LOCUS       sequence-232308-        6138 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 6138)
  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 6138)
  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..6138
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..92
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     17..34
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     regulatory      134..143
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      138..147
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             251..556
                     /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     435..454
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             624..1334
                     /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             1335..1358
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1368..1385
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1527..1661
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1564..1583)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1618..1637
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1734..2106
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(2458..2475)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2568..3156)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2648..2667)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3327..4118,4119..4187))
                     /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     3950..3969
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4188..4292)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4593..4615
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4607..4624
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4608..4624
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4635..4654
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4635..4653
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5264..5388
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5426..6031)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
ORIGIN
        1 atcatggaga taattaaaat gataaccatc tcgcaaataa ataagtattt tactgttttc
       61 gtaacagttt tgtaataaaa aaacctataa atattccgga ttattcatac cgtcccacca
      121 tcgggcgcgg atcgccacca tggcggccgc aagcgctatg gccacaacct gttacattgc
      181 acaagataaa aatatatcat cacgaacagt aaaactgtct gcttacataa acagtaatac
      241 aaggggtgtt atgcagttta aggtttacac ctataaaaga gagagccgtt atcgtctgtt
      301 tgtggatgta cagagtgata ttattgacac gcccgggcga cggatggtga tccccctggc
      361 cagtgcccgt ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg
      421 ggatgaaagc tggcgcatga tgaccaccga tatggccagt gtgccggtct ccgttatcgg
      481 ggaagaagtg gctgatctca gccaccgcga aaatgacatc aaaaacgcca ttaacctgat
      541 gttctgggga atataaatgt caggctccct tatacacagc cagtggcgcg ccgggatccg
      601 ctggctccgc tgctggttct ggcatggtgt ctaagggcga agagctgatc aaggaaaata
      661 tgcgtatgaa ggtggtcatg gaaggttcgg tcaacggcca ccaattcaaa tgcacaggtg
      721 aaggagaagg cagaccgtac gagggaactc aaaccatgag gatcaaagtc atcgagggag
      781 gacccctgcc atttgccttt gacattcttg ccacgtcgtt catgtatggc agccgtactt
      841 ttatcaagta cccggccgac atccctgatt tctttaaaca gtcctttcct gagggtttta
      901 cttgggaaag agttacgaga tacgaagatg gtggagtcgt caccgtcacg caggacacca
      961 gccttgagga tggcgagctc gtctacaacg tcaaggtcag aggggtaaac tttccctcca
     1021 atggtcccgt gatgcagaag aagaccaagg gttgggagcc taatacagag atgatgtatc
     1081 cagcagatgg tggtctgaga ggatacactg acatcgcact gaaagttgat ggtggtggcc
     1141 atctgcactg caacttcgtg acaacttaca ggtcaaaaaa gaccgtcggg aacatcaaga
     1201 tgcccggtgt ccatgccgtt gatcaccgcc tggaaaggat cgaggagagt gacaatgaaa
     1261 cctacgtagt gcaacgcgaa gtggcagttg ccaaatacag caaccttggt ggtggcatgg
     1321 acgagctgta caagctggaa gttctgttcc aggggccggg cagcagccat caccatcacc
     1381 atcacagtgg ataatagaag cttgtcgaga agtactagag gatcataatc agccatacca
     1441 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     1501 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     1561 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     1621 gtttgtccaa actcatcaat gtatcttatc atgtctggat ctgatcactg cttgagccta
     1681 ggagatccga accagataag tgaaagcctg catccgcaga tcttaattaa taaaacacga
     1741 tacattgtta ttagtacatt tattaagcgc tagattctgt gcgttgttga tttacagaca
     1801 attgttgtac gtattttaat aattcattaa atttataatc tttagggtgg tatgttagag
     1861 cgaaaatcaa atgattttca gcgtctttat atctgaattt aaatattaaa tcctcaatag
     1921 atttgtaaaa taggtttcga ttagtttcaa acaagggttg tttttccgaa ccgatggctg
     1981 gactatctaa tggattttcg ctcaacgcca caaaacttgc caaatcttgt agcagcaatc
