LOCUS       sequence-232249-        7479 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, and N-terminal HALO tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7479)
  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 7479)
  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: pOCC267
FEATURES             Location/Qualifiers
     source          1..7479
                     /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"
     CDS             152..169
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             182..1282
                     /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     1256..1279
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1337..1360
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1361..2251
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     CDS             2390..2695
                     /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     2574..2593
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    2868..3002
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2905..2924)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2959..2978
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           3075..3447
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(3799..3816)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3909..4497)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3989..4008)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4668..5459,5460..5528))
                     /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     5291..5310
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5529..5633)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5934..5956
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5948..5965
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5949..5965
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5976..5995
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5976..5994
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6605..6729
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(6767..7372)
                     /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 tgggcagcag ccatcaccat caccatcaca gcagcggcag
      181 aatgaaaatc gaagaaggta aactggtaat ctggattaac ggcgataaag gctataacgg
      241 tctcgctgaa gtcggtaaga aattcgagaa agataccgga attaaagtca ccgttgagca
      301 tccggataaa ctggaagaga aattcccaca ggttgcggca actggcgatg gccctgacat
      361 tatcttctgg gcacacgacc gctttggtgg ctacgctcaa tctggcctgt tggctgaaat
      421 caccccggac aaagcgttcc aggacaagct gtatccgttt acctgggatg ccgtacgtta
      481 caacggcaag ctgattgctt acccgatcgc tgttgaagcg ttatcgctga tttataacaa
      541 agatctgctg ccgaacccgc caaaaacctg ggaagagatc ccggcgctgg ataaagaact
      601 gaaagcgaaa ggtaagagcg cgctgatgtt caacctgcaa gaaccgtact tcacctggcc
      661 gctgattgct gctgacgggg gttatgcgtt caagtatgaa aacggcaagt acgacattaa
      721 agacgtgggc gtggataacg ctggcgcgaa agcgggtctg accttcctgg ttgacctgat
      781 taaaaacaaa cacatgaatg cagacaccga ttactccatc gcagaagctg cctttaataa
      841 aggcgaaaca gcgatgacca tcaacggccc gtgggcatgg tccaacatcg acaccagcaa
      901 agtgaattat ggtgtaacgg tactgccgac cttcaagggt caaccatcca aaccgttcgt
      961 tggcgtgctg agcgcaggta ttaacgccgc cagtccgaac aaagagctgg caaaagagtt
     1021 cctcgaaaac tatctgctga ctgatgaagg tctggaagcg gttaataaag acaaaccgct
     1081 gggtgccgta gcgctgaagt cttacgagga agagttggtg aaagatccgc gtattgccgc
     1141 cactatggaa aacgcccaga aaggtgaaat catgccgaac atcccgcaga tgtccgcttt
     1201 ctggtatgcc gtgcgtactg cggtgatcaa cgccgccagc ggtcgtcaga ctgtcgatga
     1261 agccctgaaa gacgcgcaga ctaattcgag ctcgaacaac aacaacaata acaataacaa
     1321 caacagcagc ggcagactgg aagttctgtt ccaggggccg atggctgaga tcggaaccgg
     1381 tttcccattc gacccgcatt acgtcgaggt tctgggagaa cgcatgcact acgtggatgt
     1441 cggaccacgt gacggtaccc cggtgttgtt cctgcatggt aaccctacta gttcgtacgt
     1501 ctggaggaac atcattcctc atgttgcccc cactcacaga tgtatcgcac ctgacctcat
     1561 tggcatggga aagagtgata aacccgactt gggttacttc ttcgatgacc acgtgcgttt
     1621 catggatgca ttcatcgagg cgctcggctt ggaggaagtc gttctggtca ttcatgactg
     1681 gggatctgca ctgggattcc actgggcaaa gaggaaccca gagagagtga aaggcatcgc
     1741 tttcatggaa ttcatcaggc caattccgac ttgggacgag tggcctgaat tcgcgagaga
     1801 aacattccag gctttccgca caacggatgt gggtcgtaag ctgatcattg accagaacgt
     1861 cttcatcgag ggtacactgc caatgggcgt ggtccgcccg ctgactgagg tggaaatgga
     1921 tcattaccgt gagccattcc tgaacccggt cgacagggaa cccctctgga gattccctaa
     1981 cgagttgccc atcgctggcg aaccagccaa cattgttgca ctcgtggagg aatacatgga
     2041 ctggttgcac caaagccctg ttcccaagtt gctgttctgg ggaaccccag gtgtgctgat
     2101 ccctcctgcc gaagcagcga ggttggcaaa gtctctgccc aactgtaaag ccgtggacat
     2161 cggtccaggc ctgaacctct tgcaggagga taaccctgac ctcatcggtt cagaaattgc
     2221 tagatggctg tcaactctcg agatcagtgg aggatccgct ggctccgctg ctggttctgg
     2281 cgcggccgca agcgctatgg ccacaacctg ttacattgca caagataaaa atatatcatc
     2341 acgaacagta aaactgtctg cttacataaa cagtaataca aggggtgtta tgcagtttaa
     2401 ggtttacacc tataaaagag agagccgtta tcgtctgttt gtggatgtac agagtgatat
     2461 tattgacacg cccgggcgac ggatggtgat ccccctggcc agtgcccgtc tgctgtcaga
     2521 taaagtctcc cgtgaacttt acccggtggt gcatatcggg gatgaaagct ggcgcatgat
     2581 gaccaccgat atggccagtg tgccggtctc cgttatcggg gaagaagtgg ctgatctcag
     2641 ccaccgcgaa aatgacatca aaaacgccat taacctgatg ttctggggaa tataaatgtc
     2701 aggctccctt atacacagcc agtggcgcgc cgtaatagaa gcttgtcgag aagtactaga
