LOCUS       sequence-232331-        5406 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with N-terminal sortase site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5406)
  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 5406)
  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: pOCC114
FEATURES             Location/Qualifiers
     source          1..5406
                     /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..110
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             225..530
                     /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     409..428
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    703..837
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(740..759)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     794..813
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           910..1282
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(1634..1651)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1744..2332)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1824..1843)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2503..3294,3295..3363))
                     /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     3126..3145
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3364..3468)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3769..3791
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3783..3800
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3784..3800
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3811..3830
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3811..3829
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4440..4564
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(4602..5207)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     promoter        5315..5406
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     5331..5348
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
ORIGIN
        1 attccggatt attcataccg tcccaccatc gggcgcggat cgccaccatg ggcagcagcc
       61 atcaccatca ccatcacagc agcggcagag aaaacttgta tttccagggc ggtggcgcgg
      121 ccgcaagcgc tatggccaca acctgttaca ttgcacaaga taaaaatata tcatcacgaa
      181 cagtaaaact gtctgcttac ataaacagta atacaagggg tgttatgcag tttaaggttt
      241 acacctataa aagagagagc cgttatcgtc tgtttgtgga tgtacagagt gatattattg
      301 acacgcccgg gcgacggatg gtgatccccc tggccagtgc ccgtctgctg tcagataaag
      361 tctcccgtga actttacccg gtggtgcata tcggggatga aagctggcgc atgatgacca
      421 ccgatatggc cagtgtgccg gtctccgtta tcggggaaga agtggctgat ctcagccacc
      481 gcgaaaatga catcaaaaac gccattaacc tgatgttctg gggaatataa atgtcaggct
      541 cccttataca cagccagtgg cgcgccgtaa tagaagcttg tcgagaagta ctagaggatc
      601 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
      661 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
      721 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
      781 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatctga
      841 tcactgcttg agcctaggag atccgaacca gataagtgaa agcctgcatc cgcagatctt
      901 aattaataaa acacgataca ttgttattag tacatttatt aagcgctaga ttctgtgcgt
      961 tgttgattta cagacaattg ttgtacgtat tttaataatt cattaaattt ataatcttta
     1021 gggtggtatg ttagagcgaa aatcaaatga ttttcagcgt ctttatatct gaatttaaat
     1081 attaaatcct caatagattt gtaaaatagg tttcgattag tttcaaacaa gggttgtttt
     1141 tccgaaccga tggctggact atctaatgga ttttcgctca acgccacaaa acttgccaaa
     1201 tcttgtagca gcaatctagc tttgtcgata ttcgtttgtg ttttgttttg taataaaggt
     1261 tcgacgtcgt tcaaaatatt atgcgctttt gtatttcttt catcactgtc gttagtgtac
     1321 aattgactcg acgtaaacac gttaaataac gcttggacat atttaacatc gggcgtgtta
     1381 gctttattag gccgattatc gtcgtcgtcc caaccctcgt cgttagaagt tgcttccgaa
     1441 gacgattttg ccatagccac acgacgccta ttaattgtgt cggctaacac gtccgcgatc
     1501 aaatttgtag ttgagctttt tgggcctcat gggccttccg ctcactgccc gctttccagt
     1561 cgggaaacct gtcgtgccag ctgcattaac atggtcatag ctgtttcctt gcgtattggg
     1621 cgctctccgc ttcctcgctc actgactcgc tgcgctcggt cgttcgggta aagcctgggg
     1681 tgcctaatga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1741 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1801 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1861 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1921 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     1981 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2041 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2101 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2161 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     2221 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2281 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2341 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     2401 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2461 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2521 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     2581 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     2641 gcgagaacca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     2701 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     2761 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     2821 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     2881 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     2941 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3001 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     3061 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     3121 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3181 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3241 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3301 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     3361 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     3421 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     3481 aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     3541 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     3601 gaatagaccg agatagggtt gagtggccgc tacagggcgc tcccattcgc cattcaggct
     3661 gcgcaactgt tgggaagggc gtttcggtgc gggcctcttc gctattacgc cagctggcga
     3721 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     3781 gttgtaaaac gacggccagt gagcgcgacg taatacgact cactataggg cgaattggcg
     3841 gaaggccgtc aaggccgtga cgttaaaact attaagccat ccaatcgacc gttagtcgaa
     3901 tcaggaccgc tggtgcgaga agccgcgaag tatggcgaat gcatcgtata acgtgtggag
     3961 tccgctcatt agagcgtcat gtttagacaa gaaagctaca tatttaattg atcccgatga
     4021 ttttattgat aaattgaccc taactccata cacggtattc tacaatggcg gggttttggt
     4081 caaaatttcc ggactgcgat tgtacatgct gttaacggct ccgcccacta ttaatgaaat
     4141 taaaaattcc aattttaaaa aacgcagcaa gagaaacatt tgtatgaaag aatgcgtaga
     4201 aggaaagaaa aatgtcgtgg acatgctgaa caacaagatt aatatgcctc cgtgtataaa
     4261 aaaaatattg aacgatttga aagaaaacaa tgtaccgcgc ggcggtatgt acaggaagag
     4321 gtttatacta aactgttaca ttgcaaacgt ggtttcgtgt gccaagtgtg aaaaccgatg
     4381 tttaatcaag gctctgacgc atttctacaa ccacgactcc aagtgtgtgg gtgaagtcat
     4441 gcatctttta atcaaatccc aagatgtgta taaaccacca aactgccaaa aaatgaaaac
     4501 tgtggacaag ctctgtccgt ttgctggcaa ctgcaagggt ctcaatccta tttgtaatta
     4561 ttgaataata aaacaattat aaatgctaaa tttgtttttt attaacgata caaaccaaac
     4621 gcaacaagaa catttgtagt attatctata attgaaaacg cgtagttata atcgctgagg
     4681 taatatttaa aatcattttc aaatgattca cagttaattt gcgacaatat aattttattt
     4741 tcacataaac tagacgcctt gtcgtcttct tcttcgtatt ccttctcttt ttcatttttc
     4801 tcctcataaa aattaacata gttattatcg tatccatata tgtatctatc gtatagagta
     4861 aattttttgt tgtcataaat atatatgtct tttttaatgg ggtgtatagt accgctgcgc
     4921 atagtttttc tgtaatttac aacagtgcta ttttctggta gttcttcgga gtgtgttgct
     4981 ttaattatta aatttatata atcaatgaat ttgggatcgt cggttttgta caatatgttg
     5041 ccggcatagt acgcagcttc ttctagttca attacaccat tttttagcag caccggatta
     5101 acataacttt ccaaaatgtt gtacgaaccg ttaaacaaaa acagttcacc tcccttttct
     5161 atactattgt ctgcgagcag ttgtttgttg ttaaaaataa cagccattgt aatgagacgc
     5221 acaaactaat atcacaagct gccaccattt tgttcgccca ggactctagc tatagttcta
     5281 gtggttggct acgtatactc cggaatatta atagatcatg gagataatta aaatgataac
     5341 catctcgcaa ataaataagt attttactgt tttcgtaaca gttttgtaat aaaaaaacct
     5401 ataaat
//
