctgcattaac atggtcatag ctgtttcctt gcgtattggg cgctctccgc ttcctcgctc
     2581 actgactcgc tgcgctcggt cgttcgggta aagcctgggg tgcctaatga gcaaaaggcc
     2641 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     2701 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     2761 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     2821 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     2881 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     2941 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3001 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     3061 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     3121 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     3181 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     3241 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     3301 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     3361 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     3421 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     3481 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     3541 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     3601 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     3661 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     3721 cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct
     3781 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     3841 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     3901 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     3961 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     4021 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac
     4081 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     4141 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     4201 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     4261 caaaaaaggg aataagggcg acacggaaat gttgaatact cattactctt cctttttcaa
     4321 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     4381 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg
     4441 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     4501 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     4561 tgagtggccg ctacagggcg ctcccattcg ccattcaggc tgcgcaactg ttgggaaggg
     4621 cgtttcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     4681 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     4741 tgagcgcgac gtaatacgac tcactatagg gcgaattggc ggaaggccgt caaggccgtg
     4801 acgttaaaac tattaagcca tccaatcgac cgttagtcga atcaggaccg ctggtgcgag
     4861 aagccgcgaa gtatggcgaa tgcatcgtat aacgtgtgga gtccgctcat tagagcgtca
     4921 tgtttagaca agaaagctac atatttaatt gatcccgatg attttattga taaattgacc
     4981 ctaactccat acacggtatt ctacaatggc ggggttttgg tcaaaatttc cggactgcga
     5041 ttgtacatgc tgttaacggc tccgcccact attaatgaaa ttaaaaattc caattttaaa
     5101 aaacgcagca agagaaacat ttgtatgaaa gaatgcgtag aaggaaagaa aaatgtcgtg
     5161 gacatgctga acaacaagat taatatgcct ccgtgtataa aaaaaatatt gaacgatttg
     5221 aaagaaaaca atgtaccgcg cggcggtatg tacaggaaga ggtttatact aaactgttac
     5281 attgcaaacg tggtttcgtg tgccaagtgt gaaaaccgat gtttaatcaa ggctctgacg
     5341 catttctaca accacgactc caagtgtgtg ggtgaagtca tgcatctttt aatcaaatcc
     5401 caagatgtgt ataaaccacc aaactgccaa aaaatgaaaa ctgtggacaa gctctgtccg
     5461 tttgctggca actgcaaggg tctcaatcct atttgtaatt attgaataat aaaacaatta
     5521 taaatgctaa atttgttttt tattaacgat acaaaccaaa cgcaacaaga acatttgtag
     5581 tattatctat aattgaaaac gcgtagttat aatcgctgag gtaatattta aaatcatttt
     5641 caaatgattc acagttaatt tgcgacaata taattttatt ttcacataaa ctagacgcct
     5701 tgtcgtcttc ttcttcgtat tccttctctt tttcattttt ctcctcataa aaattaacat
     5761 agttattatc gtatccatat atgtatctat cgtatagagt aaattttttg ttgtcataaa
     5821 tatatatgtc ttttttaatg gggtgtatag taccgctgcg catagttttt ctgtaattta
     5881 caacagtgct attttctggt agttcttcgg agtgtgttgc tttaattatt aaatttatat
     5941 aatcaatgaa tttgggatcg tcggttttgt acaatatgtt gccggcatag tacgcagctt
     6001 cttctagttc aattacacca ttttttagca gcaccggatt aacataactt tccaaaatgt
     6061 tgtacgaacc gttaaacaaa aacagttcac ctcccttttc tatactattg tctgcgagca
     6121 gttgtttgtt gttaaaaata acagccattg taatgagacg cacaaactaa tatcaca
//
