LOCUS       sequence-224672-        5983 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with C-terminal SNAP tag, cleavable with 
            TEV, and C-terminal HIS6 tag, cleavable with 3C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5983)
  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 5983)
  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: pOCC194
FEATURES             Location/Qualifiers
     source          1..5983
                     /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"
     regulatory      216..225
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             329..634
                     /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     513..532
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             690..710
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             711..1256
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     CDS             1257..1280
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1290..1307
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1449..1583
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1486..1505)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1540..1559
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1656..2028
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(2380..2397)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2490..3078)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2570..2589)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3249..4040,4041..4109))
                     /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     3872..3891
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4111..4215)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4516..4538
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4530..4547
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4531..4547
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4558..4577
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4558..4576
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5187..5311
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5349..5954)
                     /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 gcggccgcaa gcgctatggc
      241 cacaacctgt tacattgcac aagataaaaa tatatcatca cgaacagtaa aactgtctgc
      301 ttacataaac agtaatacaa ggggtgttat gcagtttaag gtttacacct ataaaagaga
      361 gagccgttat cgtctgtttg tggatgtaca gagtgatatt attgacacgc ccgggcgacg
      421 gatggtgatc cccctggcca gtgcccgtct gctgtcagat aaagtctccc gtgaacttta
      481 cccggtggtg catatcgggg atgaaagctg gcgcatgatg accaccgata tggccagtgt
      541 gccggtctcc gttatcgggg aagaagtggc tgatctcagc caccgcgaaa atgacatcaa
      601 aaacgccatt aacctgatgt tctggggaat ataaatgtca ggctccctta tacacagcca
      661 gtggcgcgcc gggatccagc agcggcagag aaaacttgta tttccagggc atggacaaag
      721 actgcgaaat gaagcgcacc accctggata gccctctggg caagctggaa ctgtctgggt
      781 gcgaacaggg cctgcaccgt atcatcttcc tgggcaaagg aacatctgcc gccgacgccg
      841 tggaagtgcc tgccccagcc gccgtgctgg gcggaccaga gccactgatg caggccaccg
      901 cctggctcaa cgcctacttt caccagcctg aggccatcga ggagttccct gtgccagccc
      961 tgcaccaccc agtgttccag caggagagct ttacccgcca ggtgctgtgg aaactgctga
     1021 aagtggtgaa gttcggagag gtcatcagct acagccacct ggccgccctg gccggcaatc
     1081 ccgccgccac cgccgccgtg aaaaccgccc tgagcggaaa tcccgtgccc attctgatcc
     1141 cctgccaccg ggtggtgcag ggcgacctgg acgtgggggg ctacgagggc gggctcgccg
     1201 tgaaagagtg gctgctggcc cacgagggcc acagactggg caagcctggg ctgggtctgg
     1261 aagttctgtt ccaggggccg ggcagcagcc atcaccatca ccatcacagt ggataataga
     1321 agcttgtcga gaagtactag aggatcataa tcagccatac cacatttgta gaggttttac
     1381 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     1441 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     1501 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     1561 atgtatctta tcatgtctgg atctgatcac tgcttgagcc taggagatcc gaaccagata
     1621 agtgaaagcc tgcatccgca gatcttaatt aataaaacac gatacattgt tattagtaca
     1681 tttattaagc gctagattct gtgcgttgtt gatttacaga caattgttgt acgtatttta
     1741 ataattcatt aaatttataa tctttagggt ggtatgttag agcgaaaatc aaatgatttt
     1801 cagcgtcttt atatctgaat ttaaatatta aatcctcaat agatttgtaa aataggtttc
     1861 gattagtttc aaacaagggt tgtttttccg aaccgatggc tggactatct aatggatttt
     1921 cgctcaacgc cacaaaactt gccaaatctt gtagcagcaa tctagctttg tcgatattcg
     1981 tttgtgtttt gttttgtaat aaaggttcga cgtcgttcaa aatattatgc gcttttgtat
