LOCUS       sequence-234552-        6182 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with honey bee melitine secretion signal and
            C-terminal eGFP-HIS6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6182)
  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 6182)
  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: pOCC92
FEATURES             Location/Qualifiers
     source          1..6182
                     /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"
     sig_peptide     54..113
                     /codon_start=1
                     /product="signal sequence from honeybee melittin"
                     /label=melittin signal sequence
                     /translation="KFLVNVALVFMVVYISYIYA"
     CDS             234..539
                     /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     418..437
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             join(601..603,604..606,607..1317)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(601..603,604..606,607..1317)
                     /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(646..667)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(907..926)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1254..1275
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1327..1344
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1479..1613
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1516..1535)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1570..1589
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1686..2058
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(2410..2427)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2520..3108)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2600..2619)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3279..4070,4071..4139))
                     /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     3902..3921
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4140..4244)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4545..4567
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4559..4576
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4560..4576
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4587..4606
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4587..4605
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5216..5340
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5378..5983)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     promoter        6091..6182
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     6107..6124
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
ORIGIN
        1 attccggatt attcataccg tcccaccatc gggcgcggat cgccaccatg ggcaaattct
       61 tagtcaacgt tgcccttgtt tttatggtcg tatacatttc ttacatctat gcgggatcca
      121 attgggcggc cgcaagcgct atggccacaa cctgttacat tgcacaagat aaaaatatat
      181 catcacgaac agtaaaactg tctgcttaca taaacagtaa tacaaggggt gttatgcagt
      241 ttaaggttta cacctataaa agagagagcc gttatcgtct gtttgtggat gtacagagtg
      301 atattattga cacgcccggg cgacggatgg tgatccccct ggccagtgcc cgtctgctgt
      361 cagataaagt ctcccgtgaa ctttacccgg tggtgcatat cggggatgaa agctggcgca
      421 tgatgaccac cgatatggcc agtgtgccgg tctccgttat cggggaagaa gtggctgatc
      481 tcagccaccg cgaaaatgac atcaaaaacg ccattaacct gatgttctgg ggaatataaa
      541 tgtcaggctc ccttatacac agccagtggc gcgccggctg gctccgctgc tggttctggc
      601 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
      661 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      721 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
      781 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
      841 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      901 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
      961 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     1021 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     1081 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     1141 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     1201 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     1261 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagagc
     1321 agcggccatc accatcacca tcactaaaga agcttgtcga gaagtactag aggatcataa
     1381 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     1441 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     1501 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     1561 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atctgatcac
     1621 tgcttgagcc taggagatcc gaaccagata agtgaaagcc tgcatccgca gatcttaatt
     1681 aataaaacac gatacattgt tattagtaca tttattaagc gctagattct gtgcgttgtt
     1741 gatttacaga caattgttgt acgtatttta ataattcatt aaatttataa tctttagggt
     1801 ggtatgttag agcgaaaatc aaatgatttt cagcgtcttt atatctgaat ttaaatatta
     1861 aatcctcaat agatttgtaa aataggtttc gattagtttc aaacaagggt tgtttttccg
