LOCUS       sequence-224691-        5458 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with N-terminal TwinStrep, cleavable with 
            3C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5458)
  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 5458)
  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: pOCC153
FEATURES             Location/Qualifiers
     source          1..5458
                     /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             join(224..229,230..253,254..289,290..313)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="SAWSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     CDS             314..337
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             446..751
                     /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     630..649
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    924..1058
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(961..980)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1015..1034
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1131..1503
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(1855..1872)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1965..2553)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2045..2064)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2724..3515,3516..3584))
                     /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     3347..3366
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3586..3690)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3991..4013
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4005..4022
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4006..4022
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4033..4052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4033..4051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4662..4786
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(4824..5429)
                     /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 ggcagcgctt ggagccatcc
      241 gcagtttgaa aaaggtggtg gtagcggtgg tggttcaggt ggtagtgcat ggtcacaccc
      301 tcagtttgag aaactggaag ttctgtttca gggtcctgcg gccgcaagcg ctatggccac
      361 aacctgttac attgcacaag ataaaaatat atcatcacga acagtaaaac tgtctgctta
      421 cataaacagt aatacaaggg gtgttatgca gtttaaggtt tacacctata aaagagagag
      481 ccgttatcgt ctgtttgtgg atgtacagag tgatattatt gacacgcccg ggcgacggat
      541 ggtgatcccc ctggccagtg cccgtctgct gtcagataaa gtctcccgtg aactttaccc
      601 ggtggtgcat atcggggatg aaagctggcg catgatgacc accgatatgg ccagtgtgcc
      661 ggtctccgtt atcggggaag aagtggctga tctcagccac cgcgaaaatg acatcaaaaa
      721 cgccattaac ctgatgttct ggggaatata aatgtcaggc tcccttatac acagccagtg
      781 gcgcgccgta atagaagctt gtcgagaagt actagaggat cataatcagc cataccacat
      841 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
      901 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
      961 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     1021 tgtccaaact catcaatgta tcttatcatg tctggatctg atcactgctt gagcctagga
     1081 gatccgaacc agataagtga aagcctgcat ccgcagatct taattaataa aacacgatac
     1141 attgttatta gtacatttat taagcgctag attctgtgcg ttgttgattt acagacaatt
     1201 gttgtacgta ttttaataat tcattaaatt tataatcttt agggtggtat gttagagcga
     1261 aaatcaaatg attttcagcg tctttatatc tgaatttaaa tattaaatcc tcaatagatt
     1321 tgtaaaatag gtttcgatta gtttcaaaca agggttgttt ttccgaaccg atggctggac
     1381 tatctaatgg attttcgctc aacgccacaa aacttgccaa atcttgtagc agcaatctag
     1441 ctttgtcgat attcgtttgt gttttgtttt gtaataaagg ttcgacgtcg ttcaaaatat
     1501 tatgcgcttt tgtatttctt tcatcactgt cgttagtgta caattgactc gacgtaaaca
     1561 cgttaaataa cgcttggaca tatttaacat cgggcgtgtt agctttatta ggccgattat
     1621 cgtcgtcgtc ccaaccctcg tcgttagaag ttgcttccga agacgatttt gccatagcca
     1681 cacgacgcct attaattgtg tcggctaaca cgtccgcgat caaatttgta gttgagcttt
     1741 ttgggcctca tgggccttcc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     1801 gctgcattaa catggtcata gctgtttcct tgcgtattgg gcgctctccg cttcctcgct
     1861 cactgactcg ctgcgctcgg tcgttcgggt aaagcctggg gtgcctaatg agcaaaaggc
     1921 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     1981 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2041 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2101 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2161 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2221 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2281 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2341 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2401 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2461 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2521 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2581 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2641 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     2701 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     2761 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     2821 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     2881 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     2941 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3001 tcgccagtta atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc
     3061 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3121 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     3181 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     3241 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     3301 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca
     3361 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     3421 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     3481 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     3541 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcattactct tcctttttca
     3601 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     3661 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt
     3721 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     3781 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     3841 ttgagtggcc gctacagggc gctcccattc gccattcagg ctgcgcaact gttgggaagg
     3901 gcgtttcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     3961 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     4021 gtgagcgcga cgtaatacga ctcactatag ggcgaattgg cggaaggccg tcaaggccgt
     4081 gacgttaaaa ctattaagcc atccaatcga ccgttagtcg aatcaggacc gctggtgcga
     4141 gaagccgcga agtatggcga atgcatcgta taacgtgtgg agtccgctca ttagagcgtc
     4201 atgtttagac aagaaagcta catatttaat tgatcccgat gattttattg ataaattgac
     4261 cctaactcca tacacggtat tctacaatgg cggggttttg gtcaaaattt ccggactgcg
     4321 attgtacatg ctgttaacgg ctccgcccac tattaatgaa attaaaaatt ccaattttaa
     4381 aaaacgcagc aagagaaaca tttgtatgaa agaatgcgta gaaggaaaga aaaatgtcgt
     4441 ggacatgctg aacaacaaga ttaatatgcc tccgtgtata aaaaaaatat tgaacgattt
     4501 gaaagaaaac aatgtaccgc gcggcggtat gtacaggaag aggtttatac taaactgtta
     4561 cattgcaaac gtggtttcgt gtgccaagtg tgaaaaccga tgtttaatca aggctctgac
     4621 gcatttctac aaccacgact ccaagtgtgt gggtgaagtc atgcatcttt taatcaaatc
     4681 ccaagatgtg tataaaccac caaactgcca aaaaatgaaa actgtggaca agctctgtcc
     4741 gtttgctggc aactgcaagg gtctcaatcc tatttgtaat tattgaataa taaaacaatt
     4801 ataaatgcta aatttgtttt ttattaacga tacaaaccaa acgcaacaag aacatttgta
     4861 gtattatcta taattgaaaa cgcgtagtta taatcgctga ggtaatattt aaaatcattt
     4921 tcaaatgatt cacagttaat ttgcgacaat ataattttat tttcacataa actagacgcc
     4981 ttgtcgtctt cttcttcgta ttccttctct ttttcatttt tctcctcata aaaattaaca
     5041 tagttattat cgtatccata tatgtatcta tcgtatagag taaatttttt gttgtcataa
     5101 atatatatgt cttttttaat ggggtgtata gtaccgctgc gcatagtttt tctgtaattt
     5161 acaacagtgc tattttctgg tagttcttcg gagtgtgttg ctttaattat taaatttata
     5221 taatcaatga atttgggatc gtcggttttg tacaatatgt tgccggcata gtacgcagct
     5281 tcttctagtt caattacacc attttttagc agcaccggat taacataact ttccaaaatg
     5341 ttgtacgaac cgttaaacaa aaacagttca cctccctttt ctatactatt gtctgcgagc
     5401 agttgtttgt tgttaaaaat aacagccatt gtaatgagac gcacaaacta atatcaca
//
