LOCUS       sequence-224669-        6118 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with C-terminal Dendra2, and C-terminal HIS6
            tag, cleavable with 3C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6118)
  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 6118)
  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: pOCC215
FEATURES             Location/Qualifiers
     source          1..6118
                     /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..1391
                     /codon_start=1
                     /product="monomeric photoswitchable green-to-red 
                     fluorescent protein from Dendronephthya"
                     /label=Dendra2
                     /note="mammalian codon-optimized"
                     /translation="MNTPGINLIKEDMRVKVHMEGNVNGHAFVIEGEGKGKPYEGTQTA
                     NLTVKEGAPLPFSYDILTTAVHYGNRVFTKYPEDIPDYFKQSFPEGYSWERTMTFEDKG
                     ICTIRSDISLEGDCFFQNVRFKGTNFPPNGPVMQKKTLKWEPSTEKLHVRDGLLVGNIN
                     MALLLEGGGHYLCDFKTTYKAKKVVQLPDAHFVDHRIEILGNDSDYNKVKLYEHAVARY
                     SPLPSQVW"
     CDS             1392..1415
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1425..1442
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1584..1718
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1621..1640)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1675..1694
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1791..2163
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(2515..2532)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2625..3213)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2705..2724)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3384..4175,4176..4244))
                     /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     4007..4026
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4246..4350)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4651..4673
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4665..4682
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4666..4682
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4693..4712
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4693..4711
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5322..5446
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5484..6089)
                     /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 catgaacacc ccgggaatta
      721 acctgatcaa ggaggacatg cgcgtgaagg tgcacatgga gggcaacgtg aacggccacg
      781 ccttcgtgat cgagggcgag ggcaagggca agccctacga gggcacccag accgccaacc
      841 tgaccgtgaa ggagggcgcc cccctgccct tcagctacga catcctgacc accgccgtgc
      901 actacggcaa ccgggtgttc accaagtacc ccgaggacat ccccgactac ttcaagcaga
      961 gcttccccga gggctacagc tgggagcgca ccatgacctt cgaggacaag ggcatctgca
     1021 ccatccgcag cgacatcagc ctggagggcg actgcttctt ccagaacgtg cgcttcaagg
     1081 gcaccaactt cccccccaac ggccccgtga tgcagaagaa gaccctgaag tgggagccca
     1141 gcaccgagaa gctgcacgtg cgcgacggcc tgctggtggg caacatcaac atggccctgc
     1201 tgctggaggg cggcggccac tacctgtgcg acttcaagac cacctacaag gccaagaagg
     1261 tggtgcagct gcccgacgcc cacttcgtgg accaccgcat cgagatcctg ggcaacgaca
     1321 gcgactacaa caaggtgaag ctgtacgagc acgccgtggc ccgctacagc cccctgccca
     1381 gccaggtgtg gctggaagtt ctgttccagg ggccgggcag cagccatcac catcaccatc
     1441 acagtggata atagaagctt gtcgagaagt actagaggat cataatcagc cataccacat
     1501 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     1561 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     1621 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     1681 tgtccaaact catcaatgta tcttatcatg tctggatctg atcactgctt gagcctagga
     1741 gatccgaacc agataagtga aagcctgcat ccgcagatct taattaataa aacacgatac
     1801 attgttatta gtacatttat taagcgctag attctgtgcg ttgttgattt acagacaatt
     1861 gttgtacgta ttttaataat tcattaaatt tataatcttt agggtggtat gttagagcga
     1921 aaatcaaatg attttcagcg tctttatatc tgaatttaaa tattaaatcc tcaatagatt
     1981 tgtaaaatag gtttcgatta gtttcaaaca agggttgttt ttccgaaccg atggctggac
     2041 tatctaatgg attttcgctc aacgccacaa aacttgccaa atcttgtagc agcaatctag
     2101 ctttgtcgat attcgtttgt gttttgtttt gtaataaagg ttcgacgtcg ttcaaaatat
     2161 tatgcgcttt tgtatttctt tcatcactgt cgttagtgta caattgactc gacgtaaaca
     2221 cgttaaataa cgcttggaca tatttaacat cgggcgtgtt agctttatta ggccgattat
