LOCUS       sequence-232276-        5403 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 
            protease.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5403)
  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 5403)
  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: pOCC5
FEATURES             Location/Qualifiers
     source          1..5403
                     /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"
     polyA_signal    700..834
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(737..756)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     791..810
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           907..1279
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(1631..1648)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1741..2329)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1821..1840)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2500..3291,3292..3360))
                     /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     3123..3142
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3361..3465)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3766..3788
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3780..3797
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3781..3797
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3808..3827
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3808..3826
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4437..4561
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(4599..5204)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     promoter        5312..5403
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     5328..5345
                     /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 gccgtaatag aagcttgtcg agaagtacta gaggatcata
      601 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
      661 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
      721 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
      781 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatctgatca
      841 ctgcttgagc ctaggagatc cgaaccagat aagtgaaagc ctgcatccgc agatcttaat
      901 taataaaaca cgatacattg ttattagtac atttattaag cgctagattc tgtgcgttgt
      961 tgatttacag acaattgttg tacgtatttt aataattcat taaatttata atctttaggg
     1021 tggtatgtta gagcgaaaat caaatgattt tcagcgtctt tatatctgaa tttaaatatt
     1081 aaatcctcaa tagatttgta aaataggttt cgattagttt caaacaaggg ttgtttttcc
     1141 gaaccgatgg ctggactatc taatggattt tcgctcaacg ccacaaaact tgccaaatct
     1201 tgtagcagca atctagcttt gtcgatattc gtttgtgttt tgttttgtaa taaaggttcg
     1261 acgtcgttca aaatattatg cgcttttgta tttctttcat cactgtcgtt agtgtacaat
     1321 tgactcgacg taaacacgtt aaataacgct tggacatatt taacatcggg cgtgttagct
     1381 ttattaggcc gattatcgtc gtcgtcccaa ccctcgtcgt tagaagttgc ttccgaagac
     1441 gattttgcca tagccacacg acgcctatta attgtgtcgg ctaacacgtc cgcgatcaaa
     1501 tttgtagttg agctttttgg gcctcatggg ccttccgctc actgcccgct ttccagtcgg
     1561 gaaacctgtc gtgccagctg cattaacatg gtcatagctg tttccttgcg tattgggcgc
     1621 tctccgcttc ctcgctcact gactcgctgc gctcggtcgt tcgggtaaag cctggggtgc
     1681 ctaatgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     1741 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     1801 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     1861 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     1921 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     1981 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     2041 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     2101 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     2161 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     2221 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     2281 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     2341 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     2401 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     2461 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     2521 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     2581 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     2641 agaaccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     2701 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     2761 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     2821 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     2881 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     2941 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     3001 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     3061 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     3121 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     3181 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     3241 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     3301 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     3361 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     3421 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     3481 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     3541 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     3601 tagaccgaga tagggttgag tggccgctac agggcgctcc cattcgccat tcaggctgcg
     3661 caactgttgg gaagggcgtt tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     3721 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     3781 gtaaaacgac ggccagtgag cgcgacgtaa tacgactcac tatagggcga attggcggaa
     3841 ggccgtcaag gccgtgacgt taaaactatt aagccatcca atcgaccgtt agtcgaatca
     3901 ggaccgctgg tgcgagaagc cgcgaagtat ggcgaatgca tcgtataacg tgtggagtcc
     3961 gctcattaga gcgtcatgtt tagacaagaa agctacatat ttaattgatc ccgatgattt
     4021 tattgataaa ttgaccctaa ctccatacac ggtattctac aatggcgggg ttttggtcaa
     4081 aatttccgga ctgcgattgt acatgctgtt aacggctccg cccactatta atgaaattaa
     4141 aaattccaat tttaaaaaac gcagcaagag aaacatttgt atgaaagaat gcgtagaagg
     4201 aaagaaaaat gtcgtggaca tgctgaacaa caagattaat atgcctccgt gtataaaaaa
     4261 aatattgaac gatttgaaag aaaacaatgt accgcgcggc ggtatgtaca ggaagaggtt
     4321 tatactaaac tgttacattg caaacgtggt ttcgtgtgcc aagtgtgaaa accgatgttt
     4381 aatcaaggct ctgacgcatt tctacaacca cgactccaag tgtgtgggtg aagtcatgca
     4441 tcttttaatc aaatcccaag atgtgtataa accaccaaac tgccaaaaaa tgaaaactgt
     4501 ggacaagctc tgtccgtttg ctggcaactg caagggtctc aatcctattt gtaattattg
     4561 aataataaaa caattataaa tgctaaattt gttttttatt aacgatacaa accaaacgca
     4621 acaagaacat ttgtagtatt atctataatt gaaaacgcgt agttataatc gctgaggtaa
     4681 tatttaaaat cattttcaaa tgattcacag ttaatttgcg acaatataat tttattttca
     4741 cataaactag acgccttgtc gtcttcttct tcgtattcct tctctttttc atttttctcc
     4801 tcataaaaat taacatagtt attatcgtat ccatatatgt atctatcgta tagagtaaat
     4861 tttttgttgt cataaatata tatgtctttt ttaatggggt gtatagtacc gctgcgcata
     4921 gtttttctgt aatttacaac agtgctattt tctggtagtt cttcggagtg tgttgcttta
     4981 attattaaat ttatataatc aatgaatttg ggatcgtcgg ttttgtacaa tatgttgccg
     5041 gcatagtacg cagcttcttc tagttcaatt acaccatttt ttagcagcac cggattaaca
     5101 taactttcca aaatgttgta cgaaccgtta aacaaaaaca gttcacctcc cttttctata
     5161 ctattgtctg cgagcagttg tttgttgtta aaaataacag ccattgtaat gagacgcaca
     5221 aactaatatc acaagctgcc accattttgt tcgcccagga ctctagctat agttctagtg
     5281 gttggctacg tatactccgg aatattaata gatcatggag ataattaaaa tgataaccat
     5341 ctcgcaaata aataagtatt ttactgtttt cgtaacagtt ttgtaataaa aaaacctata
     5401 aat
//
