LOCUS       sequence-232315-        6558 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 6558)
  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 6558)
  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..6558
                     /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             60..77
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             90..1190
                     /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     1164..1187
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1245..1268
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1377..1682
                     /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     1561..1580
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    1855..1989
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1892..1911)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1946..1965
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           2062..2434
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(2786..2803)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2896..3484)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2976..2995)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3655..4446,4447..4515))
                     /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     4278..4297
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4516..4620)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4921..4943
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4935..4952
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4936..4952
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4963..4982
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4963..4981
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5592..5716
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5754..6359)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     promoter        6467..6558
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     6483..6500
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
ORIGIN
        1 attccggatt attcataccg tcccaccatc gggcgcggat cgccaccatg ggcagcagcc
       61 atcaccatca ccatcacagc agcggcagaa tgaaaatcga agaaggtaaa ctggtaatct
      121 ggattaacgg cgataaaggc tataacggtc tcgctgaagt cggtaagaaa ttcgagaaag
      181 ataccggaat taaagtcacc gttgagcatc cggataaact ggaagagaaa ttcccacagg
      241 ttgcggcaac tggcgatggc cctgacatta tcttctgggc acacgaccgc tttggtggct
      301 acgctcaatc tggcctgttg gctgaaatca ccccggacaa agcgttccag gacaagctgt
      361 atccgtttac ctgggatgcc gtacgttaca acggcaagct gattgcttac ccgatcgctg
      421 ttgaagcgtt atcgctgatt tataacaaag atctgctgcc gaacccgcca aaaacctggg
      481 aagagatccc ggcgctggat aaagaactga aagcgaaagg taagagcgcg ctgatgttca
      541 acctgcaaga accgtacttc acctggccgc tgattgctgc tgacgggggt tatgcgttca
      601 agtatgaaaa cggcaagtac gacattaaag acgtgggcgt ggataacgct ggcgcgaaag
      661 cgggtctgac cttcctggtt gacctgatta aaaacaaaca catgaatgca gacaccgatt
      721 actccatcgc agaagctgcc tttaataaag gcgaaacagc gatgaccatc aacggcccgt
      781 gggcatggtc caacatcgac accagcaaag tgaattatgg tgtaacggta ctgccgacct
      841 tcaagggtca accatccaaa ccgttcgttg gcgtgctgag cgcaggtatt aacgccgcca
      901 gtccgaacaa agagctggca aaagagttcc tcgaaaacta tctgctgact gatgaaggtc
      961 tggaagcggt taataaagac aaaccgctgg gtgccgtagc gctgaagtct tacgaggaag
     1021 agttggtgaa agatccgcgt attgccgcca ctatggaaaa cgcccagaaa ggtgaaatca
     1081 tgccgaacat cccgcagatg tccgctttct ggtatgccgt gcgtactgcg gtgatcaacg
     1141 ccgccagcgg tcgtcagact gtcgatgaag ccctgaaaga cgcgcagact aattcgagct
     1201 cgaacaacaa caacaataac aataacaaca acagcagcgg cagactggaa gttctgttcc
     1261 aggggccggc ggccgcaagc gctatggcca caacctgtta cattgcacaa gataaaaata
     1321 tatcatcacg aacagtaaaa ctgtctgctt acataaacag taatacaagg ggtgttatgc
     1381 agtttaaggt ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga
     1441 gtgatattat tgacacgccc gggcgacgga tggtgatccc cctggccagt gcccgtctgc
     1501 tgtcagataa agtctcccgt gaactttacc cggtggtgca tatcggggat gaaagctggc
     1561 gcatgatgac caccgatatg gccagtgtgc cggtctccgt tatcggggaa gaagtggctg
     1621 atctcagcca ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat
     1681 aaatgtcagg ctcccttata cacagccagt ggcgcgccgt aatagaagct tgtcgagaag
     1741 tactagagga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     1801 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     1861 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     1921 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     1981 gtctggatct gatcactgct tgagcctagg agatccgaac cagataagtg aaagcctgca
     2041 tccgcagatc ttaattaata aaacacgata cattgttatt agtacattta ttaagcgcta
     2101 gattctgtgc gttgttgatt tacagacaat tgttgtacgt attttaataa ttcattaaat
     2161 ttataatctt tagggtggta tgttagagcg aaaatcaaat gattttcagc gtctttatat
     2221 ctgaatttaa atattaaatc ctcaatagat ttgtaaaata ggtttcgatt agtttcaaac
