LOCUS       sequence-224698-        5463 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with C-terminal HIS10, cleavable with 3C 
            protease and C-terminal YBBR tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5463)
  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 5463)
  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: pOCC219
FEATURES             Location/Qualifiers
     source          1..5463
                     /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             741..764
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             765..794
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     polyA_signal    929..1063
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(966..985)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1020..1039
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1136..1508
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(1860..1877)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1970..2558)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2050..2069)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2729..3520,3521..3589))
                     /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     3352..3371
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3591..3695)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3996..4018
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4010..4027
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4011..4027
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4038..4057
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4038..4056
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4667..4791
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(4829..5434)
                     /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 gggtagcgga tctggttcag acagtctgga gttcattgcg agcaagctcg
      721 ctggctcggg atccggttct ctcgaagttt tgttccaggg tccacatcat catcatcatc
      781 accaccatca ccactaaaga agcttgtcga gaagtactag aggatcataa tcagccatac
      841 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
      901 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
      961 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     1021 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atctgatcac tgcttgagcc
     1081 taggagatcc gaaccagata agtgaaagcc tgcatccgca gatcttaatt aataaaacac
     1141 gatacattgt tattagtaca tttattaagc gctagattct gtgcgttgtt gatttacaga
     1201 caattgttgt acgtatttta ataattcatt aaatttataa tctttagggt ggtatgttag
     1261 agcgaaaatc aaatgatttt cagcgtcttt atatctgaat ttaaatatta aatcctcaat
     1321 agatttgtaa aataggtttc gattagtttc aaacaagggt tgtttttccg aaccgatggc
     1381 tggactatct aatggatttt cgctcaacgc cacaaaactt gccaaatctt gtagcagcaa
     1441 tctagctttg tcgatattcg tttgtgtttt gttttgtaat aaaggttcga cgtcgttcaa
     1501 aatattatgc gcttttgtat ttctttcatc actgtcgtta gtgtacaatt gactcgacgt
     1561 aaacacgtta aataacgctt ggacatattt aacatcgggc gtgttagctt tattaggccg
     1621 attatcgtcg tcgtcccaac cctcgtcgtt agaagttgct tccgaagacg attttgccat
     1681 agccacacga cgcctattaa ttgtgtcggc taacacgtcc gcgatcaaat ttgtagttga
     1741 gctttttggg cctcatgggc cttccgctca ctgcccgctt tccagtcggg aaacctgtcg
     1801 tgccagctgc attaacatgg tcatagctgt ttccttgcgt attgggcgct ctccgcttcc
     1861 tcgctcactg actcgctgcg ctcggtcgtt cgggtaaagc ctggggtgcc taatgagcaa
     1921 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1981 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     2041 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     2101 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2161 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2221 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2281 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2341 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2401 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     2461 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     2521 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     2581 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     2641 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     2701 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     2761 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     2821 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     2881 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     2941 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     3001 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt
     3061 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     3121 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     3181 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     3241 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     3301 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaacacgg gataataccg
     3361 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     3421 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     3481 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     3541 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcatt actcttcctt
     3601 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     3661 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     3721 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     3781 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     3841 tagggttgag tggccgctac agggcgctcc cattcgccat tcaggctgcg caactgttgg
     3901 gaagggcgtt tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     3961 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     4021 ggccagtgag cgcgacgtaa tacgactcac tatagggcga attggcggaa ggccgtcaag
     4081 gccgtgacgt taaaactatt aagccatcca atcgaccgtt agtcgaatca ggaccgctgg
     4141 tgcgagaagc cgcgaagtat ggcgaatgca tcgtataacg tgtggagtcc gctcattaga
     4201 gcgtcatgtt tagacaagaa agctacatat ttaattgatc ccgatgattt tattgataaa
     4261 ttgaccctaa ctccatacac ggtattctac aatggcgggg ttttggtcaa aatttccgga
     4321 ctgcgattgt acatgctgtt aacggctccg cccactatta atgaaattaa aaattccaat
     4381 tttaaaaaac gcagcaagag aaacatttgt atgaaagaat gcgtagaagg aaagaaaaat
     4441 gtcgtggaca tgctgaacaa caagattaat atgcctccgt gtataaaaaa aatattgaac
     4501 gatttgaaag aaaacaatgt accgcgcggc ggtatgtaca ggaagaggtt tatactaaac
     4561 tgttacattg caaacgtggt ttcgtgtgcc aagtgtgaaa accgatgttt aatcaaggct
     4621 ctgacgcatt tctacaacca cgactccaag tgtgtgggtg aagtcatgca tcttttaatc
     4681 aaatcccaag atgtgtataa accaccaaac tgccaaaaaa tgaaaactgt ggacaagctc
     4741 tgtccgtttg ctggcaactg caagggtctc aatcctattt gtaattattg aataataaaa
     4801 caattataaa tgctaaattt gttttttatt aacgatacaa accaaacgca acaagaacat
     4861 ttgtagtatt atctataatt gaaaacgcgt agttataatc gctgaggtaa tatttaaaat
     4921 cattttcaaa tgattcacag ttaatttgcg acaatataat tttattttca cataaactag
     4981 acgccttgtc gtcttcttct tcgtattcct tctctttttc atttttctcc tcataaaaat
     5041 taacatagtt attatcgtat ccatatatgt atctatcgta tagagtaaat tttttgttgt
     5101 cataaatata tatgtctttt ttaatggggt gtatagtacc gctgcgcata gtttttctgt
     5161 aatttacaac agtgctattt tctggtagtt cttcggagtg tgttgcttta attattaaat
     5221 ttatataatc aatgaatttg ggatcgtcgg ttttgtacaa tatgttgccg gcatagtacg
     5281 cagcttcttc tagttcaatt acaccatttt ttagcagcac cggattaaca taactttcca
     5341 aaatgttgta cgaaccgtta aacaaaaaca gttcacctcc cttttctata ctattgtctg
     5401 cgagcagttg tttgttgtta aaaataacag ccattgtaat gagacgcaca aactaatatc
     5461 aca
//
