LOCUS       sequence-224652-        5341 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for untagged recombinant protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5341)
  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 5341)
  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: pOCC82
FEATURES             Location/Qualifiers
     source          1..5341
                     /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"
     polyA_signal    807..941
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(844..863)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     898..917
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1014..1386
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(1738..1755)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1848..2436)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1928..1947)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2607..3398,3399..3467))
                     /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     3230..3249
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3469..3573)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3874..3896
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3888..3905
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3889..3905
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3916..3935
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3916..3934
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4545..4669
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(4707..5312)
                     /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 gtaatagaag cttgtcgaga agtactagag gatcataatc agccatacca
      721 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
      781 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
      841 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
      901 gtttgtccaa actcatcaat gtatcttatc atgtctggat ctgatcactg cttgagccta
      961 ggagatccga accagataag tgaaagcctg catccgcaga tcttaattaa taaaacacga
     1021 tacattgtta ttagtacatt tattaagcgc tagattctgt gcgttgttga tttacagaca
     1081 attgttgtac gtattttaat aattcattaa atttataatc tttagggtgg tatgttagag
     1141 cgaaaatcaa atgattttca gcgtctttat atctgaattt aaatattaaa tcctcaatag
     1201 atttgtaaaa taggtttcga ttagtttcaa acaagggttg tttttccgaa ccgatggctg
     1261 gactatctaa tggattttcg ctcaacgcca caaaacttgc caaatcttgt agcagcaatc
     1321 tagctttgtc gatattcgtt tgtgttttgt tttgtaataa aggttcgacg tcgttcaaaa
     1381 tattatgcgc ttttgtattt ctttcatcac tgtcgttagt gtacaattga ctcgacgtaa
     1441 acacgttaaa taacgcttgg acatatttaa catcgggcgt gttagcttta ttaggccgat
     1501 tatcgtcgtc gtcccaaccc tcgtcgttag aagttgcttc cgaagacgat tttgccatag
     1561 ccacacgacg cctattaatt gtgtcggcta acacgtccgc gatcaaattt gtagttgagc
     1621 tttttgggcc tcatgggcct tccgctcact gcccgctttc cagtcgggaa acctgtcgtg
     1681 ccagctgcat taacatggtc atagctgttt ccttgcgtat tgggcgctct ccgcttcctc
     1741 gctcactgac tcgctgcgct cggtcgttcg ggtaaagcct ggggtgccta atgagcaaaa
     1801 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     1861 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     1921 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     1981 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2041 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2101 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2161 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2221 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2281 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2341 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2401 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     2461 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     2521 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     2581 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     2641 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     2701 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     2761 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     2821 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     2881 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca
     2941 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     3001 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     3061 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     3121 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     3181 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg
     3241 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     3301 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     3361 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     3421 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcattac tcttcctttt
     3481 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     3541 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa
     3601 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     3661 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     3721 gggttgagtg gccgctacag ggcgctccca ttcgccattc aggctgcgca actgttggga
     3781 agggcgtttc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     3841 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     3901 ccagtgagcg cgacgtaata cgactcacta tagggcgaat tggcggaagg ccgtcaaggc
     3961 cgtgacgtta aaactattaa gccatccaat cgaccgttag tcgaatcagg accgctggtg
     4021 cgagaagccg cgaagtatgg cgaatgcatc gtataacgtg tggagtccgc tcattagagc
     4081 gtcatgttta gacaagaaag ctacatattt aattgatccc gatgatttta ttgataaatt
     4141 gaccctaact ccatacacgg tattctacaa tggcggggtt ttggtcaaaa tttccggact
     4201 gcgattgtac atgctgttaa cggctccgcc cactattaat gaaattaaaa attccaattt
     4261 taaaaaacgc agcaagagaa acatttgtat gaaagaatgc gtagaaggaa agaaaaatgt
     4321 cgtggacatg ctgaacaaca agattaatat gcctccgtgt ataaaaaaaa tattgaacga
     4381 tttgaaagaa aacaatgtac cgcgcggcgg tatgtacagg aagaggttta tactaaactg
     4441 ttacattgca aacgtggttt cgtgtgccaa gtgtgaaaac cgatgtttaa tcaaggctct
     4501 gacgcatttc tacaaccacg actccaagtg tgtgggtgaa gtcatgcatc ttttaatcaa
     4561 atcccaagat gtgtataaac caccaaactg ccaaaaaatg aaaactgtgg acaagctctg
     4621 tccgtttgct ggcaactgca agggtctcaa tcctatttgt aattattgaa taataaaaca
     4681 attataaatg ctaaatttgt tttttattaa cgatacaaac caaacgcaac aagaacattt
     4741 gtagtattat ctataattga aaacgcgtag ttataatcgc tgaggtaata tttaaaatca
     4801 ttttcaaatg attcacagtt aatttgcgac aatataattt tattttcaca taaactagac
     4861 gccttgtcgt cttcttcttc gtattccttc tctttttcat ttttctcctc ataaaaatta
     4921 acatagttat tatcgtatcc atatatgtat ctatcgtata gagtaaattt tttgttgtca
     4981 taaatatata tgtctttttt aatggggtgt atagtaccgc tgcgcatagt ttttctgtaa
     5041 tttacaacag tgctattttc tggtagttct tcggagtgtg ttgctttaat tattaaattt
     5101 atataatcaa tgaatttggg atcgtcggtt ttgtacaata tgttgccggc atagtacgca
     5161 gcttcttcta gttcaattac accatttttt agcagcaccg gattaacata actttccaaa
     5221 atgttgtacg aaccgttaaa caaaaacagt tcacctccct tttctatact attgtctgcg
     5281 agcagttgtt tgttgttaaa aataacagcc attgtaatga gacgcacaaa ctaatatcac
     5341 a
//
