LOCUS       sequence-224684-        7306 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with N-terminal MBP tag, cleavable with 3C 
            and C-terminal mKate2, and HIS6, cleavable with 3C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7306)
  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 7306)
  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: pOCC204
FEATURES             Location/Qualifiers
     source          1..7306
                     /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"
     CDS             224..1324
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELVKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     1298..1321
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1379..1402
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1511..1816
                     /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     1695..1714
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             1884..2579
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     CDS             2580..2603
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2613..2630
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2772..2906
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2809..2828)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2863..2882
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           2979..3351
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(3703..3720)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3813..4401)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3893..3912)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4572..5363,5364..5432))
                     /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     5195..5214
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5434..5538)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5839..5861
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5853..5870
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5854..5870
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5881..5900
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5881..5899
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6510..6634
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(6672..7277)
                     /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 ggcatgaaaa tcgaagaagg
      241 taaactggta atctggatta acggcgataa aggctataac ggtctcgctg aagtcggtaa
      301 gaaattcgag aaagataccg gaattaaagt caccgttgag catccggata aactggaaga
      361 gaaattccca caggttgcgg caactggcga tggccctgac attatcttct gggcacacga
      421 ccgctttggt ggctacgctc aatctggcct gttggctgaa atcaccccgg acaaagcgtt
      481 ccaggacaag ctgtatccgt ttacctggga tgccgtacgt tacaacggca agctgattgc
      541 ttacccgatc gctgttgaag cgttatcgct gatttataac aaagatctgc tgccgaaccc
      601 gccaaaaacc tgggaagaga tcccggcgct ggataaagaa ctgaaagcga aaggtaagag
      661 cgcgctgatg ttcaacctgc aagaaccgta cttcacctgg ccgctgattg ctgctgacgg
      721 gggttatgcg ttcaagtatg aaaacggcaa gtacgacatt aaagacgtgg gcgtggataa
      781 cgctggcgcg aaagcgggtc tgaccttcct ggttgacctg attaaaaaca aacacatgaa
      841 tgcagacacc gattactcca tcgcagaagc tgcctttaat aaaggcgaaa cagcgatgac
      901 catcaacggc ccgtgggcat ggtccaacat cgacaccagc aaagtgaatt atggtgtaac
      961 ggtactgccg accttcaagg gtcaaccatc caaaccgttc gttggcgtgc tgagcgcagg
     1021 tattaacgcc gccagtccga acaaagagct ggcaaaagag ttcctcgaaa actatctgct
     1081 gactgatgaa ggtctggaag cggttaataa agacaaaccg ctgggtgccg tagcgctgaa
     1141 gtcttacgag gaagagttgg tgaaagatcc gcgtattgcc gccactatgg aaaacgccca
     1201 gaaaggtgaa atcatgccga acatcccgca gatgtccgct ttctggtatg ccgtgcgtac
     1261 tgcggtgatc aacgccgcca gcggtcgtca gactgtcgat gaagccctga aagacgcgca
     1321 gactaattcg agctcgaaca acaacaacaa taacaataac aacaacagca gcggcagact
     1381 ggaagttctg ttccaggggc cggcggccgc aagcgctatg gccacaacct gttacattgc
     1441 acaagataaa aatatatcat cacgaacagt aaaactgtct gcttacataa acagtaatac
     1501 aaggggtgtt atgcagttta aggtttacac ctataaaaga gagagccgtt atcgtctgtt
     1561 tgtggatgta cagagtgata ttattgacac gcccgggcga cggatggtga tccccctggc
     1621 cagtgcccgt ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg
     1681 ggatgaaagc tggcgcatga tgaccaccga tatggccagt gtgccggtct ccgttatcgg
     1741 ggaagaagtg gctgatctca gccaccgcga aaatgacatc aaaaacgcca ttaacctgat
     1801 gttctgggga atataaatgt caggctccct tatacacagc cagtggcgcg ccgggatccg
     1861 ctggctccgc tgctggttct ggcatggtga gcgagctgat taaggagaac atgcacatga
     1921 agctgtacat ggagggcacc gtgaacaacc accacttcaa gtgcacatcc gagggcgaag
     1981 gcaagcccta cgagggcacc cagaccatga gaatcaaggc ggtcgagggc ggccctctcc
     2041 ccttcgcctt cgacatcctg gctaccagct tcatgtacgg cagcaaaacc ttcatcaacc
     2101 acacccaggg catccccgac ttctttaagc agtccttccc cgagggcttc acatgggaga
     2161 gagtcaccac atacgaagac gggggcgtgc tgaccgctac ccaggacacc agcctccagg
     2221 acggctgcct catctacaac gtcaagatca gaggggtgaa cttcccatcc aacggccctg
     2281 tgatgcagaa gaaaacactc ggctgggagg cctccaccga gaccctgtac cccgctgacg
     2341 gcggcctgga aggcagagcc gacatggccc tgaagctcgt gggcgggggc cacctgatct
     2401 gcaacttgaa gaccacatac agatccaaga aacccgctaa gaacctcaag atgcccggcg
     2461 tctactatgt ggacagaaga ctggaaagaa tcaaggaggc cgacaaagag acctacgtcg
     2521 agcagcacga ggtggctgtg gccagatact gcgacctccc tagcaaactg gggcacagac
     2581 tggaagttct gttccagggg ccgggcagca gccatcacca tcaccatcac agtggataat
