LOCUS       sequence-224670-        6124 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with C-terminal mKate2, and C-terminal HIS6 
            tag, cleavable with 3C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6124)
  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 6124)
  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: pOCC202
FEATURES             Location/Qualifiers
     source          1..6124
                     /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             702..1397
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     CDS             1398..1421
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1431..1448
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1590..1724
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1627..1646)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1681..1700
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1797..2169
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(2521..2538)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2631..3219)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2711..2730)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3390..4181,4182..4250))
                     /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     4013..4032
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4252..4356)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4657..4679
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4671..4688
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4672..4688
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4699..4718
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4699..4717
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5328..5452
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5490..6095)
                     /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 gggatccgct ggctccgctg ctggttctgg catggtgagc gagctgatta
      721 aggagaacat gcacatgaag ctgtacatgg agggcaccgt gaacaaccac cacttcaagt
      781 gcacatccga gggcgaaggc aagccctacg agggcaccca gaccatgaga atcaaggcgg
      841 tcgagggcgg ccctctcccc ttcgccttcg acatcctggc taccagcttc atgtacggca
      901 gcaaaacctt catcaaccac acccagggca tccccgactt ctttaagcag tccttccccg
      961 agggcttcac atgggagaga gtcaccacat acgaagacgg gggcgtgctg accgctaccc
     1021 aggacaccag cctccaggac ggctgcctca tctacaacgt caagatcaga ggggtgaact
     1081 tcccatccaa cggccctgtg atgcagaaga aaacactcgg ctgggaggcc tccaccgaga
     1141 ccctgtaccc cgctgacggc ggcctggaag gcagagccga catggccctg aagctcgtgg
     1201 gcgggggcca cctgatctgc aacttgaaga ccacatacag atccaagaaa cccgctaaga
     1261 acctcaagat gcccggcgtc tactatgtgg acagaagact ggaaagaatc aaggaggccg
     1321 acaaagagac ctacgtcgag cagcacgagg tggctgtggc cagatactgc gacctcccta
     1381 gcaaactggg gcacagactg gaagttctgt tccaggggcc gggcagcagc catcaccatc
     1441 accatcacag tggataatag aagcttgtcg agaagtacta gaggatcata atcagccata
     1501 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     1561 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     1621 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     1681 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatctgatca ctgcttgagc
     1741 ctaggagatc cgaaccagat aagtgaaagc ctgcatccgc agatcttaat taataaaaca
     1801 cgatacattg ttattagtac atttattaag cgctagattc tgtgcgttgt tgatttacag
     1861 acaattgttg tacgtatttt aataattcat taaatttata atctttaggg tggtatgtta
     1921 gagcgaaaat caaatgattt tcagcgtctt tatatctgaa tttaaatatt aaatcctcaa
     1981 tagatttgta aaataggttt cgattagttt caaacaaggg ttgtttttcc gaaccgatgg
     2041 ctggactatc taatggattt tcgctcaacg ccacaaaact tgccaaatct tgtagcagca
     2101 atctagcttt gtcgatattc gtttgtgttt tgttttgtaa taaaggttcg acgtcgttca
     2161 aaatattatg cgcttttgta tttctttcat cactgtcgtt agtgtacaat tgactcgacg
     2221 taaacacgtt aaataacgct tggacatatt taacatcggg cgtgttagct ttattaggcc
     2281 gattatcgtc gtcgtcccaa ccctcgtcgt tagaagttgc ttccgaagac gattttgcca
     2341 tagccacacg acgcctatta attgtgtcgg ctaacacgtc cgcgatcaaa tttgtagttg
     2401 agctttttgg gcctcatggg ccttccgctc actgcccgct ttccagtcgg gaaacctgtc
     2461 gtgccagctg cattaacatg gtcatagctg tttccttgcg tattgggcgc tctccgcttc
     2521 ctcgctcact gactcgctgc gctcggtcgt tcgggtaaag cctggggtgc ctaatgagca
     2581 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     2641 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2701 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2761 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2821 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2881 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2941 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3001 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3061 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3121 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3181 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3241 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     3301 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     3361 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     3421 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     3481 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     3541 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     3601 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     3661 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg
     3721 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     3781 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     3841 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     3901 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     3961 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc
     4021 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     4081 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     4141 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     4201 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat tactcttcct
     4261 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4321 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4381 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     4441 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     4501 atagggttga gtggccgcta cagggcgctc ccattcgcca ttcaggctgc gcaactgttg
     4561 ggaagggcgt ttcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     4621 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     4681 cggccagtga gcgcgacgta atacgactca ctatagggcg aattggcgga aggccgtcaa
     4741 ggccgtgacg ttaaaactat taagccatcc aatcgaccgt tagtcgaatc aggaccgctg
     4801 gtgcgagaag ccgcgaagta tggcgaatgc atcgtataac gtgtggagtc cgctcattag
     4861 agcgtcatgt ttagacaaga aagctacata tttaattgat cccgatgatt ttattgataa
     4921 attgacccta actccataca cggtattcta caatggcggg gttttggtca aaatttccgg
     4981 actgcgattg tacatgctgt taacggctcc gcccactatt aatgaaatta aaaattccaa
     5041 ttttaaaaaa cgcagcaaga gaaacatttg tatgaaagaa tgcgtagaag gaaagaaaaa
     5101 tgtcgtggac atgctgaaca acaagattaa tatgcctccg tgtataaaaa aaatattgaa
     5161 cgatttgaaa gaaaacaatg taccgcgcgg cggtatgtac aggaagaggt ttatactaaa
     5221 ctgttacatt gcaaacgtgg tttcgtgtgc caagtgtgaa aaccgatgtt taatcaaggc
     5281 tctgacgcat ttctacaacc acgactccaa gtgtgtgggt gaagtcatgc atcttttaat
     5341 caaatcccaa gatgtgtata aaccaccaaa ctgccaaaaa atgaaaactg tggacaagct
     5401 ctgtccgttt gctggcaact gcaagggtct caatcctatt tgtaattatt gaataataaa
     5461 acaattataa atgctaaatt tgttttttat taacgataca aaccaaacgc aacaagaaca
     5521 tttgtagtat tatctataat tgaaaacgcg tagttataat cgctgaggta atatttaaaa
     5581 tcattttcaa atgattcaca gttaatttgc gacaatataa ttttattttc acataaacta
     5641 gacgccttgt cgtcttcttc ttcgtattcc ttctcttttt catttttctc ctcataaaaa
     5701 ttaacatagt tattatcgta tccatatatg tatctatcgt atagagtaaa ttttttgttg
     5761 tcataaatat atatgtcttt tttaatgggg tgtatagtac cgctgcgcat agtttttctg
     5821 taatttacaa cagtgctatt ttctggtagt tcttcggagt gtgttgcttt aattattaaa
     5881 tttatataat caatgaattt gggatcgtcg gttttgtaca atatgttgcc ggcatagtac
     5941 gcagcttctt ctagttcaat tacaccattt tttagcagca ccggattaac ataactttcc
     6001 aaaatgttgt acgaaccgtt aaacaaaaac agttcacctc ccttttctat actattgtct
     6061 gcgagcagtt gtttgttgtt aaaaataaca gccattgtaa tgagacgcac aaactaatat
     6121 caca
//
