LOCUS       sequence-232344-        6579 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 HIS6 tag, cleavable with 3C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6579)
  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 6579)
  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: pOCC28
FEATURES             Location/Qualifiers
     source          1..6579
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      42..51
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             54..1154
                     /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     1128..1151
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1209..1232
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1341..1646
                     /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     1525..1544
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             1684..1707
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1717..1734
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1876..2010
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1913..1932)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1967..1986
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           2083..2455
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(2807..2824)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2917..3505)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2997..3016)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3676..4467,4468..4536))
                     /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     4299..4318
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4537..4641)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4942..4964
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4956..4973
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4957..4973
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4984..5003
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4984..5002
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5613..5737
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5775..6380)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     promoter        6488..6579
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     6504..6521
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
ORIGIN
        1 attccggatt attcataccg tcccaccatc gggcgcggat cgccaccatg ggcatgaaaa
       61 tcgaagaagg taaactggta atctggatta acggcgataa aggctataac ggtctcgctg
      121 aagtcggtaa gaaattcgag aaagataccg gaattaaagt caccgttgag catccggata
      181 aactggaaga gaaattccca caggttgcgg caactggcga tggccctgac attatcttct
      241 gggcacacga ccgctttggt ggctacgctc aatctggcct gttggctgaa atcaccccgg
      301 acaaagcgtt ccaggacaag ctgtatccgt ttacctggga tgccgtacgt tacaacggca
      361 agctgattgc ttacccgatc gctgttgaag cgttatcgct gatttataac aaagatctgc
      421 tgccgaaccc gccaaaaacc tgggaagaga tcccggcgct ggataaagaa ctgaaagcga
      481 aaggtaagag cgcgctgatg ttcaacctgc aagaaccgta cttcacctgg ccgctgattg
      541 ctgctgacgg gggttatgcg ttcaagtatg aaaacggcaa gtacgacatt aaagacgtgg
      601 gcgtggataa cgctggcgcg aaagcgggtc tgaccttcct ggttgacctg attaaaaaca
      661 aacacatgaa tgcagacacc gattactcca tcgcagaagc tgcctttaat aaaggcgaaa
      721 cagcgatgac catcaacggc ccgtgggcat ggtccaacat cgacaccagc aaagtgaatt
      781 atggtgtaac ggtactgccg accttcaagg gtcaaccatc caaaccgttc gttggcgtgc
      841 tgagcgcagg tattaacgcc gccagtccga acaaagagct ggcaaaagag ttcctcgaaa
      901 actatctgct gactgatgaa ggtctggaag cggttaataa agacaaaccg ctgggtgccg
      961 tagcgctgaa gtcttacgag gaagagttgg tgaaagatcc gcgtattgcc gccactatgg
     1021 aaaacgccca gaaaggtgaa atcatgccga acatcccgca gatgtccgct ttctggtatg
     1081 ccgtgcgtac tgcggtgatc aacgccgcca gcggtcgtca gactgtcgat gaagccctga
     1141 aagacgcgca gactaattcg agctcgaaca acaacaacaa taacaataac aacaacagca
     1201 gcggcagact ggaagttctg ttccaggggc cggcggccgc aagcgctatg gccacaacct
     1261 gttacattgc acaagataaa aatatatcat cacgaacagt aaaactgtct gcttacataa
     1321 acagtaatac aaggggtgtt atgcagttta aggtttacac ctataaaaga gagagccgtt
     1381 atcgtctgtt tgtggatgta cagagtgata ttattgacac gcccgggcga cggatggtga
     1441 tccccctggc cagtgcccgt ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg
     1501 tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga tatggccagt gtgccggtct
     1561 ccgttatcgg ggaagaagtg gctgatctca gccaccgcga aaatgacatc aaaaacgcca
     1621 ttaacctgat gttctgggga atataaatgt caggctccct tatacacagc cagtggcgcg
     1681 ccgctggaag ttctgttcca ggggccgggc agcagccatc accatcacca tcacagtgga
     1741 taatagaagc ttgtcgagaa gtactagagg atcataatca gccataccac atttgtagag
     1801 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     1861 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     1921 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     1981 ctcatcaatg tatcttatca tgtctggatc tgatcactgc ttgagcctag gagatccgaa
     2041 ccagataagt gaaagcctgc atccgcagat cttaattaat aaaacacgat acattgttat
     2101 tagtacattt attaagcgct agattctgtg cgttgttgat ttacagacaa ttgttgtacg