     2041 ttctttcatc actgtcgtta gtgtacaatt gactcgacgt aaacacgtta aataacgctt
     2101 ggacatattt aacatcgggc gtgttagctt tattaggccg attatcgtcg tcgtcccaac
     2161 cctcgtcgtt agaagttgct tccgaagacg attttgccat agccacacga cgcctattaa
     2221 ttgtgtcggc taacacgtcc gcgatcaaat ttgtagttga gctttttggg cctcatgggc
     2281 cttccgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaacatgg
     2341 tcatagctgt ttccttgcgt attgggcgct ctccgcttcc tcgctcactg actcgctgcg
     2401 ctcggtcgtt cgggtaaagc ctggggtgcc taatgagcaa aaggccagca aaaggccagg
     2461 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     2521 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     2581 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2641 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     2701 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2761 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2821 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2881 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     2941 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3001 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3061 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3121 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3181 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3241 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3301 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     3361 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     3421 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     3481 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     3541 ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg
     3601 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     3661 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     3721 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     3781 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     3841 ccgagttgct cttgcccggc gtcaacacgg gataataccg cgccacatag cagaacttta
     3901 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     3961 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     4021 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     4081 agggcgacac ggaaatgttg aatactcatt actcttcctt tttcaatatt attgaagcat
     4141 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4201 aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt
     4261 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     4321 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tggccgctac
     4381 agggcgctcc cattcgccat tcaggctgcg caactgttgg gaagggcgtt tcggtgcggg
     4441 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     4501 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgacgtaa
     4561 tacgactcac tatagggcga attggcggaa ggccgtcaag gccgtgacgt taaaactatt
     4621 aagccatcca atcgaccgtt agtcgaatca ggaccgctgg tgcgagaagc cgcgaagtat
     4681 ggcgaatgca tcgtataacg tgtggagtcc gctcattaga gcgtcatgtt tagacaagaa
     4741 agctacatat ttaattgatc ccgatgattt tattgataaa ttgaccctaa ctccatacac
     4801 ggtattctac aatggcgggg ttttggtcaa aatttccgga ctgcgattgt acatgctgtt
     4861 aacggctccg cccactatta atgaaattaa aaattccaat tttaaaaaac gcagcaagag
     4921 aaacatttgt atgaaagaat gcgtagaagg aaagaaaaat gtcgtggaca tgctgaacaa
     4981 caagattaat atgcctccgt gtataaaaaa aatattgaac gatttgaaag aaaacaatgt
     5041 accgcgcggc ggtatgtaca ggaagaggtt tatactaaac tgttacattg caaacgtggt
     5101 ttcgtgtgcc aagtgtgaaa accgatgttt aatcaaggct ctgacgcatt tctacaacca
     5161 cgactccaag tgtgtgggtg aagtcatgca tcttttaatc aaatcccaag atgtgtataa
     5221 accaccaaac tgccaaaaaa tgaaaactgt ggacaagctc tgtccgtttg ctggcaactg
     5281 caagggtctc aatcctattt gtaattattg aataataaaa caattataaa tgctaaattt
     5341 gttttttatt aacgatacaa accaaacgca acaagaacat ttgtagtatt atctataatt
     5401 gaaaacgcgt agttataatc gctgaggtaa tatttaaaat cattttcaaa tgattcacag
     5461 ttaatttgcg acaatataat tttattttca cataaactag acgccttgtc gtcttcttct
     5521 tcgtattcct tctctttttc atttttctcc tcataaaaat taacatagtt attatcgtat
     5581 ccatatatgt atctatcgta tagagtaaat tttttgttgt cataaatata tatgtctttt
     5641 ttaatggggt gtatagtacc gctgcgcata gtttttctgt aatttacaac agtgctattt
     5701 tctggtagtt cttcggagtg tgttgcttta attattaaat ttatataatc aatgaatttg
     5761 ggatcgtcgg ttttgtacaa tatgttgccg gcatagtacg cagcttcttc tagttcaatt
     5821 acaccatttt ttagcagcac cggattaaca taactttcca aaatgttgta cgaaccgtta
     5881 aacaaaaaca gttcacctcc cttttctata ctattgtctg cgagcagttg tttgttgtta
     5941 aaaataacag ccattgtaat gagacgcaca aactaatatc aca
//
