LOCUS       sequence-232327-        5421 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with N-terminal HIS6, cleavable with 3C, and
            C-terminal sortase site (LPETGg).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5421)
  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 5421)
  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: pOCC110
FEATURES             Location/Qualifiers
     source          1..5421
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..92
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     17..34
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     regulatory      134..143
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             152..169
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             182..205
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             314..619
                     /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     498..517
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    810..944
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(847..866)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     901..920
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1017..1389
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(1741..1758)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1851..2439)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1931..1950)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2610..3401,3402..3470))
                     /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     3233..3252
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3471..3575)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3876..3898
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3890..3907
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3891..3907
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3918..3937
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3918..3936
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4547..4671
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(4709..5314)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
ORIGIN
        1 atcatggaga taattaaaat gataaccatc tcgcaaataa ataagtattt tactgttttc
       61 gtaacagttt tgtaataaaa aaacctataa atattccgga ttattcatac cgtcccacca
      121 tcgggcgcgg atcgccacca tgggcagcag ccatcaccat caccatcaca gcagcggcag
      181 actggaagtt ctgttccagg ggccggcggc cgcaagcgct atggccacaa cctgttacat
      241 tgcacaagat aaaaatatat catcacgaac agtaaaactg tctgcttaca taaacagtaa
      301 tacaaggggt gttatgcagt ttaaggttta cacctataaa agagagagcc gttatcgtct
      361 gtttgtggat gtacagagtg atattattga cacgcccggg cgacggatgg tgatccccct
      421 ggccagtgcc cgtctgctgt cagataaagt ctcccgtgaa ctttacccgg tggtgcatat
      481 cggggatgaa agctggcgca tgatgaccac cgatatggcc agtgtgccgg tctccgttat
      541 cggggaagaa gtggctgatc tcagccaccg cgaaaatgac atcaaaaacg ccattaacct
      601 gatgttctgg ggaatataaa tgtcaggctc ccttatacac agccagtggc gcgccgctgc
      661 ccgagaccgg cggctaatag aagcttgtcg agaagtacta gaggatcata atcagccata
      721 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
      781 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
      841 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
      901 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatctgatca ctgcttgagc
      961 ctaggagatc cgaaccagat aagtgaaagc ctgcatccgc agatcttaat taataaaaca
     1021 cgatacattg ttattagtac atttattaag cgctagattc tgtgcgttgt tgatttacag
     1081 acaattgttg tacgtatttt aataattcat taaatttata atctttaggg tggtatgtta
     1141 gagcgaaaat caaatgattt tcagcgtctt tatatctgaa tttaaatatt aaatcctcaa
     1201 tagatttgta aaataggttt cgattagttt caaacaaggg ttgtttttcc gaaccgatgg
     1261 ctggactatc taatggattt tcgctcaacg ccacaaaact tgccaaatct tgtagcagca
     1321 atctagcttt gtcgatattc gtttgtgttt tgttttgtaa taaaggttcg acgtcgttca
     1381 aaatattatg cgcttttgta tttctttcat cactgtcgtt agtgtacaat tgactcgacg
     1441 taaacacgtt aaataacgct tggacatatt taacatcggg cgtgttagct ttattaggcc
     1501 gattatcgtc gtcgtcccaa ccctcgtcgt tagaagttgc ttccgaagac gattttgcca
     1561 tagccacacg acgcctatta attgtgtcgg ctaacacgtc cgcgatcaaa tttgtagttg
     1621 agctttttgg gcctcatggg ccttccgctc actgcccgct ttccagtcgg gaaacctgtc
     1681 gtgccagctg cattaacatg gtcatagctg tttccttgcg tattgggcgc tctccgcttc
     1741 ctcgctcact gactcgctgc gctcggtcgt tcgggtaaag cctggggtgc ctaatgagca
     1801 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     1861 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     1921 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     1981 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2041 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2101 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2161 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2221 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     2281 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     2341 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     2401 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     2461 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     2521 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     2581 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     2641 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     2701 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agaaccacgc
     2761 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     2821 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     2881 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     2941 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     3001 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     3061 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     3121 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     3181 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     3241 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     3301 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     3361 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     3421 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     3481 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     3541 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     3601 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     3661 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     3721 tagggttgag tggccgctac agggcgctcc cattcgccat tcaggctgcg caactgttgg
     3781 gaagggcgtt tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     3841 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     3901 ggccagtgag cgcgacgtaa tacgactcac tatagggcga attggcggaa ggccgtcaag
     3961 gccgtgacgt taaaactatt aagccatcca atcgaccgtt agtcgaatca ggaccgctgg
     4021 tgcgagaagc cgcgaagtat ggcgaatgca tcgtataacg tgtggagtcc gctcattaga
     4081 gcgtcatgtt tagacaagaa agctacatat ttaattgatc ccgatgattt tattgataaa
     4141 ttgaccctaa ctccatacac ggtattctac aatggcgggg ttttggtcaa aatttccgga
     4201 ctgcgattgt acatgctgtt aacggctccg cccactatta atgaaattaa aaattccaat
     4261 tttaaaaaac gcagcaagag aaacatttgt atgaaagaat gcgtagaagg aaagaaaaat
     4321 gtcgtggaca tgctgaacaa caagattaat atgcctccgt gtataaaaaa aatattgaac
     4381 gatttgaaag aaaacaatgt accgcgcggc ggtatgtaca ggaagaggtt tatactaaac
     4441 tgttacattg caaacgtggt ttcgtgtgcc aagtgtgaaa accgatgttt aatcaaggct
     4501 ctgacgcatt tctacaacca cgactccaag tgtgtgggtg aagtcatgca tcttttaatc
     4561 aaatcccaag atgtgtataa accaccaaac tgccaaaaaa tgaaaactgt ggacaagctc
     4621 tgtccgtttg ctggcaactg caagggtctc aatcctattt gtaattattg aataataaaa
     4681 caattataaa tgctaaattt gttttttatt aacgatacaa accaaacgca acaagaacat
     4741 ttgtagtatt atctataatt gaaaacgcgt agttataatc gctgaggtaa tatttaaaat
     4801 cattttcaaa tgattcacag ttaatttgcg acaatataat tttattttca cataaactag
     4861 acgccttgtc gtcttcttct tcgtattcct tctctttttc atttttctcc tcataaaaat
     4921 taacatagtt attatcgtat ccatatatgt atctatcgta tagagtaaat tttttgttgt
     4981 cataaatata tatgtctttt ttaatggggt gtatagtacc gctgcgcata gtttttctgt
     5041 aatttacaac agtgctattt tctggtagtt cttcggagtg tgttgcttta attattaaat
     5101 ttatataatc aatgaatttg ggatcgtcgg ttttgtacaa tatgttgccg gcatagtacg
     5161 cagcttcttc tagttcaatt acaccatttt ttagcagcac cggattaaca taactttcca
     5221 aaatgttgta cgaaccgtta aacaaaaaca gttcacctcc cttttctata ctattgtctg
     5281 cgagcagttg tttgttgtta aaaataacag ccattgtaat gagacgcaca aactaatatc
     5341 acaagctgcc accattttgt tcgcccagga ctctagctat agttctagtg gttggctacg
     5401 tatactccgg aatattaata g
//
