LOCUS       sequence-224699-        5473 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 HIS6, cleavable with 3C protease.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5473)
  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 5473)
  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: pOCC167
FEATURES             Location/Qualifiers
     source          1..5473
                     /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"
     sig_peptide     224..283
                     /codon_start=1
                     /product="signal sequence from honeybee melittin"
                     /label=melittin signal sequence
                     /translation="KFLVNVALVFMVVYISYIYA"
     CDS             404..709
                     /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     588..607
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             747..770
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             780..797
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    939..1073
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(976..995)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1030..1049
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1146..1518
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(1870..1887)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1980..2568)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2060..2079)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2739..3530,3531..3599))
                     /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     3362..3381
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3601..3705)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4006..4028
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4020..4037
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4021..4037
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4048..4067
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4048..4066
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4677..4801
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(4839..5444)
                     /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 ggcaaattct tagtcaacgt
      241 tgcccttgtt tttatggtcg tatacatttc ttacatctat gcgggatcca attgggcggc
      301 cgcaagcgct atggccacaa cctgttacat tgcacaagat aaaaatatat catcacgaac
      361 agtaaaactg tctgcttaca taaacagtaa tacaaggggt gttatgcagt ttaaggttta
      421 cacctataaa agagagagcc gttatcgtct gtttgtggat gtacagagtg atattattga
      481 cacgcccggg cgacggatgg tgatccccct ggccagtgcc cgtctgctgt cagataaagt
      541 ctcccgtgaa ctttacccgg tggtgcatat cggggatgaa agctggcgca tgatgaccac
      601 cgatatggcc agtgtgccgg tctccgttat cggggaagaa gtggctgatc tcagccaccg
      661 cgaaaatgac atcaaaaacg ccattaacct gatgttctgg ggaatataaa tgtcaggctc
      721 ccttatacac agccagtggc gcgccgctgg aagttctgtt ccaggggccg ggcagcagcc
      781 atcaccatca ccatcacagt ggataataga agcttgtcga gaagtactag aggatcataa
      841 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
      901 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
      961 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     1021 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atctgatcac
     1081 tgcttgagcc taggagatcc gaaccagata agtgaaagcc tgcatccgca gatcttaatt
     1141 aataaaacac gatacattgt tattagtaca tttattaagc gctagattct gtgcgttgtt
     1201 gatttacaga caattgttgt acgtatttta ataattcatt aaatttataa tctttagggt
     1261 ggtatgttag agcgaaaatc aaatgatttt cagcgtcttt atatctgaat ttaaatatta
     1321 aatcctcaat agatttgtaa aataggtttc gattagtttc aaacaagggt tgtttttccg
     1381 aaccgatggc tggactatct aatggatttt cgctcaacgc cacaaaactt gccaaatctt
     1441 gtagcagcaa tctagctttg tcgatattcg tttgtgtttt gttttgtaat aaaggttcga
     1501 cgtcgttcaa aatattatgc gcttttgtat ttctttcatc actgtcgtta gtgtacaatt
     1561 gactcgacgt aaacacgtta aataacgctt ggacatattt aacatcgggc gtgttagctt
     1621 tattaggccg attatcgtcg tcgtcccaac cctcgtcgtt agaagttgct tccgaagacg
     1681 attttgccat agccacacga cgcctattaa ttgtgtcggc taacacgtcc gcgatcaaat
     1741 ttgtagttga gctttttggg cctcatgggc cttccgctca ctgcccgctt tccagtcggg
     1801 aaacctgtcg tgccagctgc attaacatgg tcatagctgt ttccttgcgt attgggcgct
     1861 ctccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cgggtaaagc ctggggtgcc
     1921 taatgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     1981 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2041 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     2101 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2161 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2221 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2281 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2341 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     2401 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     2461 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     2521 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     2581 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     2641 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     2701 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     2761 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     2821 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     2881 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     2941 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     3001 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctgcaggc
     3061 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     3121 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     3181 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     3241 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     3301 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaacacgg
     3361 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     3421 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     3481 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     3541 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcatt
     3601 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     3661 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     3721 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     3781 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     3841 tagaccgaga tagggttgag tggccgctac agggcgctcc cattcgccat tcaggctgcg
     3901 caactgttgg gaagggcgtt tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     3961 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     4021 gtaaaacgac ggccagtgag cgcgacgtaa tacgactcac tatagggcga attggcggaa
     4081 ggccgtcaag gccgtgacgt taaaactatt aagccatcca atcgaccgtt agtcgaatca
     4141 ggaccgctgg tgcgagaagc cgcgaagtat ggcgaatgca tcgtataacg tgtggagtcc
     4201 gctcattaga gcgtcatgtt tagacaagaa agctacatat ttaattgatc ccgatgattt
     4261 tattgataaa ttgaccctaa ctccatacac ggtattctac aatggcgggg ttttggtcaa
     4321 aatttccgga ctgcgattgt acatgctgtt aacggctccg cccactatta atgaaattaa
     4381 aaattccaat tttaaaaaac gcagcaagag aaacatttgt atgaaagaat gcgtagaagg
     4441 aaagaaaaat gtcgtggaca tgctgaacaa caagattaat atgcctccgt gtataaaaaa
     4501 aatattgaac gatttgaaag aaaacaatgt accgcgcggc ggtatgtaca ggaagaggtt
     4561 tatactaaac tgttacattg caaacgtggt ttcgtgtgcc aagtgtgaaa accgatgttt
     4621 aatcaaggct ctgacgcatt tctacaacca cgactccaag tgtgtgggtg aagtcatgca
     4681 tcttttaatc aaatcccaag atgtgtataa accaccaaac tgccaaaaaa tgaaaactgt
     4741 ggacaagctc tgtccgtttg ctggcaactg caagggtctc aatcctattt gtaattattg
     4801 aataataaaa caattataaa tgctaaattt gttttttatt aacgatacaa accaaacgca
     4861 acaagaacat ttgtagtatt atctataatt gaaaacgcgt agttataatc gctgaggtaa
     4921 tatttaaaat cattttcaaa tgattcacag ttaatttgcg acaatataat tttattttca
     4981 cataaactag acgccttgtc gtcttcttct tcgtattcct tctctttttc atttttctcc
     5041 tcataaaaat taacatagtt attatcgtat ccatatatgt atctatcgta tagagtaaat
     5101 tttttgttgt cataaatata tatgtctttt ttaatggggt gtatagtacc gctgcgcata
     5161 gtttttctgt aatttacaac agtgctattt tctggtagtt cttcggagtg tgttgcttta
     5221 attattaaat ttatataatc aatgaatttg ggatcgtcgg ttttgtacaa tatgttgccg
     5281 gcatagtacg cagcttcttc tagttcaatt acaccatttt ttagcagcac cggattaaca
     5341 taactttcca aaatgttgta cgaaccgtta aacaaaaaca gttcacctcc cttttctata
     5401 ctattgtctg cgagcagttg tttgttgtta aaaataacag ccattgtaat gagacgcaca
     5461 aactaatatc aca
//