     2761 ggatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     2821 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     2881 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     2941 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     3001 tctgatcact gcttgagcct aggagatccg aaccagataa gtgaaagcct gcatccgcag
     3061 atcttaatta ataaaacacg atacattgtt attagtacat ttattaagcg ctagattctg
     3121 tgcgttgttg atttacagac aattgttgta cgtattttaa taattcatta aatttataat
     3181 ctttagggtg gtatgttaga gcgaaaatca aatgattttc agcgtcttta tatctgaatt
     3241 taaatattaa atcctcaata gatttgtaaa ataggtttcg attagtttca aacaagggtt
     3301 gtttttccga accgatggct ggactatcta atggattttc gctcaacgcc acaaaacttg
     3361 ccaaatcttg tagcagcaat ctagctttgt cgatattcgt ttgtgttttg ttttgtaata
     3421 aaggttcgac gtcgttcaaa atattatgcg cttttgtatt tctttcatca ctgtcgttag
     3481 tgtacaattg actcgacgta aacacgttaa ataacgcttg gacatattta acatcgggcg
     3541 tgttagcttt attaggccga ttatcgtcgt cgtcccaacc ctcgtcgtta gaagttgctt
     3601 ccgaagacga ttttgccata gccacacgac gcctattaat tgtgtcggct aacacgtccg
     3661 cgatcaaatt tgtagttgag ctttttgggc ctcatgggcc ttccgctcac tgcccgcttt
     3721 ccagtcggga aacctgtcgt gccagctgca ttaacatggt catagctgtt tccttgcgta
     3781 ttgggcgctc tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc gggtaaagcc
     3841 tggggtgcct aatgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3901 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3961 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4021 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4081 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4141 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4201 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4261 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4321 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     4381 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4441 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4501 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4561 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4621 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     4681 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     4741 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     4801 ataccgcgag aaccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4861 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     4921 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     4981 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     5041 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     5101 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     5161 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     5221 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     5281 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     5341 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     5401 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     5461 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     5521 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     5581 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     5641 ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     5701 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     5761 caaaagaata gaccgagata gggttgagtg gccgctacag ggcgctccca ttcgccattc
     5821 aggctgcgca actgttggga agggcgtttc ggtgcgggcc tcttcgctat tacgccagct
     5881 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     5941 acgacgttgt aaaacgacgg ccagtgagcg cgacgtaata cgactcacta tagggcgaat
     6001 tggcggaagg ccgtcaaggc cgtgacgtta aaactattaa gccatccaat cgaccgttag
     6061 tcgaatcagg accgctggtg cgagaagccg cgaagtatgg cgaatgcatc gtataacgtg
     6121 tggagtccgc tcattagagc gtcatgttta gacaagaaag ctacatattt aattgatccc
     6181 gatgatttta ttgataaatt gaccctaact ccatacacgg tattctacaa tggcggggtt
     6241 ttggtcaaaa tttccggact gcgattgtac atgctgttaa cggctccgcc cactattaat
     6301 gaaattaaaa attccaattt taaaaaacgc agcaagagaa acatttgtat gaaagaatgc
     6361 gtagaaggaa agaaaaatgt cgtggacatg ctgaacaaca agattaatat gcctccgtgt
     6421 ataaaaaaaa tattgaacga tttgaaagaa aacaatgtac cgcgcggcgg tatgtacagg
     6481 aagaggttta tactaaactg ttacattgca aacgtggttt cgtgtgccaa gtgtgaaaac
     6541 cgatgtttaa tcaaggctct gacgcatttc tacaaccacg actccaagtg tgtgggtgaa
     6601 gtcatgcatc ttttaatcaa atcccaagat gtgtataaac caccaaactg ccaaaaaatg
     6661 aaaactgtgg acaagctctg tccgtttgct ggcaactgca agggtctcaa tcctatttgt
     6721 aattattgaa taataaaaca attataaatg ctaaatttgt tttttattaa cgatacaaac
     6781 caaacgcaac aagaacattt gtagtattat ctataattga aaacgcgtag ttataatcgc
     6841 tgaggtaata tttaaaatca ttttcaaatg attcacagtt aatttgcgac aatataattt
     6901 tattttcaca taaactagac gccttgtcgt cttcttcttc gtattccttc tctttttcat
     6961 ttttctcctc ataaaaatta acatagttat tatcgtatcc atatatgtat ctatcgtata
     7021 gagtaaattt tttgttgtca taaatatata tgtctttttt aatggggtgt atagtaccgc
     7081 tgcgcatagt ttttctgtaa tttacaacag tgctattttc tggtagttct tcggagtgtg
     7141 ttgctttaat tattaaattt atataatcaa tgaatttggg atcgtcggtt ttgtacaata
     7201 tgttgccggc atagtacgca gcttcttcta gttcaattac accatttttt agcagcaccg
     7261 gattaacata actttccaaa atgttgtacg aaccgttaaa caaaaacagt tcacctccct
     7321 tttctatact attgtctgcg agcagttgtt tgttgttaaa aataacagcc attgtaatga
     7381 gacgcacaaa ctaatatcac aagctgccac cattttgttc gcccaggact ctagctatag
     7441 ttctagtggt tggctacgta tactccggaa tattaatag
//
