LOCUS       sequence-224671-        6139 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with C-terminal mRuby3, and C-terminal HIS6 
            tag, cleavable with 3C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6139)
  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 6139)
  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: pOCC292
FEATURES             Location/Qualifiers
     source          1..6139
                     /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             702..1412
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
                     /label=mRuby3
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTD
                     IALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAV
                     AKYSNLGGGMDELYK"
     CDS             1413..1436
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1446..1463
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1605..1739
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1642..1661)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1696..1715
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1812..2184
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(2536..2553)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2646..3234)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2726..2745)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3405..4196,4197..4265))
                     /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     4028..4047
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4267..4371)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4672..4694
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4686..4703
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4687..4703
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4714..4733
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4714..4732
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5343..5467
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5505..6110)
                     /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 gggatccgct ggctccgctg ctggttctgg catggtgtct aagggcgaag
      721 agctgatcaa ggaaaatatg cgtatgaagg tggtcatgga aggttcggtc aacggccacc
      781 aattcaaatg cacaggtgaa ggagaaggca gaccgtacga gggaactcaa accatgagga
      841 tcaaagtcat cgagggagga cccctgccat ttgcctttga cattcttgcc acgtcgttca
      901 tgtatggcag ccgtactttt atcaagtacc cggccgacat ccctgatttc tttaaacagt
      961 cctttcctga gggttttact tgggaaagag ttacgagata cgaagatggt ggagtcgtca
     1021 ccgtcacgca ggacaccagc cttgaggatg gcgagctcgt ctacaacgtc aaggtcagag
     1081 gggtaaactt tccctccaat ggtcccgtga tgcagaagaa gaccaagggt tgggagccta
     1141 atacagagat gatgtatcca gcagatggtg gtctgagagg atacactgac atcgcactga
     1201 aagttgatgg tggtggccat ctgcactgca acttcgtgac aacttacagg tcaaaaaaga
     1261 ccgtcgggaa catcaagatg cccggtgtcc atgccgttga tcaccgcctg gaaaggatcg
     1321 aggagagtga caatgaaacc tacgtagtgc aacgcgaagt ggcagttgcc aaatacagca
     1381 accttggtgg tggcatggac gagctgtaca agctggaagt tctgttccag gggccgggca
     1441 gcagccatca ccatcaccat cacagtggat aatagaagct tgtcgagaag tactagagga
     1501 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     1561 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     1621 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     1681 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatct
     1741 gatcactgct tgagcctagg agatccgaac cagataagtg aaagcctgca tccgcagatc
     1801 ttaattaata aaacacgata cattgttatt agtacattta ttaagcgcta gattctgtgc
     1861 gttgttgatt tacagacaat tgttgtacgt attttaataa ttcattaaat ttataatctt
     1921 tagggtggta tgttagagcg aaaatcaaat gattttcagc gtctttatat ctgaatttaa
     1981 atattaaatc ctcaatagat ttgtaaaata ggtttcgatt agtttcaaac aagggttgtt
     2041 tttccgaacc gatggctgga ctatctaatg gattttcgct caacgccaca aaacttgcca
     2101 aatcttgtag cagcaatcta gctttgtcga tattcgtttg tgttttgttt tgtaataaag
     2161 gttcgacgtc gttcaaaata ttatgcgctt ttgtatttct ttcatcactg tcgttagtgt
     2221 acaattgact cgacgtaaac acgttaaata acgcttggac atatttaaca tcgggcgtgt
     2281 tagctttatt aggccgatta tcgtcgtcgt cccaaccctc gtcgttagaa gttgcttccg
     2341 aagacgattt tgccatagcc acacgacgcc tattaattgt gtcggctaac acgtccgcga
     2401 tcaaatttgt agttgagctt tttgggcctc atgggccttc cgctcactgc ccgctttcca
     2461 gtcgggaaac ctgtcgtgcc agctgcatta acatggtcat agctgtttcc ttgcgtattg
     2521 ggcgctctcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggg taaagcctgg
     2581 ggtgcctaat gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2641 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2701 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2761 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2821 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2881 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2941 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3001 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3061 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     3121 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3181 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3241 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3301 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3361 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3421 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3481 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3541 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3601 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3661 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3721 gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3781 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     3841 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     3901 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     3961 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     4021 acacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     4081 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     4141 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4201 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     4261 ctcattactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4321 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4381 ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     4441 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     4501 aaagaataga ccgagatagg gttgagtggc cgctacaggg cgctcccatt cgccattcag
     4561 gctgcgcaac tgttgggaag ggcgtttcgg tgcgggcctc ttcgctatta cgccagctgg
     4621 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     4681 gacgttgtaa aacgacggcc agtgagcgcg acgtaatacg actcactata gggcgaattg
     4741 gcggaaggcc gtcaaggccg tgacgttaaa actattaagc catccaatcg accgttagtc
     4801 gaatcaggac cgctggtgcg agaagccgcg aagtatggcg aatgcatcgt ataacgtgtg
     4861 gagtccgctc attagagcgt catgtttaga caagaaagct acatatttaa ttgatcccga
     4921 tgattttatt gataaattga ccctaactcc atacacggta ttctacaatg gcggggtttt
     4981 ggtcaaaatt tccggactgc gattgtacat gctgttaacg gctccgccca ctattaatga
     5041 aattaaaaat tccaatttta aaaaacgcag caagagaaac atttgtatga aagaatgcgt
     5101 agaaggaaag aaaaatgtcg tggacatgct gaacaacaag attaatatgc ctccgtgtat
     5161 aaaaaaaata ttgaacgatt tgaaagaaaa caatgtaccg cgcggcggta tgtacaggaa
     5221 gaggtttata ctaaactgtt acattgcaaa cgtggtttcg tgtgccaagt gtgaaaaccg
     5281 atgtttaatc aaggctctga cgcatttcta caaccacgac tccaagtgtg tgggtgaagt
     5341 catgcatctt ttaatcaaat cccaagatgt gtataaacca ccaaactgcc aaaaaatgaa
     5401 aactgtggac aagctctgtc cgtttgctgg caactgcaag ggtctcaatc ctatttgtaa
     5461 ttattgaata ataaaacaat tataaatgct aaatttgttt tttattaacg atacaaacca
     5521 aacgcaacaa gaacatttgt agtattatct ataattgaaa acgcgtagtt ataatcgctg
     5581 aggtaatatt taaaatcatt ttcaaatgat tcacagttaa tttgcgacaa tataatttta
     5641 ttttcacata aactagacgc cttgtcgtct tcttcttcgt attccttctc tttttcattt
     5701 ttctcctcat aaaaattaac atagttatta tcgtatccat atatgtatct atcgtataga
     5761 gtaaattttt tgttgtcata aatatatatg tcttttttaa tggggtgtat agtaccgctg
     5821 cgcatagttt ttctgtaatt tacaacagtg ctattttctg gtagttcttc ggagtgtgtt
     5881 gctttaatta ttaaatttat ataatcaatg aatttgggat cgtcggtttt gtacaatatg
     5941 ttgccggcat agtacgcagc ttcttctagt tcaattacac cattttttag cagcaccgga
     6001 ttaacataac tttccaaaat gttgtacgaa ccgttaaaca aaaacagttc acctcccttt
     6061 tctatactat tgtctgcgag cagttgtttg ttgttaaaaa taacagccat tgtaatgaga
     6121 cgcacaaact aatatcaca
//