     1921 aaccgatggc tggactatct aatggatttt cgctcaacgc cacaaaactt gccaaatctt
     1981 gtagcagcaa tctagctttg tcgatattcg tttgtgtttt gttttgtaat aaaggttcga
     2041 cgtcgttcaa aatattatgc gcttttgtat ttctttcatc actgtcgtta gtgtacaatt
     2101 gactcgacgt aaacacgtta aataacgctt ggacatattt aacatcgggc gtgttagctt
     2161 tattaggccg attatcgtcg tcgtcccaac cctcgtcgtt agaagttgct tccgaagacg
     2221 attttgccat agccacacga cgcctattaa ttgtgtcggc taacacgtcc gcgatcaaat
     2281 ttgtagttga gctttttggg cctcatgggc cttccgctca ctgcccgctt tccagtcggg
     2341 aaacctgtcg tgccagctgc attaacatgg tcatagctgt ttccttgcgt attgggcgct
     2401 ctccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cgggtaaagc ctggggtgcc
     2461 taatgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2521 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2581 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     2641 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2701 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2761 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2821 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2881 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     2941 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3001 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3061 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3121 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3181 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3241 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3301 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     3361 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     3421 gaaccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     3481 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     3541 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     3601 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     3661 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     3721 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     3781 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     3841 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     3901 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     3961 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4021 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     4081 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4141 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4201 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     4261 gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     4321 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     4381 agaccgagat agggttgagt ggccgctaca gggcgctccc attcgccatt caggctgcgc
     4441 aactgttggg aagggcgttt cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     4501 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     4561 taaaacgacg gccagtgagc gcgacgtaat acgactcact atagggcgaa ttggcggaag
     4621 gccgtcaagg ccgtgacgtt aaaactatta agccatccaa tcgaccgtta gtcgaatcag
     4681 gaccgctggt gcgagaagcc gcgaagtatg gcgaatgcat cgtataacgt gtggagtccg
     4741 ctcattagag cgtcatgttt agacaagaaa gctacatatt taattgatcc cgatgatttt
     4801 attgataaat tgaccctaac tccatacacg gtattctaca atggcggggt tttggtcaaa
     4861 atttccggac tgcgattgta catgctgtta acggctccgc ccactattaa tgaaattaaa
     4921 aattccaatt ttaaaaaacg cagcaagaga aacatttgta tgaaagaatg cgtagaagga
     4981 aagaaaaatg tcgtggacat gctgaacaac aagattaata tgcctccgtg tataaaaaaa
     5041 atattgaacg atttgaaaga aaacaatgta ccgcgcggcg gtatgtacag gaagaggttt
     5101 atactaaact gttacattgc aaacgtggtt tcgtgtgcca agtgtgaaaa ccgatgttta
     5161 atcaaggctc tgacgcattt ctacaaccac gactccaagt gtgtgggtga agtcatgcat
     5221 cttttaatca aatcccaaga tgtgtataaa ccaccaaact gccaaaaaat gaaaactgtg
     5281 gacaagctct gtccgtttgc tggcaactgc aagggtctca atcctatttg taattattga
     5341 ataataaaac aattataaat gctaaatttg ttttttatta acgatacaaa ccaaacgcaa
     5401 caagaacatt tgtagtatta tctataattg aaaacgcgta gttataatcg ctgaggtaat
     5461 atttaaaatc attttcaaat gattcacagt taatttgcga caatataatt ttattttcac
     5521 ataaactaga cgccttgtcg tcttcttctt cgtattcctt ctctttttca tttttctcct
     5581 cataaaaatt aacatagtta ttatcgtatc catatatgta tctatcgtat agagtaaatt
     5641 ttttgttgtc ataaatatat atgtcttttt taatggggtg tatagtaccg ctgcgcatag
     5701 tttttctgta atttacaaca gtgctatttt ctggtagttc ttcggagtgt gttgctttaa
     5761 ttattaaatt tatataatca atgaatttgg gatcgtcggt tttgtacaat atgttgccgg
     5821 catagtacgc agcttcttct agttcaatta caccattttt tagcagcacc ggattaacat
     5881 aactttccaa aatgttgtac gaaccgttaa acaaaaacag ttcacctccc ttttctatac
     5941 tattgtctgc gagcagttgt ttgttgttaa aaataacagc cattgtaatg agacgcacaa
     6001 actaatatca caagctgcca ccattttgtt cgcccaggac tctagctata gttctagtgg
     6061 ttggctacgt atactccgga atattaatag atcatggaga taattaaaat gataaccatc
     6121 tcgcaaataa ataagtattt tactgttttc gtaacagttt tgtaataaaa aaacctataa
     6181 at
//
