LOCUS       sequence-232352-        6126 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with N-terminal HIS6 tag, cleavable with 3C,
            and C-terminal tagRFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6126)
  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 6126)
  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: pOCC17
FEATURES             Location/Qualifiers
     source          1..6126
                     /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..113
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             222..527
                     /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     406..425
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             595..1290
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSDMALK
                     LVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAVARYC
                     DLPSKLGHK"
     polyA_signal    1423..1557
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1460..1479)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1514..1533
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1630..2002
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(2354..2371)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2464..3052)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2544..2563)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3223..4014,4015..4083))
                     /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     3846..3865
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4084..4188)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4489..4511
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4503..4520
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4504..4520
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4531..4550
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4531..4549
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5160..5284
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5322..5927)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     promoter        6035..6126
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     6051..6068
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
ORIGIN
        1 attccggatt attcataccg tcccaccatc gggcgcggat cgccaccatg ggcagcagcc
       61 atcaccatca ccatcacagc agcggcagac tggaagttct gttccagggg ccggcggccg
      121 caagcgctat ggccacaacc tgttacattg cacaagataa aaatatatca tcacgaacag
      181 taaaactgtc tgcttacata aacagtaata caaggggtgt tatgcagttt aaggtttaca
      241 cctataaaag agagagccgt tatcgtctgt ttgtggatgt acagagtgat attattgaca
      301 cgcccgggcg acggatggtg atccccctgg ccagtgcccg tctgctgtca gataaagtct
      361 cccgtgaact ttacccggtg gtgcatatcg gggatgaaag ctggcgcatg atgaccaccg
      421 atatggccag tgtgccggtc tccgttatcg gggaagaagt ggctgatctc agccaccgcg
      481 aaaatgacat caaaaacgcc attaacctga tgttctgggg aatataaatg tcaggctccc
      541 ttatacacag ccagtggcgc gccgggatcc gctggctccg ctgctggttc tggcatgagc
      601 gagctgatta aggagaacat gcacatgaag ctgtacatgg agggcaccgt gaacaaccac
      661 cacttcaagt gcacatccga gggcgaaggc aagccctacg agggcaccca gaccatgaga
      721 atcaaggtgg tcgagggcgg ccctctcccc ttcgccttcg acatcctggc taccagcttc
      781 atgtacggca gcagaacctt catcaaccac acccagggca tccccgactt ctttaagcag
      841 tccttccctg agggcttcac atgggagaga gtcaccacat acgaagacgg gggcgtgctg
      901 accgctaccc aggacaccag cctccaggac ggctgcctca tctacaacgt caagatcaga
      961 ggggtgaact tcccatccaa cggccctgtg atgcagaaga aaacactcgg ctgggaggcc
     1021 aacaccgaga tgctgtaccc cgctgacggc ggcctggaag gcagaagcga catggccctg
     1081 aagctcgtgg gcgggggcca cctgatctgc aacttcaaga ccacatacag atccaagaaa
     1141 cccgctaaga acctcaagat gcccggcgtc tactatgtgg accacagact ggaaagaatc
     1201 aaggaggccg acaaagagac ctacgtcgag cagcacgagg tggctgtggc cagatactgc
     1261 gacctcccta gcaaactggg gcacaagtaa tagaagcttg tcgagaagta ctagaggatc
     1321 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     1381 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     1441 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     1501 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatctga
     1561 tcactgcttg agcctaggag atccgaacca gataagtgaa agcctgcatc cgcagatctt
     1621 aattaataaa acacgataca ttgttattag tacatttatt aagcgctaga ttctgtgcgt
     1681 tgttgattta cagacaattg ttgtacgtat tttaataatt cattaaattt ataatcttta
     1741 gggtggtatg ttagagcgaa aatcaaatga ttttcagcgt ctttatatct gaatttaaat
     1801 attaaatcct caatagattt gtaaaatagg tttcgattag tttcaaacaa gggttgtttt
     1861 tccgaaccga tggctggact atctaatgga ttttcgctca acgccacaaa acttgccaaa
     1921 tcttgtagca gcaatctagc tttgtcgata ttcgtttgtg ttttgttttg taataaaggt
     1981 tcgacgtcgt tcaaaatatt atgcgctttt gtatttcttt catcactgtc gttagtgtac
