LOCUS       sequence-224662-        6151 bp    DNA     circular SYN 15-APR-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with N-terminal HIS6 tag, cleavable with 3C,
            and C-terminal monomeric GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6151)
  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 6151)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pOCC139
FEATURES             Location/Qualifiers
     source          1..6151
                     /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..283
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             392..697
                     /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     576..595
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             join(765..767,768..770,771..1484)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(810..831)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1071..1090)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1418..1439
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1617..1751
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1654..1673)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1708..1727
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1824..2196
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(2548..2565)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2658..3246)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2738..2757)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3417..4208,4209..4277))
                     /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     4040..4059
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4279..4383)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4684..4706
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4698..4715
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4699..4715
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4726..4745
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4726..4744
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5355..5479
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5517..6122)
                     /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 agcggcagac tggaagttct gttccagggg ccggcggccg caagcgctat
      301 ggccacaacc tgttacattg cacaagataa aaatatatca tcacgaacag taaaactgtc
      361 tgcttacata aacagtaata caaggggtgt tatgcagttt aaggtttaca cctataaaag
      421 agagagccgt tatcgtctgt ttgtggatgt acagagtgat attattgaca cgcccgggcg
      481 acggatggtg atccccctgg ccagtgcccg tctgctgtca gataaagtct cccgtgaact
      541 ttacccggtg gtgcatatcg gggatgaaag ctggcgcatg atgaccaccg atatggccag
      601 tgtgccggtc tccgttatcg gggaagaagt ggctgatctc agccaccgcg aaaatgacat
      661 caaaaacgcc attaacctga tgttctgggg aatataaatg tcaggctccc ttatacacag
      721 ccagtggcgc gccgggatcc gctggctccg ctgctggttc tggcatggtg agcaagggcg
      781 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
      841 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
      901 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
      961 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     1021 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     1081 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     1141 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     1201 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     1261 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     1321 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     1381 ccaaactgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     1441 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaatagaag cttgtcgaga
     1501 agtactagag gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     1561 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     1621 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     1681 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     1741 atgtctggat ctgatcactg cttgagccta ggagatccga accagataag tgaaagcctg
     1801 catccgcaga tcttaattaa taaaacacga tacattgtta ttagtacatt tattaagcgc
     1861 tagattctgt gcgttgttga tttacagaca attgttgtac gtattttaat aattcattaa
     1921 atttataatc tttagggtgg tatgttagag cgaaaatcaa atgattttca gcgtctttat
     1981 atctgaattt aaatattaaa tcctcaatag atttgtaaaa taggtttcga ttagtttcaa
     2041 acaagggttg tttttccgaa ccgatggctg gactatctaa tggattttcg ctcaacgcca
     2101 caaaacttgc caaatcttgt agcagcaatc tagctttgtc gatattcgtt tgtgttttgt
     2161 tttgtaataa aggttcgacg tcgttcaaaa tattatgcgc ttttgtattt ctttcatcac
     2221 tgtcgttagt gtacaattga ctcgacgtaa acacgttaaa taacgcttgg acatatttaa
     2281 catcgggcgt gttagcttta ttaggccgat tatcgtcgtc gtcccaaccc tcgtcgttag
     2341 aagttgcttc cgaagacgat tttgccatag ccacacgacg cctattaatt gtgtcggcta
     2401 acacgtccgc gatcaaattt gtagttgagc tttttgggcc tcatgggcct tccgctcact
     2461 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taacatggtc atagctgttt
     2521 ccttgcgtat tgggcgctct ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2581 ggtaaagcct ggggtgccta atgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     2641 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2701 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2761 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2821 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2881 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2941 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3001 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3061 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     3121 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3181 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     3241 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     3301 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     3361 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     3421 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     3481 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     3541 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     3601 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     3661 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     3721 gttgccattg ctgcaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     3781 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     3841 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     3901 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     3961 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     4021 tgcccggcgt caacacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     4081 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     4141 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     4201 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     4261 aaatgttgaa tactcattac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4321 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4381 gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc
     4441 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     4501 ccttataaat caaaagaata gaccgagata gggttgagtg gccgctacag ggcgctccca
     4561 ttcgccattc aggctgcgca actgttggga agggcgtttc ggtgcgggcc tcttcgctat
     4621 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     4681 tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgacgtaata cgactcacta
     4741 tagggcgaat tggcggaagg ccgtcaaggc cgtgacgtta aaactattaa gccatccaat
     4801 cgaccgttag tcgaatcagg accgctggtg cgagaagccg cgaagtatgg cgaatgcatc
     4861 gtataacgtg tggagtccgc tcattagagc gtcatgttta gacaagaaag ctacatattt
     4921 aattgatccc gatgatttta ttgataaatt gaccctaact ccatacacgg tattctacaa
     4981 tggcggggtt ttggtcaaaa tttccggact gcgattgtac atgctgttaa cggctccgcc
     5041 cactattaat gaaattaaaa attccaattt taaaaaacgc agcaagagaa acatttgtat
     5101 gaaagaatgc gtagaaggaa agaaaaatgt cgtggacatg ctgaacaaca agattaatat
     5161 gcctccgtgt ataaaaaaaa tattgaacga tttgaaagaa aacaatgtac cgcgcggcgg
     5221 tatgtacagg aagaggttta tactaaactg ttacattgca aacgtggttt cgtgtgccaa
     5281 gtgtgaaaac cgatgtttaa tcaaggctct gacgcatttc tacaaccacg actccaagtg
     5341 tgtgggtgaa gtcatgcatc ttttaatcaa atcccaagat gtgtataaac caccaaactg
     5401 ccaaaaaatg aaaactgtgg acaagctctg tccgtttgct ggcaactgca agggtctcaa
     5461 tcctatttgt aattattgaa taataaaaca attataaatg ctaaatttgt tttttattaa
     5521 cgatacaaac caaacgcaac aagaacattt gtagtattat ctataattga aaacgcgtag
     5581 ttataatcgc tgaggtaata tttaaaatca ttttcaaatg attcacagtt aatttgcgac
     5641 aatataattt tattttcaca taaactagac gccttgtcgt cttcttcttc gtattccttc
     5701 tctttttcat ttttctcctc ataaaaatta acatagttat tatcgtatcc atatatgtat
     5761 ctatcgtata gagtaaattt tttgttgtca taaatatata tgtctttttt aatggggtgt
     5821 atagtaccgc tgcgcatagt ttttctgtaa tttacaacag tgctattttc tggtagttct
     5881 tcggagtgtg ttgctttaat tattaaattt atataatcaa tgaatttggg atcgtcggtt
     5941 ttgtacaata tgttgccggc atagtacgca gcttcttcta gttcaattac accatttttt
     6001 agcagcaccg gattaacata actttccaaa atgttgtacg aaccgttaaa caaaaacagt
     6061 tcacctccct tttctatact attgtctgcg agcagttgtt tgttgttaaa aataacagcc
     6121 attgtaatga gacgcacaaa ctaatatcac a
//
