LOCUS       sequence-224659-        6142 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 6142)
  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 6142)
  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..6142
                     /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"
     CDS             230..247
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             260..283
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             284..991
                     /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(429..447)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     697..716
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1130..1435
                     /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     1314..1333
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    1608..1742
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1645..1664)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1699..1718
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1815..2187
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(2539..2556)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2649..3237)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2729..2748)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3408..4199,4200..4268))
                     /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     4031..4050
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4270..4374)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4675..4697
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4689..4706
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4690..4706
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4717..4736
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4717..4735
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5346..5470
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5508..6113)
                     /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 ggcagcagcc atcaccatca
      241 ccatcacagc agcggcagac tggaagttct gttccagggg ccgatggtga gcaagggcga
      301 ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc
      361 cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
      421 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct
      481 gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga
      541 ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
      601 cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa
      661 ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagacgat
      721 gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat
      781 caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta
      841 caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat
      901 cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca
      961 ctccaccggc ggcatggacg agctgtacaa gggatccgct ggctccgctg ctggttctgg
     1021 cgcggccgca agcgctatgg ccacaacctg ttacattgca caagataaaa atatatcatc
     1081 acgaacagta aaactgtctg cttacataaa cagtaataca aggggtgtta tgcagtttaa
     1141 ggtttacacc tataaaagag agagccgtta tcgtctgttt gtggatgtac agagtgatat
     1201 tattgacacg cccgggcgac ggatggtgat ccccctggcc agtgcccgtc tgctgtcaga
     1261 taaagtctcc cgtgaacttt acccggtggt gcatatcggg gatgaaagct ggcgcatgat
     1321 gaccaccgat atggccagtg tgccggtctc cgttatcggg gaagaagtgg ctgatctcag
     1381 ccaccgcgaa aatgacatca aaaacgccat taacctgatg ttctggggaa tataaatgtc
     1441 aggctccctt atacacagcc agtggcgcgc cgtaatagaa gcttgtcgag aagtactaga
     1501 ggatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     1561 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     1621 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     1681 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     1741 tctgatcact gcttgagcct aggagatccg aaccagataa gtgaaagcct gcatccgcag
     1801 atcttaatta ataaaacacg atacattgtt attagtacat ttattaagcg ctagattctg
     1861 tgcgttgttg atttacagac aattgttgta cgtattttaa taattcatta aatttataat
     1921 ctttagggtg gtatgttaga gcgaaaatca aatgattttc agcgtcttta tatctgaatt
     1981 taaatattaa atcctcaata gatttgtaaa ataggtttcg attagtttca aacaagggtt
     2041 gtttttccga accgatggct ggactatcta atggattttc gctcaacgcc acaaaacttg
     2101 ccaaatcttg tagcagcaat ctagctttgt cgatattcgt ttgtgttttg ttttgtaata
     2161 aaggttcgac gtcgttcaaa atattatgcg cttttgtatt tctttcatca ctgtcgttag
     2221 tgtacaattg actcgacgta aacacgttaa ataacgcttg gacatattta acatcgggcg
     2281 tgttagcttt attaggccga ttatcgtcgt cgtcccaacc ctcgtcgtta gaagttgctt
     2341 ccgaagacga ttttgccata gccacacgac gcctattaat tgtgtcggct aacacgtccg
     2401 cgatcaaatt tgtagttgag ctttttgggc ctcatgggcc ttccgctcac tgcccgcttt
     2461 ccagtcggga aacctgtcgt gccagctgca ttaacatggt catagctgtt tccttgcgta
     2521 ttgggcgctc tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc gggtaaagcc
     2581 tggggtgcct aatgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     2641 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2701 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2761 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     2821 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     2881 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     2941 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3001 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3061 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     3121 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3181 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3241 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3301 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3361 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     3421 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     3481 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     3541 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     3601 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     3661 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     3721 gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     3781 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     3841 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     3901 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     3961 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     4021 tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     4081 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     4141 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     4201 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     4261 atactcatta ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     4321 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     4381 tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     4441 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     4501 tcaaaagaat agaccgagat agggttgagt ggccgctaca gggcgctccc attcgccatt
     4561 caggctgcgc aactgttggg aagggcgttt cggtgcgggc ctcttcgcta ttacgccagc
     4621 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
     4681 cacgacgttg taaaacgacg gccagtgagc gcgacgtaat acgactcact atagggcgaa
     4741 ttggcggaag gccgtcaagg ccgtgacgtt aaaactatta agccatccaa tcgaccgtta
     4801 gtcgaatcag gaccgctggt gcgagaagcc gcgaagtatg gcgaatgcat cgtataacgt
     4861 gtggagtccg ctcattagag cgtcatgttt agacaagaaa gctacatatt taattgatcc
     4921 cgatgatttt attgataaat tgaccctaac tccatacacg gtattctaca atggcggggt
     4981 tttggtcaaa atttccggac tgcgattgta catgctgtta acggctccgc ccactattaa
     5041 tgaaattaaa aattccaatt ttaaaaaacg cagcaagaga aacatttgta tgaaagaatg
     5101 cgtagaagga aagaaaaatg tcgtggacat gctgaacaac aagattaata tgcctccgtg
     5161 tataaaaaaa atattgaacg atttgaaaga aaacaatgta ccgcgcggcg gtatgtacag
     5221 gaagaggttt atactaaact gttacattgc aaacgtggtt tcgtgtgcca agtgtgaaaa
     5281 ccgatgttta atcaaggctc tgacgcattt ctacaaccac gactccaagt gtgtgggtga
     5341 agtcatgcat cttttaatca aatcccaaga tgtgtataaa ccaccaaact gccaaaaaat
     5401 gaaaactgtg gacaagctct gtccgtttgc tggcaactgc aagggtctca atcctatttg
     5461 taattattga ataataaaac aattataaat gctaaatttg ttttttatta acgatacaaa
     5521 ccaaacgcaa caagaacatt tgtagtatta tctataattg aaaacgcgta gttataatcg
     5581 ctgaggtaat atttaaaatc attttcaaat gattcacagt taatttgcga caatataatt
     5641 ttattttcac ataaactaga cgccttgtcg tcttcttctt cgtattcctt ctctttttca
     5701 tttttctcct cataaaaatt aacatagtta ttatcgtatc catatatgta tctatcgtat
     5761 agagtaaatt ttttgttgtc ataaatatat atgtcttttt taatggggtg tatagtaccg
     5821 ctgcgcatag tttttctgta atttacaaca gtgctatttt ctggtagttc ttcggagtgt
     5881 gttgctttaa ttattaaatt tatataatca atgaatttgg gatcgtcggt tttgtacaat
     5941 atgttgccgg catagtacgc agcttcttct agttcaatta caccattttt tagcagcacc
     6001 ggattaacat aactttccaa aatgttgtac gaaccgttaa acaaaaacag ttcacctccc
     6061 ttttctatac tattgtctgc gagcagttgt ttgttgttaa aaataacagc cattgtaatg
     6121 agacgcacaa actaatatca ca
//