     2041 aattgactcg acgtaaacac gttaaataac gcttggacat atttaacatc gggcgtgtta
     2101 gctttattag gccgattatc gtcgtcgtcc caaccctcgt cgttagaagt tgcttccgaa
     2161 gacgattttg ccatagccac acgacgccta ttaattgtgt cggctaacac gtccgcgatc
     2221 aaatttgtag ttgagctttt tgggcctcat gggccttccg ctcactgccc gctttccagt
     2281 cgggaaacct gtcgtgccag ctgcattaac atggtcatag ctgtttcctt gcgtattggg
     2341 cgctctccgc ttcctcgctc actgactcgc tgcgctcggt cgttcgggta aagcctgggg
     2401 tgcctaatga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2461 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2521 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2581 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2641 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2701 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2761 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2821 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2881 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     2941 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3001 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3061 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3121 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3181 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     3241 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     3301 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     3361 gcgagaacca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     3421 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     3481 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     3541 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     3601 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     3661 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3721 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     3781 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     3841 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3901 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3961 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     4021 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     4081 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     4141 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     4201 aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     4261 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     4321 gaatagaccg agatagggtt gagtggccgc tacagggcgc tcccattcgc cattcaggct
     4381 gcgcaactgt tgggaagggc gtttcggtgc gggcctcttc gctattacgc cagctggcga
     4441 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     4501 gttgtaaaac gacggccagt gagcgcgacg taatacgact cactataggg cgaattggcg
     4561 gaaggccgtc aaggccgtga cgttaaaact attaagccat ccaatcgacc gttagtcgaa
     4621 tcaggaccgc tggtgcgaga agccgcgaag tatggcgaat gcatcgtata acgtgtggag
     4681 tccgctcatt agagcgtcat gtttagacaa gaaagctaca tatttaattg atcccgatga
     4741 ttttattgat aaattgaccc taactccata cacggtattc tacaatggcg gggttttggt
     4801 caaaatttcc ggactgcgat tgtacatgct gttaacggct ccgcccacta ttaatgaaat
     4861 taaaaattcc aattttaaaa aacgcagcaa gagaaacatt tgtatgaaag aatgcgtaga
     4921 aggaaagaaa aatgtcgtgg acatgctgaa caacaagatt aatatgcctc cgtgtataaa
     4981 aaaaatattg aacgatttga aagaaaacaa tgtaccgcgc ggcggtatgt acaggaagag
     5041 gtttatacta aactgttaca ttgcaaacgt ggtttcgtgt gccaagtgtg aaaaccgatg
     5101 tttaatcaag gctctgacgc atttctacaa ccacgactcc aagtgtgtgg gtgaagtcat
     5161 gcatctttta atcaaatccc aagatgtgta taaaccacca aactgccaaa aaatgaaaac
     5221 tgtggacaag ctctgtccgt ttgctggcaa ctgcaagggt ctcaatccta tttgtaatta
     5281 ttgaataata aaacaattat aaatgctaaa tttgtttttt attaacgata caaaccaaac
     5341 gcaacaagaa catttgtagt attatctata attgaaaacg cgtagttata atcgctgagg
     5401 taatatttaa aatcattttc aaatgattca cagttaattt gcgacaatat aattttattt
     5461 tcacataaac tagacgcctt gtcgtcttct tcttcgtatt ccttctcttt ttcatttttc
     5521 tcctcataaa aattaacata gttattatcg tatccatata tgtatctatc gtatagagta
     5581 aattttttgt tgtcataaat atatatgtct tttttaatgg ggtgtatagt accgctgcgc
     5641 atagtttttc tgtaatttac aacagtgcta ttttctggta gttcttcgga gtgtgttgct
     5701 ttaattatta aatttatata atcaatgaat ttgggatcgt cggttttgta caatatgttg
     5761 ccggcatagt acgcagcttc ttctagttca attacaccat tttttagcag caccggatta
     5821 acataacttt ccaaaatgtt gtacgaaccg ttaaacaaaa acagttcacc tcccttttct
     5881 atactattgt ctgcgagcag ttgtttgttg ttaaaaataa cagccattgt aatgagacgc
     5941 acaaactaat atcacaagct gccaccattt tgttcgccca ggactctagc tatagttcta
     6001 gtggttggct acgtatactc cggaatatta atagatcatg gagataatta aaatgataac
     6061 catctcgcaa ataaataagt attttactgt tttcgtaaca gttttgtaat aaaaaaacct
     6121 ataaat
//
