LOCUS       sequence-224666-        5398 bp    DNA     circular SYN 14-APR-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with C-terminal HIS6 tag, cleavable with 3C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5398)
  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 5398)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pOCC7
FEATURES             Location/Qualifiers
     source          1..5398
                     /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"
     regulatory      216..225
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             329..634
                     /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     513..532
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             672..695
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             705..722
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    864..998
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(901..920)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     955..974
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1071..1443
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(1795..1812)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1905..2493)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1985..2004)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2664..3455,3456..3524))
                     /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     3287..3306
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3526..3630)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3931..3953
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3945..3962
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3946..3962
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3973..3992
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3973..3991
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4602..4726
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(4764..5369)
                     /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 gcggccgcaa gcgctatggc
      241 cacaacctgt tacattgcac aagataaaaa tatatcatca cgaacagtaa aactgtctgc
      301 ttacataaac agtaatacaa ggggtgttat gcagtttaag gtttacacct ataaaagaga
      361 gagccgttat cgtctgtttg tggatgtaca gagtgatatt attgacacgc ccgggcgacg
      421 gatggtgatc cccctggcca gtgcccgtct gctgtcagat aaagtctccc gtgaacttta
      481 cccggtggtg catatcgggg atgaaagctg gcgcatgatg accaccgata tggccagtgt
      541 gccggtctcc gttatcgggg aagaagtggc tgatctcagc caccgcgaaa atgacatcaa
      601 aaacgccatt aacctgatgt tctggggaat ataaatgtca ggctccctta tacacagcca
      661 gtggcgcgcc gctggaagtt ctgttccagg ggccgggcag cagccatcac catcaccatc
      721 acagtggata atagaagctt gtcgagaagt actagaggat cataatcagc cataccacat
      781 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
      841 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
      901 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
      961 tgtccaaact catcaatgta tcttatcatg tctggatctg atcactgctt gagcctagga
     1021 gatccgaacc agataagtga aagcctgcat ccgcagatct taattaataa aacacgatac
     1081 attgttatta gtacatttat taagcgctag attctgtgcg ttgttgattt acagacaatt
     1141 gttgtacgta ttttaataat tcattaaatt tataatcttt agggtggtat gttagagcga
     1201 aaatcaaatg attttcagcg tctttatatc tgaatttaaa tattaaatcc tcaatagatt
     1261 tgtaaaatag gtttcgatta gtttcaaaca agggttgttt ttccgaaccg atggctggac
     1321 tatctaatgg attttcgctc aacgccacaa aacttgccaa atcttgtagc agcaatctag
     1381 ctttgtcgat attcgtttgt gttttgtttt gtaataaagg ttcgacgtcg ttcaaaatat
     1441 tatgcgcttt tgtatttctt tcatcactgt cgttagtgta caattgactc gacgtaaaca
     1501 cgttaaataa cgcttggaca tatttaacat cgggcgtgtt agctttatta ggccgattat
     1561 cgtcgtcgtc ccaaccctcg tcgttagaag ttgcttccga agacgatttt gccatagcca
     1621 cacgacgcct attaattgtg tcggctaaca cgtccgcgat caaatttgta gttgagcttt
     1681 ttgggcctca tgggccttcc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     1741 gctgcattaa catggtcata gctgtttcct tgcgtattgg gcgctctccg cttcctcgct
     1801 cactgactcg ctgcgctcgg tcgttcgggt aaagcctggg gtgcctaatg agcaaaaggc
     1861 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     1921 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     1981 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2041 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2101 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2161 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2221 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2281 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2341 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2401 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2461 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2521 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2581 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     2641 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     2701 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     2761 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     2821 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     2881 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     2941 tcgccagtta atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc
     3001 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3061 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     3121 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     3181 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     3241 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca
     3301 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     3361 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     3421 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     3481 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcattactct tcctttttca
     3541 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     3601 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt
     3661 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     3721 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     3781 ttgagtggcc gctacagggc gctcccattc gccattcagg ctgcgcaact gttgggaagg
     3841 gcgtttcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     3901 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     3961 gtgagcgcga cgtaatacga ctcactatag ggcgaattgg cggaaggccg tcaaggccgt
     4021 gacgttaaaa ctattaagcc atccaatcga ccgttagtcg aatcaggacc gctggtgcga
     4081 gaagccgcga agtatggcga atgcatcgta taacgtgtgg agtccgctca ttagagcgtc
     4141 atgtttagac aagaaagcta catatttaat tgatcccgat gattttattg ataaattgac
     4201 cctaactcca tacacggtat tctacaatgg cggggttttg gtcaaaattt ccggactgcg
     4261 attgtacatg ctgttaacgg ctccgcccac tattaatgaa attaaaaatt ccaattttaa
     4321 aaaacgcagc aagagaaaca tttgtatgaa agaatgcgta gaaggaaaga aaaatgtcgt
     4381 ggacatgctg aacaacaaga ttaatatgcc tccgtgtata aaaaaaatat tgaacgattt
     4441 gaaagaaaac aatgtaccgc gcggcggtat gtacaggaag aggtttatac taaactgtta
     4501 cattgcaaac gtggtttcgt gtgccaagtg tgaaaaccga tgtttaatca aggctctgac
     4561 gcatttctac aaccacgact ccaagtgtgt gggtgaagtc atgcatcttt taatcaaatc
     4621 ccaagatgtg tataaaccac caaactgcca aaaaatgaaa actgtggaca agctctgtcc
     4681 gtttgctggc aactgcaagg gtctcaatcc tatttgtaat tattgaataa taaaacaatt
     4741 ataaatgcta aatttgtttt ttattaacga tacaaaccaa acgcaacaag aacatttgta
     4801 gtattatcta taattgaaaa cgcgtagtta taatcgctga ggtaatattt aaaatcattt
     4861 tcaaatgatt cacagttaat ttgcgacaat ataattttat tttcacataa actagacgcc
     4921 ttgtcgtctt cttcttcgta ttccttctct ttttcatttt tctcctcata aaaattaaca
     4981 tagttattat cgtatccata tatgtatcta tcgtatagag taaatttttt gttgtcataa
     5041 atatatatgt cttttttaat ggggtgtata gtaccgctgc gcatagtttt tctgtaattt
     5101 acaacagtgc tattttctgg tagttcttcg gagtgtgttg ctttaattat taaatttata
     5161 taatcaatga atttgggatc gtcggttttg tacaatatgt tgccggcata gtacgcagct
     5221 tcttctagtt caattacacc attttttagc agcaccggat taacataact ttccaaaatg
     5281 ttgtacgaac cgttaaacaa aaacagttca cctccctttt ctatactatt gtctgcgagc
     5341 agttgtttgt tgttaaaaat aacagccatt gtaatgagac gcacaaacta atatcaca
//
