LOCUS       sequence-232318-        6141 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 N-terminal mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6141)
  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 6141)
  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: pOCC207
FEATURES             Location/Qualifiers
     source          1..6141
                     /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             114..821
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(259..277)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     527..546
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             960..1265
                     /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     1144..1163
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    1438..1572
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1475..1494)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1529..1548
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1645..2017
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(2369..2386)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2479..3067)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2559..2578)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3238..4029,4030..4098))
                     /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     3861..3880
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4099..4203)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4504..4526
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4518..4535
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4519..4535
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4546..4565
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4546..4564
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5175..5299
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5337..5942)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     promoter        6050..6141
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     6066..6083
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
ORIGIN
        1 attccggatt attcataccg tcccaccatc gggcgcggat cgccaccatg ggcagcagcc
       61 atcaccatca ccatcacagc agcggcagac tggaagttct gttccagggg ccgatggtga
      121 gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca
      181 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
      241 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct
      301 gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg
      361 acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
      421 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt
      481 tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga
      541 agaagacgat gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga
      601 agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca
      661 agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca
      721 agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg
      781 agggccgcca ctccaccggc ggcatggacg agctgtacaa gggatccgct ggctccgctg
      841 ctggttctgg cgcggccgca agcgctatgg ccacaacctg ttacattgca caagataaaa
      901 atatatcatc acgaacagta aaactgtctg cttacataaa cagtaataca aggggtgtta
      961 tgcagtttaa ggtttacacc tataaaagag agagccgtta tcgtctgttt gtggatgtac
     1021 agagtgatat tattgacacg cccgggcgac ggatggtgat ccccctggcc agtgcccgtc
     1081 tgctgtcaga taaagtctcc cgtgaacttt acccggtggt gcatatcggg gatgaaagct
     1141 ggcgcatgat gaccaccgat atggccagtg tgccggtctc cgttatcggg gaagaagtgg
     1201 ctgatctcag ccaccgcgaa aatgacatca aaaacgccat taacctgatg ttctggggaa
     1261 tataaatgtc aggctccctt atacacagcc agtggcgcgc cgtaatagaa gcttgtcgag
     1321 aagtactaga ggatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     1381 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     1441 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     1501 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     1561 catgtctgga tctgatcact gcttgagcct aggagatccg aaccagataa gtgaaagcct
     1621 gcatccgcag atcttaatta ataaaacacg atacattgtt attagtacat ttattaagcg
     1681 ctagattctg tgcgttgttg atttacagac aattgttgta cgtattttaa taattcatta
     1741 aatttataat ctttagggtg gtatgttaga gcgaaaatca aatgattttc agcgtcttta
     1801 tatctgaatt taaatattaa atcctcaata gatttgtaaa ataggtttcg attagtttca
     1861 aacaagggtt gtttttccga accgatggct ggactatcta atggattttc gctcaacgcc
     1921 acaaaacttg ccaaatcttg tagcagcaat ctagctttgt cgatattcgt ttgtgttttg
     1981 ttttgtaata aaggttcgac gtcgttcaaa atattatgcg cttttgtatt tctttcatca
     2041 ctgtcgttag tgtacaattg actcgacgta aacacgttaa ataacgcttg gacatattta
     2101 acatcgggcg tgttagcttt attaggccga ttatcgtcgt cgtcccaacc ctcgtcgtta
     2161 gaagttgctt ccgaagacga ttttgccata gccacacgac gcctattaat tgtgtcggct
     2221 aacacgtccg cgatcaaatt tgtagttgag ctttttgggc ctcatgggcc ttccgctcac
     2281 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaacatggt catagctgtt
     2341 tccttgcgta ttgggcgctc tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     2401 gggtaaagcc tggggtgcct aatgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2461 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2521 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2581 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2641 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     2701 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2761 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2821 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     2881 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     2941 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3001 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3061 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3121 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3181 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3241 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3301 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3361 tgctgcaatg ataccgcgag aaccacgctc accggctcca gatttatcag caataaacca
     3421 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3481 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3541 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     3601 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     3661 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     3721 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     3781 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     3841 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     3901 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     3961 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4021 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4081 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4141 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4201 gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc
     4261 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     4321 ccttataaat caaaagaata gaccgagata gggttgagtg gccgctacag ggcgctccca
     4381 ttcgccattc aggctgcgca actgttggga agggcgtttc ggtgcgggcc tcttcgctat
     4441 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     4501 tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgacgtaata cgactcacta
     4561 tagggcgaat tggcggaagg ccgtcaaggc cgtgacgtta aaactattaa gccatccaat
     4621 cgaccgttag tcgaatcagg accgctggtg cgagaagccg cgaagtatgg cgaatgcatc
     4681 gtataacgtg tggagtccgc tcattagagc gtcatgttta gacaagaaag ctacatattt
     4741 aattgatccc gatgatttta ttgataaatt gaccctaact ccatacacgg tattctacaa
     4801 tggcggggtt ttggtcaaaa tttccggact gcgattgtac atgctgttaa cggctccgcc
     4861 cactattaat gaaattaaaa attccaattt taaaaaacgc agcaagagaa acatttgtat
     4921 gaaagaatgc gtagaaggaa agaaaaatgt cgtggacatg ctgaacaaca agattaatat
     4981 gcctccgtgt ataaaaaaaa tattgaacga tttgaaagaa aacaatgtac cgcgcggcgg
     5041 tatgtacagg aagaggttta tactaaactg ttacattgca aacgtggttt cgtgtgccaa
     5101 gtgtgaaaac cgatgtttaa tcaaggctct gacgcatttc tacaaccacg actccaagtg
     5161 tgtgggtgaa gtcatgcatc ttttaatcaa atcccaagat gtgtataaac caccaaactg
     5221 ccaaaaaatg aaaactgtgg acaagctctg tccgtttgct ggcaactgca agggtctcaa
     5281 tcctatttgt aattattgaa taataaaaca attataaatg ctaaatttgt tttttattaa
     5341 cgatacaaac caaacgcaac aagaacattt gtagtattat ctataattga aaacgcgtag
     5401 ttataatcgc tgaggtaata tttaaaatca ttttcaaatg attcacagtt aatttgcgac
     5461 aatataattt tattttcaca taaactagac gccttgtcgt cttcttcttc gtattccttc
     5521 tctttttcat ttttctcctc ataaaaatta acatagttat tatcgtatcc atatatgtat
     5581 ctatcgtata gagtaaattt tttgttgtca taaatatata tgtctttttt aatggggtgt
     5641 atagtaccgc tgcgcatagt ttttctgtaa tttacaacag tgctattttc tggtagttct
     5701 tcggagtgtg ttgctttaat tattaaattt atataatcaa tgaatttggg atcgtcggtt
     5761 ttgtacaata tgttgccggc atagtacgca gcttcttcta gttcaattac accatttttt
     5821 agcagcaccg gattaacata actttccaaa atgttgtacg aaccgttaaa caaaaacagt
     5881 tcacctccct tttctatact attgtctgcg agcagttgtt tgttgttaaa aataacagcc
     5941 attgtaatga gacgcacaaa ctaatatcac aagctgccac cattttgttc gcccaggact
     6001 ctagctatag ttctagtggt tggctacgta tactccggaa tattaataga tcatggagat
     6061 aattaaaatg ataaccatct cgcaaataaa taagtatttt actgttttcg taacagtttt
     6121 gtaataaaaa aacctataaa t
//