     2641 agaagcttgt cgagaagtac tagaggatca taatcagcca taccacattt gtagaggttt
     2701 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     2761 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     2821 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     2881 tcaatgtatc ttatcatgtc tggatctgat cactgcttga gcctaggaga tccgaaccag
     2941 ataagtgaaa gcctgcatcc gcagatctta attaataaaa cacgatacat tgttattagt
     3001 acatttatta agcgctagat tctgtgcgtt gttgatttac agacaattgt tgtacgtatt
     3061 ttaataattc attaaattta taatctttag ggtggtatgt tagagcgaaa atcaaatgat
     3121 tttcagcgtc tttatatctg aatttaaata ttaaatcctc aatagatttg taaaataggt
     3181 ttcgattagt ttcaaacaag ggttgttttt ccgaaccgat ggctggacta tctaatggat
     3241 tttcgctcaa cgccacaaaa cttgccaaat cttgtagcag caatctagct ttgtcgatat
     3301 tcgtttgtgt tttgttttgt aataaaggtt cgacgtcgtt caaaatatta tgcgcttttg
     3361 tatttctttc atcactgtcg ttagtgtaca attgactcga cgtaaacacg ttaaataacg
     3421 cttggacata tttaacatcg ggcgtgttag ctttattagg ccgattatcg tcgtcgtccc
     3481 aaccctcgtc gttagaagtt gcttccgaag acgattttgc catagccaca cgacgcctat
     3541 taattgtgtc ggctaacacg tccgcgatca aatttgtagt tgagcttttt gggcctcatg
     3601 ggccttccgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaaca
     3661 tggtcatagc tgtttccttg cgtattgggc gctctccgct tcctcgctca ctgactcgct
     3721 gcgctcggtc gttcgggtaa agcctggggt gcctaatgag caaaaggcca gcaaaaggcc
     3781 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3841 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3901 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3961 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4021 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4081 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4141 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4201 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4261 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4321 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4381 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4441 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4501 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4561 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4621 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4681 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     4741 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     4801 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4861 agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt
     4921 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     4981 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5041 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5101 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5161 cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact
     5221 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5281 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5341 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5401 ataagggcga cacggaaatg ttgaatactc attactcttc ctttttcaat attattgaag
     5461 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5521 acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat
     5581 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     5641 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtggccgc
     5701 tacagggcgc tcccattcgc cattcaggct gcgcaactgt tgggaagggc gtttcggtgc
     5761 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     5821 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gagcgcgacg
     5881 taatacgact cactataggg cgaattggcg gaaggccgtc aaggccgtga cgttaaaact
     5941 attaagccat ccaatcgacc gttagtcgaa tcaggaccgc tggtgcgaga agccgcgaag
     6001 tatggcgaat gcatcgtata acgtgtggag tccgctcatt agagcgtcat gtttagacaa
     6061 gaaagctaca tatttaattg atcccgatga ttttattgat aaattgaccc taactccata
     6121 cacggtattc tacaatggcg gggttttggt caaaatttcc ggactgcgat tgtacatgct
     6181 gttaacggct ccgcccacta ttaatgaaat taaaaattcc aattttaaaa aacgcagcaa
     6241 gagaaacatt tgtatgaaag aatgcgtaga aggaaagaaa aatgtcgtgg acatgctgaa
     6301 caacaagatt aatatgcctc cgtgtataaa aaaaatattg aacgatttga aagaaaacaa
     6361 tgtaccgcgc ggcggtatgt acaggaagag gtttatacta aactgttaca ttgcaaacgt
     6421 ggtttcgtgt gccaagtgtg aaaaccgatg tttaatcaag gctctgacgc atttctacaa
     6481 ccacgactcc aagtgtgtgg gtgaagtcat gcatctttta atcaaatccc aagatgtgta
     6541 taaaccacca aactgccaaa aaatgaaaac tgtggacaag ctctgtccgt ttgctggcaa
     6601 ctgcaagggt ctcaatccta tttgtaatta ttgaataata aaacaattat aaatgctaaa
     6661 tttgtttttt attaacgata caaaccaaac gcaacaagaa catttgtagt attatctata
     6721 attgaaaacg cgtagttata atcgctgagg taatatttaa aatcattttc aaatgattca
     6781 cagttaattt gcgacaatat aattttattt tcacataaac tagacgcctt gtcgtcttct
     6841 tcttcgtatt ccttctcttt ttcatttttc tcctcataaa aattaacata gttattatcg
     6901 tatccatata tgtatctatc gtatagagta aattttttgt tgtcataaat atatatgtct
     6961 tttttaatgg ggtgtatagt accgctgcgc atagtttttc tgtaatttac aacagtgcta
     7021 ttttctggta gttcttcgga gtgtgttgct ttaattatta aatttatata atcaatgaat
     7081 ttgggatcgt cggttttgta caatatgttg ccggcatagt acgcagcttc ttctagttca
     7141 attacaccat tttttagcag caccggatta acataacttt ccaaaatgtt gtacgaaccg
     7201 ttaaacaaaa acagttcacc tcccttttct atactattgt ctgcgagcag ttgtttgttg
     7261 ttaaaaataa cagccattgt aatgagacgc acaaactaat atcaca
//
