LOCUS       sequence-232266-        5472 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with honey bee melitine secretion signal and
            C-terminal HIS6, cleavable with 3C protease.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5472)
  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 5472)
  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: pOCC167
FEATURES             Location/Qualifiers
     source          1..5472
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     sig_peptide     1..60
                     /codon_start=1
                     /product="signal sequence from honeybee melittin"
                     /label=melittin signal sequence
                     /translation="KFLVNVALVFMVVYISYIYA"
     CDS             181..486
                     /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     365..384
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             524..547
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             557..574
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    716..850
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(753..772)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     807..826
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           923..1295
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(1647..1664)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1757..2345)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1837..1856)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2516..3307,3308..3376))
                     /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     3139..3158
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3377..3481)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3782..3804
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3796..3813
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3797..3813
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3824..3843
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3824..3842
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4453..4577
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(4615..5220)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     promoter        5328..5419
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     5344..5361
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     regulatory      5461..5470
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 aaattcttag tcaacgttgc ccttgttttt atggtcgtat acatttctta catctatgcg
       61 ggatccaatt gggcggccgc aagcgctatg gccacaacct gttacattgc acaagataaa
      121 aatatatcat cacgaacagt aaaactgtct gcttacataa acagtaatac aaggggtgtt
      181 atgcagttta aggtttacac ctataaaaga gagagccgtt atcgtctgtt tgtggatgta
      241 cagagtgata ttattgacac gcccgggcga cggatggtga tccccctggc cagtgcccgt
      301 ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc
      361 tggcgcatga tgaccaccga tatggccagt gtgccggtct ccgttatcgg ggaagaagtg
      421 gctgatctca gccaccgcga aaatgacatc aaaaacgcca ttaacctgat gttctgggga
      481 atataaatgt caggctccct tatacacagc cagtggcgcg ccgctggaag ttctgttcca
      541 ggggccgggc agcagccatc accatcacca tcacagtgga taatagaagc ttgtcgagaa
      601 gtactagagg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
      661 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
      721 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
      781 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
      841 tgtctggatc tgatcactgc ttgagcctag gagatccgaa ccagataagt gaaagcctgc
      901 atccgcagat cttaattaat aaaacacgat acattgttat tagtacattt attaagcgct
      961 agattctgtg cgttgttgat ttacagacaa ttgttgtacg tattttaata attcattaaa
     1021 tttataatct ttagggtggt atgttagagc gaaaatcaaa tgattttcag cgtctttata
     1081 tctgaattta aatattaaat cctcaataga tttgtaaaat aggtttcgat tagtttcaaa
     1141 caagggttgt ttttccgaac cgatggctgg actatctaat ggattttcgc tcaacgccac
     1201 aaaacttgcc aaatcttgta gcagcaatct agctttgtcg atattcgttt gtgttttgtt
     1261 ttgtaataaa ggttcgacgt cgttcaaaat attatgcgct tttgtatttc tttcatcact
     1321 gtcgttagtg tacaattgac tcgacgtaaa cacgttaaat aacgcttgga catatttaac
     1381 atcgggcgtg ttagctttat taggccgatt atcgtcgtcg tcccaaccct cgtcgttaga
     1441 agttgcttcc gaagacgatt ttgccatagc cacacgacgc ctattaattg tgtcggctaa
     1501 cacgtccgcg atcaaatttg tagttgagct ttttgggcct catgggcctt ccgctcactg
     1561 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aacatggtca tagctgtttc
     1621 cttgcgtatt gggcgctctc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     1681 gtaaagcctg gggtgcctaa tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     1741 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     1801 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     1861 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     1921 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     1981 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     2041 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     2101 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2161 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     2221 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     2281 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     2341 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     2401 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     2461 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     2521 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     2581 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     2641 ctgcaatgat accgcgagaa ccacgctcac cggctccaga tttatcagca ataaaccagc
     2701 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     2761 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     2821 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     2881 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     2941 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     3001 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     3061 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     3121 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     3181 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     3241 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     3301 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     3361 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     3421 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     3481 gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc
     3541 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     3601 ttataaatca aaagaataga ccgagatagg gttgagtggc cgctacaggg cgctcccatt
     3661 cgccattcag gctgcgcaac tgttgggaag ggcgtttcgg tgcgggcctc ttcgctatta
     3721 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     3781 tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg acgtaatacg actcactata
     3841 gggcgaattg gcggaaggcc gtcaaggccg tgacgttaaa actattaagc catccaatcg
     3901 accgttagtc gaatcaggac cgctggtgcg agaagccgcg aagtatggcg aatgcatcgt
     3961 ataacgtgtg gagtccgctc attagagcgt catgtttaga caagaaagct acatatttaa
     4021 ttgatcccga tgattttatt gataaattga ccctaactcc atacacggta ttctacaatg
     4081 gcggggtttt ggtcaaaatt tccggactgc gattgtacat gctgttaacg gctccgccca
     4141 ctattaatga aattaaaaat tccaatttta aaaaacgcag caagagaaac atttgtatga
     4201 aagaatgcgt agaaggaaag aaaaatgtcg tggacatgct gaacaacaag attaatatgc
     4261 ctccgtgtat aaaaaaaata ttgaacgatt tgaaagaaaa caatgtaccg cgcggcggta
     4321 tgtacaggaa gaggtttata ctaaactgtt acattgcaaa cgtggtttcg tgtgccaagt
     4381 gtgaaaaccg atgtttaatc aaggctctga cgcatttcta caaccacgac tccaagtgtg
     4441 tgggtgaagt catgcatctt ttaatcaaat cccaagatgt gtataaacca ccaaactgcc
     4501 aaaaaatgaa aactgtggac aagctctgtc cgtttgctgg caactgcaag ggtctcaatc
     4561 ctatttgtaa ttattgaata ataaaacaat tataaatgct aaatttgttt tttattaacg
     4621 atacaaacca aacgcaacaa gaacatttgt agtattatct ataattgaaa acgcgtagtt
     4681 ataatcgctg aggtaatatt taaaatcatt ttcaaatgat tcacagttaa tttgcgacaa
     4741 tataatttta ttttcacata aactagacgc cttgtcgtct tcttcttcgt attccttctc
     4801 tttttcattt ttctcctcat aaaaattaac atagttatta tcgtatccat atatgtatct
     4861 atcgtataga gtaaattttt tgttgtcata aatatatatg tcttttttaa tggggtgtat
     4921 agtaccgctg cgcatagttt ttctgtaatt tacaacagtg ctattttctg gtagttcttc
     4981 ggagtgtgtt gctttaatta ttaaatttat ataatcaatg aatttgggat cgtcggtttt
     5041 gtacaatatg ttgccggcat agtacgcagc ttcttctagt tcaattacac cattttttag
     5101 cagcaccgga ttaacataac tttccaaaat gttgtacgaa ccgttaaaca aaaacagttc
     5161 acctcccttt tctatactat tgtctgcgag cagttgtttg ttgttaaaaa taacagccat
     5221 tgtaatgaga cgcacaaact aatatcacaa gctgccacca ttttgttcgc ccaggactct
     5281 agctatagtt ctagtggttg gctacgtata ctccggaata ttaatagatc atggagataa
     5341 ttaaaatgat aaccatctcg caaataaata agtattttac tgttttcgta acagttttgt
     5401 aataaaaaaa cctataaata ttccggatta ttcataccgt cccaccatcg ggcgcggatc
     5461 gccaccatgg gc
//