     2281 aagggttgtt tttccgaacc gatggctgga ctatctaatg gattttcgct caacgccaca
     2341 aaacttgcca aatcttgtag cagcaatcta gctttgtcga tattcgtttg tgttttgttt
     2401 tgtaataaag gttcgacgtc gttcaaaata ttatgcgctt ttgtatttct ttcatcactg
     2461 tcgttagtgt acaattgact cgacgtaaac acgttaaata acgcttggac atatttaaca
     2521 tcgggcgtgt tagctttatt aggccgatta tcgtcgtcgt cccaaccctc gtcgttagaa
     2581 gttgcttccg aagacgattt tgccatagcc acacgacgcc tattaattgt gtcggctaac
     2641 acgtccgcga tcaaatttgt agttgagctt tttgggcctc atgggccttc cgctcactgc
     2701 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta acatggtcat agctgtttcc
     2761 ttgcgtattg ggcgctctcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggg
     2821 taaagcctgg ggtgcctaat gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2881 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2941 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3001 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3061 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3121 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3181 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3241 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3301 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     3361 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3421 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3481 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3541 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3601 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     3661 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     3721 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     3781 tgcaatgata ccgcgagaac cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     3841 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     3901 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     3961 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4021 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4081 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4141 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4201 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4261 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4321 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4381 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4441 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4501 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     4561 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     4621 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg
     4681 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     4741 tataaatcaa aagaatagac cgagataggg ttgagtggcc gctacagggc gctcccattc
     4801 gccattcagg ctgcgcaact gttgggaagg gcgtttcggt gcgggcctct tcgctattac
     4861 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     4921 cccagtcacg acgttgtaaa acgacggcca gtgagcgcga cgtaatacga ctcactatag
     4981 ggcgaattgg cggaaggccg tcaaggccgt gacgttaaaa ctattaagcc atccaatcga
     5041 ccgttagtcg aatcaggacc gctggtgcga gaagccgcga agtatggcga atgcatcgta
     5101 taacgtgtgg agtccgctca ttagagcgtc atgtttagac aagaaagcta catatttaat
     5161 tgatcccgat gattttattg ataaattgac cctaactcca tacacggtat tctacaatgg
     5221 cggggttttg gtcaaaattt ccggactgcg attgtacatg ctgttaacgg ctccgcccac
     5281 tattaatgaa attaaaaatt ccaattttaa aaaacgcagc aagagaaaca tttgtatgaa
     5341 agaatgcgta gaaggaaaga aaaatgtcgt ggacatgctg aacaacaaga ttaatatgcc
     5401 tccgtgtata aaaaaaatat tgaacgattt gaaagaaaac aatgtaccgc gcggcggtat
     5461 gtacaggaag aggtttatac taaactgtta cattgcaaac gtggtttcgt gtgccaagtg
     5521 tgaaaaccga tgtttaatca aggctctgac gcatttctac aaccacgact ccaagtgtgt
     5581 gggtgaagtc atgcatcttt taatcaaatc ccaagatgtg tataaaccac caaactgcca
     5641 aaaaatgaaa actgtggaca agctctgtcc gtttgctggc aactgcaagg gtctcaatcc
     5701 tatttgtaat tattgaataa taaaacaatt ataaatgcta aatttgtttt ttattaacga
     5761 tacaaaccaa acgcaacaag aacatttgta gtattatcta taattgaaaa cgcgtagtta
     5821 taatcgctga ggtaatattt aaaatcattt tcaaatgatt cacagttaat ttgcgacaat
     5881 ataattttat tttcacataa actagacgcc ttgtcgtctt cttcttcgta ttccttctct
     5941 ttttcatttt tctcctcata aaaattaaca tagttattat cgtatccata tatgtatcta
     6001 tcgtatagag taaatttttt gttgtcataa atatatatgt cttttttaat ggggtgtata
     6061 gtaccgctgc gcatagtttt tctgtaattt acaacagtgc tattttctgg tagttcttcg
     6121 gagtgtgttg ctttaattat taaatttata taatcaatga atttgggatc gtcggttttg
     6181 tacaatatgt tgccggcata gtacgcagct tcttctagtt caattacacc attttttagc
     6241 agcaccggat taacataact ttccaaaatg ttgtacgaac cgttaaacaa aaacagttca
     6301 cctccctttt ctatactatt gtctgcgagc agttgtttgt tgttaaaaat aacagccatt
     6361 gtaatgagac gcacaaacta atatcacaag ctgccaccat tttgttcgcc caggactcta
     6421 gctatagttc tagtggttgg ctacgtatac tccggaatat taatagatca tggagataat
     6481 taaaatgata accatctcgc aaataaataa gtattttact gttttcgtaa cagttttgta
     6541 ataaaaaaac ctataaat
//