     2161 tattttaata attcattaaa tttataatct ttagggtggt atgttagagc gaaaatcaaa
     2221 tgattttcag cgtctttata tctgaattta aatattaaat cctcaataga tttgtaaaat
     2281 aggtttcgat tagtttcaaa caagggttgt ttttccgaac cgatggctgg actatctaat
     2341 ggattttcgc tcaacgccac aaaacttgcc aaatcttgta gcagcaatct agctttgtcg
     2401 atattcgttt gtgttttgtt ttgtaataaa ggttcgacgt cgttcaaaat attatgcgct
     2461 tttgtatttc tttcatcact gtcgttagtg tacaattgac tcgacgtaaa cacgttaaat
     2521 aacgcttgga catatttaac atcgggcgtg ttagctttat taggccgatt atcgtcgtcg
     2581 tcccaaccct cgtcgttaga agttgcttcc gaagacgatt ttgccatagc cacacgacgc
     2641 ctattaattg tgtcggctaa cacgtccgcg atcaaatttg tagttgagct ttttgggcct
     2701 catgggcctt ccgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     2761 aacatggtca tagctgtttc cttgcgtatt gggcgctctc cgcttcctcg ctcactgact
     2821 cgctgcgctc ggtcgttcgg gtaaagcctg gggtgcctaa tgagcaaaag gccagcaaaa
     2881 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2941 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3001 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3061 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3121 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3181 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3241 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3301 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3361 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3421 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3481 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3541 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3601 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3661 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3721 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3781 cttaccatct ggccccagtg ctgcaatgat accgcgagaa ccacgctcac cggctccaga
     3841 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3901 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3961 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4021 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4081 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4141 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4201 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4261 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4321 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4381 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4441 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4501 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4561 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4621 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt
     4681 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     4741 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtggc
     4801 cgctacaggg cgctcccatt cgccattcag gctgcgcaac tgttgggaag ggcgtttcgg
     4861 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     4921 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg
     4981 acgtaatacg actcactata gggcgaattg gcggaaggcc gtcaaggccg tgacgttaaa
     5041 actattaagc catccaatcg accgttagtc gaatcaggac cgctggtgcg agaagccgcg
     5101 aagtatggcg aatgcatcgt ataacgtgtg gagtccgctc attagagcgt catgtttaga
     5161 caagaaagct acatatttaa ttgatcccga tgattttatt gataaattga ccctaactcc
     5221 atacacggta ttctacaatg gcggggtttt ggtcaaaatt tccggactgc gattgtacat
     5281 gctgttaacg gctccgccca ctattaatga aattaaaaat tccaatttta aaaaacgcag
     5341 caagagaaac atttgtatga aagaatgcgt agaaggaaag aaaaatgtcg tggacatgct
     5401 gaacaacaag attaatatgc ctccgtgtat aaaaaaaata ttgaacgatt tgaaagaaaa
     5461 caatgtaccg cgcggcggta tgtacaggaa gaggtttata ctaaactgtt acattgcaaa
     5521 cgtggtttcg tgtgccaagt gtgaaaaccg atgtttaatc aaggctctga cgcatttcta
     5581 caaccacgac tccaagtgtg tgggtgaagt catgcatctt ttaatcaaat cccaagatgt
     5641 gtataaacca ccaaactgcc aaaaaatgaa aactgtggac aagctctgtc cgtttgctgg
     5701 caactgcaag ggtctcaatc ctatttgtaa ttattgaata ataaaacaat tataaatgct
     5761 aaatttgttt tttattaacg atacaaacca aacgcaacaa gaacatttgt agtattatct
     5821 ataattgaaa acgcgtagtt ataatcgctg aggtaatatt taaaatcatt ttcaaatgat
     5881 tcacagttaa tttgcgacaa tataatttta ttttcacata aactagacgc cttgtcgtct
     5941 tcttcttcgt attccttctc tttttcattt ttctcctcat aaaaattaac atagttatta
     6001 tcgtatccat atatgtatct atcgtataga gtaaattttt tgttgtcata aatatatatg
     6061 tcttttttaa tggggtgtat agtaccgctg cgcatagttt ttctgtaatt tacaacagtg
     6121 ctattttctg gtagttcttc ggagtgtgtt gctttaatta ttaaatttat ataatcaatg
     6181 aatttgggat cgtcggtttt gtacaatatg ttgccggcat agtacgcagc ttcttctagt
     6241 tcaattacac cattttttag cagcaccgga ttaacataac tttccaaaat gttgtacgaa
     6301 ccgttaaaca aaaacagttc acctcccttt tctatactat tgtctgcgag cagttgtttg
     6361 ttgttaaaaa taacagccat tgtaatgaga cgcacaaact aatatcacaa gctgccacca
     6421 ttttgttcgc ccaggactct agctatagtt ctagtggttg gctacgtata ctccggaata
     6481 ttaatagatc atggagataa ttaaaatgat aaccatctcg caaataaata agtattttac
     6541 tgttttcgta acagttttgt aataaaaaaa cctataaat
//