     2281 cgtcgtcgtc ccaaccctcg tcgttagaag ttgcttccga agacgatttt gccatagcca
     2341 cacgacgcct attaattgtg tcggctaaca cgtccgcgat caaatttgta gttgagcttt
     2401 ttgggcctca tgggccttcc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     2461 gctgcattaa catggtcata gctgtttcct tgcgtattgg gcgctctccg cttcctcgct
     2521 cactgactcg ctgcgctcgg tcgttcgggt aaagcctggg gtgcctaatg agcaaaaggc
     2581 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2641 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2701 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2761 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2821 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2881 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2941 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3001 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3061 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3121 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3181 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     3241 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     3301 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     3361 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     3421 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     3481 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     3541 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     3601 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3661 tcgccagtta atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc
     3721 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3781 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     3841 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     3901 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     3961 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca
     4021 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     4081 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     4141 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     4201 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcattactct tcctttttca
     4261 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4321 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt
     4381 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     4441 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     4501 ttgagtggcc gctacagggc gctcccattc gccattcagg ctgcgcaact gttgggaagg
     4561 gcgtttcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     4621 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     4681 gtgagcgcga cgtaatacga ctcactatag ggcgaattgg cggaaggccg tcaaggccgt
     4741 gacgttaaaa ctattaagcc atccaatcga ccgttagtcg aatcaggacc gctggtgcga
     4801 gaagccgcga agtatggcga atgcatcgta taacgtgtgg agtccgctca ttagagcgtc
     4861 atgtttagac aagaaagcta catatttaat tgatcccgat gattttattg ataaattgac
     4921 cctaactcca tacacggtat tctacaatgg cggggttttg gtcaaaattt ccggactgcg
     4981 attgtacatg ctgttaacgg ctccgcccac tattaatgaa attaaaaatt ccaattttaa
     5041 aaaacgcagc aagagaaaca tttgtatgaa agaatgcgta gaaggaaaga aaaatgtcgt
     5101 ggacatgctg aacaacaaga ttaatatgcc tccgtgtata aaaaaaatat tgaacgattt
     5161 gaaagaaaac aatgtaccgc gcggcggtat gtacaggaag aggtttatac taaactgtta
     5221 cattgcaaac gtggtttcgt gtgccaagtg tgaaaaccga tgtttaatca aggctctgac
     5281 gcatttctac aaccacgact ccaagtgtgt gggtgaagtc atgcatcttt taatcaaatc
     5341 ccaagatgtg tataaaccac caaactgcca aaaaatgaaa actgtggaca agctctgtcc
     5401 gtttgctggc aactgcaagg gtctcaatcc tatttgtaat tattgaataa taaaacaatt
     5461 ataaatgcta aatttgtttt ttattaacga tacaaaccaa acgcaacaag aacatttgta
     5521 gtattatcta taattgaaaa cgcgtagtta taatcgctga ggtaatattt aaaatcattt
     5581 tcaaatgatt cacagttaat ttgcgacaat ataattttat tttcacataa actagacgcc
     5641 ttgtcgtctt cttcttcgta ttccttctct ttttcatttt tctcctcata aaaattaaca
     5701 tagttattat cgtatccata tatgtatcta tcgtatagag taaatttttt gttgtcataa
     5761 atatatatgt cttttttaat ggggtgtata gtaccgctgc gcatagtttt tctgtaattt
     5821 acaacagtgc tattttctgg tagttcttcg gagtgtgttg ctttaattat taaatttata
     5881 taatcaatga atttgggatc gtcggttttg tacaatatgt tgccggcata gtacgcagct
     5941 tcttctagtt caattacacc attttttagc agcaccggat taacataact ttccaaaatg
     6001 ttgtacgaac cgttaaacaa aaacagttca cctccctttt ctatactatt gtctgcgagc
     6061 agttgtttgt tgttaaaaat aacagccatt gtaatgagac gcacaaacta atatcaca
//
