LOCUS       sequence-224673-        6559 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with N-terminal HIS6-MBP tag, cleavable with
            3C protease.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6559)
  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 6559)
  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: pOCC102
FEATURES             Location/Qualifiers
     source          1..6559
                     /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             230..247
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             260..1360
                     /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     1334..1357
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1415..1438
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1547..1852
                     /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     1731..1750
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    2025..2159
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2062..2081)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2116..2135
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           2232..2604
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(2956..2973)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3066..3654)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3146..3165)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3825..4616,4617..4685))
                     /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     4448..4467
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4687..4791)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5092..5114
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5106..5123
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5107..5123
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5134..5153
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5134..5152
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5763..5887
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5925..6530)
                     /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 ggcagcagcc atcaccatca
      241 ccatcacagc agcggcagaa tgaaaatcga agaaggtaaa ctggtaatct ggattaacgg
      301 cgataaaggc tataacggtc tcgctgaagt cggtaagaaa ttcgagaaag ataccggaat
      361 taaagtcacc gttgagcatc cggataaact ggaagagaaa ttcccacagg ttgcggcaac
      421 tggcgatggc cctgacatta tcttctgggc acacgaccgc tttggtggct acgctcaatc
      481 tggcctgttg gctgaaatca ccccggacaa agcgttccag gacaagctgt atccgtttac
      541 ctgggatgcc gtacgttaca acggcaagct gattgcttac ccgatcgctg ttgaagcgtt
      601 atcgctgatt tataacaaag atctgctgcc gaacccgcca aaaacctggg aagagatccc
      661 ggcgctggat aaagaactga aagcgaaagg taagagcgcg ctgatgttca acctgcaaga
      721 accgtacttc acctggccgc tgattgctgc tgacgggggt tatgcgttca agtatgaaaa
      781 cggcaagtac gacattaaag acgtgggcgt ggataacgct ggcgcgaaag cgggtctgac
      841 cttcctggtt gacctgatta aaaacaaaca catgaatgca gacaccgatt actccatcgc
      901 agaagctgcc tttaataaag gcgaaacagc gatgaccatc aacggcccgt gggcatggtc
      961 caacatcgac accagcaaag tgaattatgg tgtaacggta ctgccgacct tcaagggtca
     1021 accatccaaa ccgttcgttg gcgtgctgag cgcaggtatt aacgccgcca gtccgaacaa
     1081 agagctggca aaagagttcc tcgaaaacta tctgctgact gatgaaggtc tggaagcggt
     1141 taataaagac aaaccgctgg gtgccgtagc gctgaagtct tacgaggaag agttggtgaa
     1201 agatccgcgt attgccgcca ctatggaaaa cgcccagaaa ggtgaaatca tgccgaacat
     1261 cccgcagatg tccgctttct ggtatgccgt gcgtactgcg gtgatcaacg ccgccagcgg
     1321 tcgtcagact gtcgatgaag ccctgaaaga cgcgcagact aattcgagct cgaacaacaa
     1381 caacaataac aataacaaca acagcagcgg cagactggaa gttctgttcc aggggccggc
     1441 ggccgcaagc gctatggcca caacctgtta cattgcacaa gataaaaata tatcatcacg
     1501 aacagtaaaa ctgtctgctt acataaacag taatacaagg ggtgttatgc agtttaaggt
     1561 ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga gtgatattat
     1621 tgacacgccc gggcgacgga tggtgatccc cctggccagt gcccgtctgc tgtcagataa
     1681 agtctcccgt gaactttacc cggtggtgca tatcggggat gaaagctggc gcatgatgac
     1741 caccgatatg gccagtgtgc cggtctccgt tatcggggaa gaagtggctg atctcagcca
     1801 ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat aaatgtcagg
     1861 ctcccttata cacagccagt ggcgcgccgt aatagaagct tgtcgagaag tactagagga
     1921 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     1981 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     2041 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     2101 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatct
     2161 gatcactgct tgagcctagg agatccgaac cagataagtg aaagcctgca tccgcagatc
     2221 ttaattaata aaacacgata cattgttatt agtacattta ttaagcgcta gattctgtgc
     2281 gttgttgatt tacagacaat tgttgtacgt attttaataa ttcattaaat ttataatctt
     2341 tagggtggta tgttagagcg aaaatcaaat gattttcagc gtctttatat ctgaatttaa
