LOCUS       sequence-224697-        5467 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with N-terminal HIS10, cleavable with 3C 
            protease and N-terminal YBBR tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5467)
  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 5467)
  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: pOCC218
FEATURES             Location/Qualifiers
     source          1..5467
                     /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..259
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             272..295
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             455..760
                     /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     639..658
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    933..1067
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(970..989)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1024..1043
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1140..1512
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(1864..1881)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1974..2562)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2054..2073)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2733..3524,3525..3593))
                     /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     3356..3375
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3595..3699)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4000..4022
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4014..4031
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4015..4031
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4042..4061
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4042..4060
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4671..4795
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(4833..5438)
                     /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 ccatcaccat caccatcaca gcagcggcag actggaagtt ctgttccagg ggccggacag
      301 tctggagttc attgcgagca agctcgctgg tagcggatct ggttcagcgg ccgcaagcgc
      361 tatggccaca acctgttaca ttgcacaaga taaaaatata tcatcacgaa cagtaaaact
      421 gtctgcttac ataaacagta atacaagggg tgttatgcag tttaaggttt acacctataa
      481 aagagagagc cgttatcgtc tgtttgtgga tgtacagagt gatattattg acacgcccgg
      541 gcgacggatg gtgatccccc tggccagtgc ccgtctgctg tcagataaag tctcccgtga
      601 actttacccg gtggtgcata tcggggatga aagctggcgc atgatgacca ccgatatggc
      661 cagtgtgccg gtctccgtta tcggggaaga agtggctgat ctcagccacc gcgaaaatga
      721 catcaaaaac gccattaacc tgatgttctg gggaatataa atgtcaggct cccttataca
      781 cagccagtgg cgcgccgtaa tagaagcttg tcgagaagta ctagaggatc ataatcagcc
      841 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
      901 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
      961 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     1021 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatctga tcactgcttg
     1081 agcctaggag atccgaacca gataagtgaa agcctgcatc cgcagatctt aattaataaa
     1141 acacgataca ttgttattag tacatttatt aagcgctaga ttctgtgcgt tgttgattta
     1201 cagacaattg ttgtacgtat tttaataatt cattaaattt ataatcttta gggtggtatg
     1261 ttagagcgaa aatcaaatga ttttcagcgt ctttatatct gaatttaaat attaaatcct
     1321 caatagattt gtaaaatagg tttcgattag tttcaaacaa gggttgtttt tccgaaccga
     1381 tggctggact atctaatgga ttttcgctca acgccacaaa acttgccaaa tcttgtagca
     1441 gcaatctagc tttgtcgata ttcgtttgtg ttttgttttg taataaaggt tcgacgtcgt
     1501 tcaaaatatt atgcgctttt gtatttcttt catcactgtc gttagtgtac aattgactcg
     1561 acgtaaacac gttaaataac gcttggacat atttaacatc gggcgtgtta gctttattag
     1621 gccgattatc gtcgtcgtcc caaccctcgt cgttagaagt tgcttccgaa gacgattttg
     1681 ccatagccac acgacgccta ttaattgtgt cggctaacac gtccgcgatc aaatttgtag
     1741 ttgagctttt tgggcctcat gggccttccg ctcactgccc gctttccagt cgggaaacct
     1801 gtcgtgccag ctgcattaac atggtcatag ctgtttcctt gcgtattggg cgctctccgc
     1861 ttcctcgctc actgactcgc tgcgctcggt cgttcgggta aagcctgggg tgcctaatga
     1921 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     1981 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2041 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2101 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2161 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2221 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2281 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2341 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2401 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     2461 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2521 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2581 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     2641 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     2701 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     2761 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     2821 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     2881 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     2941 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     3001 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg
     3061 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3121 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     3181 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     3241 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     3301 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat
     3361 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     3421 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     3481 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     3541 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact cattactctt
     3601 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     3661 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     3721 acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     3781 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     3841 gagatagggt tgagtggccg ctacagggcg ctcccattcg ccattcaggc tgcgcaactg
     3901 ttgggaaggg cgtttcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
     3961 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
     4021 cgacggccag tgagcgcgac gtaatacgac tcactatagg gcgaattggc ggaaggccgt
     4081 caaggccgtg acgttaaaac tattaagcca tccaatcgac cgttagtcga atcaggaccg
     4141 ctggtgcgag aagccgcgaa gtatggcgaa tgcatcgtat aacgtgtgga gtccgctcat
     4201 tagagcgtca tgtttagaca agaaagctac atatttaatt gatcccgatg attttattga
     4261 taaattgacc ctaactccat acacggtatt ctacaatggc ggggttttgg tcaaaatttc
     4321 cggactgcga ttgtacatgc tgttaacggc tccgcccact attaatgaaa ttaaaaattc
     4381 caattttaaa aaacgcagca agagaaacat ttgtatgaaa gaatgcgtag aaggaaagaa
     4441 aaatgtcgtg gacatgctga acaacaagat taatatgcct ccgtgtataa aaaaaatatt
     4501 gaacgatttg aaagaaaaca atgtaccgcg cggcggtatg tacaggaaga ggtttatact
     4561 aaactgttac attgcaaacg tggtttcgtg tgccaagtgt gaaaaccgat gtttaatcaa
     4621 ggctctgacg catttctaca accacgactc caagtgtgtg ggtgaagtca tgcatctttt
     4681 aatcaaatcc caagatgtgt ataaaccacc aaactgccaa aaaatgaaaa ctgtggacaa
     4741 gctctgtccg tttgctggca actgcaaggg tctcaatcct atttgtaatt attgaataat
     4801 aaaacaatta taaatgctaa atttgttttt tattaacgat acaaaccaaa cgcaacaaga
     4861 acatttgtag tattatctat aattgaaaac gcgtagttat aatcgctgag gtaatattta
     4921 aaatcatttt caaatgattc acagttaatt tgcgacaata taattttatt ttcacataaa
     4981 ctagacgcct tgtcgtcttc ttcttcgtat tccttctctt tttcattttt ctcctcataa
     5041 aaattaacat agttattatc gtatccatat atgtatctat cgtatagagt aaattttttg
     5101 ttgtcataaa tatatatgtc ttttttaatg gggtgtatag taccgctgcg catagttttt
     5161 ctgtaattta caacagtgct attttctggt agttcttcgg agtgtgttgc tttaattatt
     5221 aaatttatat aatcaatgaa tttgggatcg tcggttttgt acaatatgtt gccggcatag
     5281 tacgcagctt cttctagttc aattacacca ttttttagca gcaccggatt aacataactt
     5341 tccaaaatgt tgtacgaacc gttaaacaaa aacagttcac ctcccttttc tatactattg
     5401 tctgcgagca gttgtttgtt gttaaaaata acagccattg taatgagacg cacaaactaa
     5461 tatcaca
//
