LOCUS       sequence-232329-        5466 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 5466)
  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 5466)
  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..5466
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..92
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     17..34
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     regulatory      134..143
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             152..181
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             194..217
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             377..682
                     /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     561..580
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    855..989
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(892..911)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     946..965
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1062..1434
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(1786..1803)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1896..2484)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1976..1995)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2655..3446,3447..3515))
                     /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     3278..3297
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3516..3620)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3921..3943
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3935..3952
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3936..3952
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3963..3982
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3963..3981
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4592..4716
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(4754..5359)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
ORIGIN
        1 atcatggaga taattaaaat gataaccatc tcgcaaataa ataagtattt tactgttttc
       61 gtaacagttt tgtaataaaa aaacctataa atattccgga ttattcatac cgtcccacca
      121 tcgggcgcgg atcgccacca tgggcagcag ccatcaccat caccatcacc atcaccatca
      181 cagcagcggc agactggaag ttctgttcca ggggccggac agtctggagt tcattgcgag
      241 caagctcgct ggtagcggat ctggttcagc ggccgcaagc gctatggcca caacctgtta
      301 cattgcacaa gataaaaata tatcatcacg aacagtaaaa ctgtctgctt acataaacag
      361 taatacaagg ggtgttatgc agtttaaggt ttacacctat aaaagagaga gccgttatcg
      421 tctgtttgtg gatgtacaga gtgatattat tgacacgccc gggcgacgga tggtgatccc
      481 cctggccagt gcccgtctgc tgtcagataa agtctcccgt gaactttacc cggtggtgca
      541 tatcggggat gaaagctggc gcatgatgac caccgatatg gccagtgtgc cggtctccgt
      601 tatcggggaa gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa acgccattaa
      661 cctgatgttc tggggaatat aaatgtcagg ctcccttata cacagccagt ggcgcgccgt
      721 aatagaagct tgtcgagaag tactagagga tcataatcag ccataccaca tttgtagagg
      781 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
      841 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
      901 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
      961 tcatcaatgt atcttatcat gtctggatct gatcactgct tgagcctagg agatccgaac
     1021 cagataagtg aaagcctgca tccgcagatc ttaattaata aaacacgata cattgttatt
     1081 agtacattta ttaagcgcta gattctgtgc gttgttgatt tacagacaat tgttgtacgt
     1141 attttaataa ttcattaaat ttataatctt tagggtggta tgttagagcg aaaatcaaat
     1201 gattttcagc gtctttatat ctgaatttaa atattaaatc ctcaatagat ttgtaaaata
     1261 ggtttcgatt agtttcaaac aagggttgtt tttccgaacc gatggctgga ctatctaatg
     1321 gattttcgct caacgccaca aaacttgcca aatcttgtag cagcaatcta gctttgtcga
     1381 tattcgtttg tgttttgttt tgtaataaag gttcgacgtc gttcaaaata ttatgcgctt
     1441 ttgtatttct ttcatcactg tcgttagtgt acaattgact cgacgtaaac acgttaaata
     1501 acgcttggac atatttaaca tcgggcgtgt tagctttatt aggccgatta tcgtcgtcgt
     1561 cccaaccctc gtcgttagaa gttgcttccg aagacgattt tgccatagcc acacgacgcc
     1621 tattaattgt gtcggctaac acgtccgcga tcaaatttgt agttgagctt tttgggcctc
     1681 atgggccttc cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     1741 acatggtcat agctgtttcc ttgcgtattg ggcgctctcc gcttcctcgc tcactgactc
     1801 gctgcgctcg gtcgttcggg taaagcctgg ggtgcctaat gagcaaaagg ccagcaaaag
     1861 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     1921 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     1981 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     2041 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     2101 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     2161 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     2221 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     2281 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     2341 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     2401 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     2461 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     2521 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     2581 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     2641 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     2701 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     2761 ttaccatctg gccccagtgc tgcaatgata ccgcgagaac cacgctcacc ggctccagat
     2821 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     2881 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     2941 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     3001 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     3061 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     3121 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     3181 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     3241 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     3301 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     3361 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     3421 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     3481 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     3541 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     3601 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta
     3661 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     3721 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtggcc
     3781 gctacagggc gctcccattc gccattcagg ctgcgcaact gttgggaagg gcgtttcggt
     3841 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     3901 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgagcgcga
     3961 cgtaatacga ctcactatag ggcgaattgg cggaaggccg tcaaggccgt gacgttaaaa
     4021 ctattaagcc atccaatcga ccgttagtcg aatcaggacc gctggtgcga gaagccgcga
     4081 agtatggcga atgcatcgta taacgtgtgg agtccgctca ttagagcgtc atgtttagac
     4141 aagaaagcta catatttaat tgatcccgat gattttattg ataaattgac cctaactcca
     4201 tacacggtat tctacaatgg cggggttttg gtcaaaattt ccggactgcg attgtacatg
     4261 ctgttaacgg ctccgcccac tattaatgaa attaaaaatt ccaattttaa aaaacgcagc
     4321 aagagaaaca tttgtatgaa agaatgcgta gaaggaaaga aaaatgtcgt ggacatgctg
     4381 aacaacaaga ttaatatgcc tccgtgtata aaaaaaatat tgaacgattt gaaagaaaac
     4441 aatgtaccgc gcggcggtat gtacaggaag aggtttatac taaactgtta cattgcaaac
     4501 gtggtttcgt gtgccaagtg tgaaaaccga tgtttaatca aggctctgac gcatttctac
     4561 aaccacgact ccaagtgtgt gggtgaagtc atgcatcttt taatcaaatc ccaagatgtg
     4621 tataaaccac caaactgcca aaaaatgaaa actgtggaca agctctgtcc gtttgctggc
     4681 aactgcaagg gtctcaatcc tatttgtaat tattgaataa taaaacaatt ataaatgcta
     4741 aatttgtttt ttattaacga tacaaaccaa acgcaacaag aacatttgta gtattatcta
     4801 taattgaaaa cgcgtagtta taatcgctga ggtaatattt aaaatcattt tcaaatgatt
     4861 cacagttaat ttgcgacaat ataattttat tttcacataa actagacgcc ttgtcgtctt
     4921 cttcttcgta ttccttctct ttttcatttt tctcctcata aaaattaaca tagttattat
     4981 cgtatccata tatgtatcta tcgtatagag taaatttttt gttgtcataa atatatatgt
     5041 cttttttaat ggggtgtata gtaccgctgc gcatagtttt tctgtaattt acaacagtgc
     5101 tattttctgg tagttcttcg gagtgtgttg ctttaattat taaatttata taatcaatga
     5161 atttgggatc gtcggttttg tacaatatgt tgccggcata gtacgcagct tcttctagtt
     5221 caattacacc attttttagc agcaccggat taacataact ttccaaaatg ttgtacgaac
     5281 cgttaaacaa aaacagttca cctccctttt ctatactatt gtctgcgagc agttgtttgt
     5341 tgttaaaaat aacagccatt gtaatgagac gcacaaacta atatcacaag ctgccaccat
     5401 tttgttcgcc caggactcta gctatagttc tagtggttgg ctacgtatac tccggaatat
     5461 taatag
//