     2041 tagctttgtc gatattcgtt tgtgttttgt tttgtaataa aggttcgacg tcgttcaaaa
     2101 tattatgcgc ttttgtattt ctttcatcac tgtcgttagt gtacaattga ctcgacgtaa
     2161 acacgttaaa taacgcttgg acatatttaa catcgggcgt gttagcttta ttaggccgat
     2221 tatcgtcgtc gtcccaaccc tcgtcgttag aagttgcttc cgaagacgat tttgccatag
     2281 ccacacgacg cctattaatt gtgtcggcta acacgtccgc gatcaaattt gtagttgagc
     2341 tttttgggcc tcatgggcct tccgctcact gcccgctttc cagtcgggaa acctgtcgtg
     2401 ccagctgcat taacatggtc atagctgttt ccttgcgtat tgggcgctct ccgcttcctc
     2461 gctcactgac tcgctgcgct cggtcgttcg ggtaaagcct ggggtgccta atgagcaaaa
     2521 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2581 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2641 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2701 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2761 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2821 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2881 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2941 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3001 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3061 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3121 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3181 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3241 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3301 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3361 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3421 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga accacgctca
     3481 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     3541 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     3601 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     3661 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     3721 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     3781 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     3841 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     3901 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     3961 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     4021 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     4081 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     4141 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     4201 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     4261 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa
     4321 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     4381 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     4441 ggttgagtgg ccgctacagg gcgctcccat tcgccattca ggctgcgcaa ctgttgggaa
     4501 gggcgtttcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     4561 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     4621 cagtgagcgc gacgtaatac gactcactat agggcgaatt ggcggaaggc cgtcaaggcc
     4681 gtgacgttaa aactattaag ccatccaatc gaccgttagt cgaatcagga ccgctggtgc
     4741 gagaagccgc gaagtatggc gaatgcatcg tataacgtgt ggagtccgct cattagagcg
     4801 tcatgtttag acaagaaagc tacatattta attgatcccg atgattttat tgataaattg
     4861 accctaactc catacacggt attctacaat ggcggggttt tggtcaaaat ttccggactg
     4921 cgattgtaca tgctgttaac ggctccgccc actattaatg aaattaaaaa ttccaatttt
     4981 aaaaaacgca gcaagagaaa catttgtatg aaagaatgcg tagaaggaaa gaaaaatgtc
     5041 gtggacatgc tgaacaacaa gattaatatg cctccgtgta taaaaaaaat attgaacgat
     5101 ttgaaagaaa acaatgtacc gcgcggcggt atgtacagga agaggtttat actaaactgt
     5161 tacattgcaa acgtggtttc gtgtgccaag tgtgaaaacc gatgtttaat caaggctctg
     5221 acgcatttct acaaccacga ctccaagtgt gtgggtgaag tcatgcatct tttaatcaaa
     5281 tcccaagatg tgtataaacc accaaactgc caaaaaatga aaactgtgga caagctctgt
     5341 ccgtttgctg gcaactgcaa gggtctcaat cctatttgta attattgaat aataaaacaa
     5401 ttataaatgc taaatttgtt ttttattaac gatacaaacc aaacgcaaca agaacatttg
     5461 tagtattatc tataattgaa aacgcgtagt tataatcgct gaggtaatat ttaaaatcat
     5521 tttcaaatga ttcacagtta atttgcgaca atataatttt attttcacat aaactagacg
     5581 ccttgtcgtc ttcttcttcg tattccttct ctttttcatt tttctcctca taaaaattaa
     5641 catagttatt atcgtatcca tatatgtatc tatcgtatag agtaaatttt ttgttgtcat
     5701 aaatatatat gtctttttta atggggtgta tagtaccgct gcgcatagtt tttctgtaat
     5761 ttacaacagt gctattttct ggtagttctt cggagtgtgt tgctttaatt attaaattta
     5821 tataatcaat gaatttggga tcgtcggttt tgtacaatat gttgccggca tagtacgcag
     5881 cttcttctag ttcaattaca ccatttttta gcagcaccgg attaacataa ctttccaaaa
     5941 tgttgtacga accgttaaac aaaaacagtt cacctccctt ttctatacta ttgtctgcga
     6001 gcagttgttt gttgttaaaa ataacagcca ttgtaatgag acgcacaaac taatatcaca
     6061 agctgccacc attttgttcg cccaggactc tagctatagt tctagtggtt ggctacgtat
     6121 actccggaat attaatag
//
