LOCUS       sequence-224694-        5407 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 5407)
  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 5407)
  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..5407
                     /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..280
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             395..700
                     /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     579..598
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    873..1007
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(910..929)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     964..983
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1080..1452
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(1804..1821)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1914..2502)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1994..2013)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2673..3464,3465..3533))
                     /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     3296..3315
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3535..3639)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3940..3962
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3954..3971
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3955..3971
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3982..4001
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3982..4000
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4611..4735
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(4773..5378)
                     /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 agcggcagag aaaacttgta tttccagggc ggtggcgcgg ccgcaagcgc
      301 tatggccaca acctgttaca ttgcacaaga taaaaatata tcatcacgaa cagtaaaact
      361 gtctgcttac ataaacagta atacaagggg tgttatgcag tttaaggttt acacctataa
      421 aagagagagc cgttatcgtc tgtttgtgga tgtacagagt gatattattg acacgcccgg
      481 gcgacggatg gtgatccccc tggccagtgc ccgtctgctg tcagataaag tctcccgtga
      541 actttacccg gtggtgcata tcggggatga aagctggcgc atgatgacca ccgatatggc
      601 cagtgtgccg gtctccgtta tcggggaaga agtggctgat ctcagccacc gcgaaaatga
      661 catcaaaaac gccattaacc tgatgttctg gggaatataa atgtcaggct cccttataca
      721 cagccagtgg cgcgccgtaa tagaagcttg tcgagaagta ctagaggatc ataatcagcc
      781 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
      841 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
      901 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
      961 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatctga tcactgcttg
     1021 agcctaggag atccgaacca gataagtgaa agcctgcatc cgcagatctt aattaataaa
     1081 acacgataca ttgttattag tacatttatt aagcgctaga ttctgtgcgt tgttgattta
     1141 cagacaattg ttgtacgtat tttaataatt cattaaattt ataatcttta gggtggtatg
     1201 ttagagcgaa aatcaaatga ttttcagcgt ctttatatct gaatttaaat attaaatcct
     1261 caatagattt gtaaaatagg tttcgattag tttcaaacaa gggttgtttt tccgaaccga
     1321 tggctggact atctaatgga ttttcgctca acgccacaaa acttgccaaa tcttgtagca
     1381 gcaatctagc tttgtcgata ttcgtttgtg ttttgttttg taataaaggt tcgacgtcgt
     1441 tcaaaatatt atgcgctttt gtatttcttt catcactgtc gttagtgtac aattgactcg
     1501 acgtaaacac gttaaataac gcttggacat atttaacatc gggcgtgtta gctttattag
     1561 gccgattatc gtcgtcgtcc caaccctcgt cgttagaagt tgcttccgaa gacgattttg
     1621 ccatagccac acgacgccta ttaattgtgt cggctaacac gtccgcgatc aaatttgtag
     1681 ttgagctttt tgggcctcat gggccttccg ctcactgccc gctttccagt cgggaaacct
     1741 gtcgtgccag ctgcattaac atggtcatag ctgtttcctt gcgtattggg cgctctccgc
     1801 ttcctcgctc actgactcgc tgcgctcggt cgttcgggta aagcctgggg tgcctaatga
     1861 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     1921 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     1981 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2041 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2101 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2161 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2221 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2281 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2341 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     2401 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2461 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2521 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     2581 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     2641 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     2701 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     2761 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     2821 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     2881 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     2941 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg
     3001 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3061 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     3121 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     3181 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     3241 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat
     3301 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     3361 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     3421 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     3481 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact cattactctt
     3541 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     3601 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     3661 acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     3721 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     3781 gagatagggt tgagtggccg ctacagggcg ctcccattcg ccattcaggc tgcgcaactg
     3841 ttgggaaggg cgtttcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
     3901 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
     3961 cgacggccag tgagcgcgac gtaatacgac tcactatagg gcgaattggc ggaaggccgt
     4021 caaggccgtg acgttaaaac tattaagcca tccaatcgac cgttagtcga atcaggaccg
     4081 ctggtgcgag aagccgcgaa gtatggcgaa tgcatcgtat aacgtgtgga gtccgctcat
     4141 tagagcgtca tgtttagaca agaaagctac atatttaatt gatcccgatg attttattga
     4201 taaattgacc ctaactccat acacggtatt ctacaatggc ggggttttgg tcaaaatttc
     4261 cggactgcga ttgtacatgc tgttaacggc tccgcccact attaatgaaa ttaaaaattc
     4321 caattttaaa aaacgcagca agagaaacat ttgtatgaaa gaatgcgtag aaggaaagaa
     4381 aaatgtcgtg gacatgctga acaacaagat taatatgcct ccgtgtataa aaaaaatatt
     4441 gaacgatttg aaagaaaaca atgtaccgcg cggcggtatg tacaggaaga ggtttatact
     4501 aaactgttac attgcaaacg tggtttcgtg tgccaagtgt gaaaaccgat gtttaatcaa
     4561 ggctctgacg catttctaca accacgactc caagtgtgtg ggtgaagtca tgcatctttt
     4621 aatcaaatcc caagatgtgt ataaaccacc aaactgccaa aaaatgaaaa ctgtggacaa
     4681 gctctgtccg tttgctggca actgcaaggg tctcaatcct atttgtaatt attgaataat
     4741 aaaacaatta taaatgctaa atttgttttt tattaacgat acaaaccaaa cgcaacaaga
     4801 acatttgtag tattatctat aattgaaaac gcgtagttat aatcgctgag gtaatattta
     4861 aaatcatttt caaatgattc acagttaatt tgcgacaata taattttatt ttcacataaa
     4921 ctagacgcct tgtcgtcttc ttcttcgtat tccttctctt tttcattttt ctcctcataa
     4981 aaattaacat agttattatc gtatccatat atgtatctat cgtatagagt aaattttttg
     5041 ttgtcataaa tatatatgtc ttttttaatg gggtgtatag taccgctgcg catagttttt
     5101 ctgtaattta caacagtgct attttctggt agttcttcgg agtgtgttgc tttaattatt
     5161 aaatttatat aatcaatgaa tttgggatcg tcggttttgt acaatatgtt gccggcatag
     5221 tacgcagctt cttctagttc aattacacca ttttttagca gcaccggatt aacataactt
     5281 tccaaaatgt tgtacgaacc gttaaacaaa aacagttcac ctcccttttc tatactattg
     5341 tctgcgagca gttgtttgtt gttaaaaata acagccattg taatgagacg cacaaactaa
     5401 tatcaca
//