     2401 atattaaatc ctcaatagat ttgtaaaata ggtttcgatt agtttcaaac aagggttgtt
     2461 tttccgaacc gatggctgga ctatctaatg gattttcgct caacgccaca aaacttgcca
     2521 aatcttgtag cagcaatcta gctttgtcga tattcgtttg tgttttgttt tgtaataaag
     2581 gttcgacgtc gttcaaaata ttatgcgctt ttgtatttct ttcatcactg tcgttagtgt
     2641 acaattgact cgacgtaaac acgttaaata acgcttggac atatttaaca tcgggcgtgt
     2701 tagctttatt aggccgatta tcgtcgtcgt cccaaccctc gtcgttagaa gttgcttccg
     2761 aagacgattt tgccatagcc acacgacgcc tattaattgt gtcggctaac acgtccgcga
     2821 tcaaatttgt agttgagctt tttgggcctc atgggccttc cgctcactgc ccgctttcca
     2881 gtcgggaaac ctgtcgtgcc agctgcatta acatggtcat agctgtttcc ttgcgtattg
     2941 ggcgctctcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggg taaagcctgg
     3001 ggtgcctaat gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3061 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3121 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3181 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3241 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3301 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3361 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3421 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3481 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     3541 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3601 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3661 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3721 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3781 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3841 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3901 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3961 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     4021 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     4081 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     4141 gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     4201 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     4261 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     4321 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     4381 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     4441 acacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     4501 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     4561 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4621 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     4681 ctcattactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4741 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4801 ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     4861 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     4921 aaagaataga ccgagatagg gttgagtggc cgctacaggg cgctcccatt cgccattcag
     4981 gctgcgcaac tgttgggaag ggcgtttcgg tgcgggcctc ttcgctatta cgccagctgg
     5041 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     5101 gacgttgtaa aacgacggcc agtgagcgcg acgtaatacg actcactata gggcgaattg
     5161 gcggaaggcc gtcaaggccg tgacgttaaa actattaagc catccaatcg accgttagtc
     5221 gaatcaggac cgctggtgcg agaagccgcg aagtatggcg aatgcatcgt ataacgtgtg
     5281 gagtccgctc attagagcgt catgtttaga caagaaagct acatatttaa ttgatcccga
     5341 tgattttatt gataaattga ccctaactcc atacacggta ttctacaatg gcggggtttt
     5401 ggtcaaaatt tccggactgc gattgtacat gctgttaacg gctccgccca ctattaatga
     5461 aattaaaaat tccaatttta aaaaacgcag caagagaaac atttgtatga aagaatgcgt
     5521 agaaggaaag aaaaatgtcg tggacatgct gaacaacaag attaatatgc ctccgtgtat
     5581 aaaaaaaata ttgaacgatt tgaaagaaaa caatgtaccg cgcggcggta tgtacaggaa
     5641 gaggtttata ctaaactgtt acattgcaaa cgtggtttcg tgtgccaagt gtgaaaaccg
     5701 atgtttaatc aaggctctga cgcatttcta caaccacgac tccaagtgtg tgggtgaagt
     5761 catgcatctt ttaatcaaat cccaagatgt gtataaacca ccaaactgcc aaaaaatgaa
     5821 aactgtggac aagctctgtc cgtttgctgg caactgcaag ggtctcaatc ctatttgtaa
     5881 ttattgaata ataaaacaat tataaatgct aaatttgttt tttattaacg atacaaacca
     5941 aacgcaacaa gaacatttgt agtattatct ataattgaaa acgcgtagtt ataatcgctg
     6001 aggtaatatt taaaatcatt ttcaaatgat tcacagttaa tttgcgacaa tataatttta
     6061 ttttcacata aactagacgc cttgtcgtct tcttcttcgt attccttctc tttttcattt
     6121 ttctcctcat aaaaattaac atagttatta tcgtatccat atatgtatct atcgtataga
     6181 gtaaattttt tgttgtcata aatatatatg tcttttttaa tggggtgtat agtaccgctg
     6241 cgcatagttt ttctgtaatt tacaacagtg ctattttctg gtagttcttc ggagtgtgtt
     6301 gctttaatta ttaaatttat ataatcaatg aatttgggat cgtcggtttt gtacaatatg
     6361 ttgccggcat agtacgcagc ttcttctagt tcaattacac cattttttag cagcaccgga
     6421 ttaacataac tttccaaaat gttgtacgaa ccgttaaaca aaaacagttc acctcccttt
     6481 tctatactat tgtctgcgag cagttgtttg ttgttaaaaa taacagccat tgtaatgaga
     6541 cgcacaaact aatatcaca
//
