LOCUS       sequence-224689-        6028 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with N-terminal GST, cleavable with 3C .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6028)
  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 6028)
  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: pOCC151
FEATURES             Location/Qualifiers
     source          1..6028
                     /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             224..877
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     835..857
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             884..907
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1016..1321
                     /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     1200..1219
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    1494..1628
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1531..1550)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1585..1604
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1701..2073
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(2425..2442)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2535..3123)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2615..2634)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3294..4085,4086..4154))
                     /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     3917..3936
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4156..4260)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4561..4583
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4575..4592
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4576..4592
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4603..4622
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4603..4621
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5232..5356
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5394..5999)
                     /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 ggcatgtccc ctatactagg
      241 ttattggaaa attaagggcc ttgtgcaacc cactcgactt cttttggaat atcttgaaga
      301 aaaatatgaa gagcatttgt atgagcgcga tgaaggtgat aaatggcgaa acaaaaagtt
      361 tgaattgggt ttggagtttc ccaatcttcc ttattatatt gatggtgatg ttaaattaac
      421 acagtctatg gccatcatac gttatatagc tgacaagcac aacatgttgg gtggttgtcc
      481 aaaagagcgt gcagagattt caatgcttga aggagcggtt ttggatatta gatacggtgt
      541 ttcgagaatt gcatatagta aagactttga aactctcaaa gttgattttc ttagcaagct
      601 acctgaaatg ctgaaaatgt tcgaagatcg tttatgtcat aaaacatatt taaatggtga
      661 tcatgtaacc catcctgact tcatgttgta tgacgctctt gatgttgttt tatacatgga
      721 cccaatgtgc ctggatgcgt tcccaaaatt agtttgtttt aaaaaacgta ttgaagctat
      781 cccacaaatt gataagtact tgaaatccag caagtatata gcatggcctt tgcagggctg
      841 gcaagccacg tttggtggtg gcgaccatcc tccaaaatcg gatctggaag ttctgttcca
      901 ggggcccgcg gccgcaagcg ctatggccac aacctgttac attgcacaag ataaaaatat
      961 atcatcacga acagtaaaac tgtctgctta cataaacagt aatacaaggg gtgttatgca
     1021 gtttaaggtt tacacctata aaagagagag ccgttatcgt ctgtttgtgg atgtacagag
     1081 tgatattatt gacacgcccg ggcgacggat ggtgatcccc ctggccagtg cccgtctgct
     1141 gtcagataaa gtctcccgtg aactttaccc ggtggtgcat atcggggatg aaagctggcg
     1201 catgatgacc accgatatgg ccagtgtgcc ggtctccgtt atcggggaag aagtggctga
     1261 tctcagccac cgcgaaaatg acatcaaaaa cgccattaac ctgatgttct ggggaatata
     1321 aatgtcaggc tcccttatac acagccagtg gcgcgccgta atagaagctt gtcgagaagt
     1381 actagaggat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     1441 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     1501 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     1561 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     1621 tctggatctg atcactgctt gagcctagga gatccgaacc agataagtga aagcctgcat
     1681 ccgcagatct taattaataa aacacgatac attgttatta gtacatttat taagcgctag
     1741 attctgtgcg ttgttgattt acagacaatt gttgtacgta ttttaataat tcattaaatt
     1801 tataatcttt agggtggtat gttagagcga aaatcaaatg attttcagcg tctttatatc
     1861 tgaatttaaa tattaaatcc tcaatagatt tgtaaaatag gtttcgatta gtttcaaaca
     1921 agggttgttt ttccgaaccg atggctggac tatctaatgg attttcgctc aacgccacaa
     1981 aacttgccaa atcttgtagc agcaatctag ctttgtcgat attcgtttgt gttttgtttt
     2041 gtaataaagg ttcgacgtcg ttcaaaatat tatgcgcttt tgtatttctt tcatcactgt
     2101 cgttagtgta caattgactc gacgtaaaca cgttaaataa cgcttggaca tatttaacat
     2161 cgggcgtgtt agctttatta ggccgattat cgtcgtcgtc ccaaccctcg tcgttagaag
     2221 ttgcttccga agacgatttt gccatagcca cacgacgcct attaattgtg tcggctaaca
     2281 cgtccgcgat caaatttgta gttgagcttt ttgggcctca tgggccttcc gctcactgcc
     2341 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa catggtcata gctgtttcct
     2401 tgcgtattgg gcgctctccg cttcctcgct cactgactcg ctgcgctcgg tcgttcgggt
     2461 aaagcctggg gtgcctaatg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2521 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2581 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2641 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2701 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     2761 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2821 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     2881 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     2941 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     3001 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3061 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3121 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3181 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     3241 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     3301 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     3361 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     3421 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     3481 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     3541 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     3601 gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     3661 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     3721 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     3781 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     3841 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     3901 ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     3961 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     4021 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     4081 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     4141 tgttgaatac tcattactct tcctttttca atattattga agcatttatc agggttattg
     4201 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     4261 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg
     4321 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     4381 tataaatcaa aagaatagac cgagataggg ttgagtggcc gctacagggc gctcccattc
     4441 gccattcagg ctgcgcaact gttgggaagg gcgtttcggt gcgggcctct tcgctattac
     4501 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     4561 cccagtcacg acgttgtaaa acgacggcca gtgagcgcga cgtaatacga ctcactatag
     4621 ggcgaattgg cggaaggccg tcaaggccgt gacgttaaaa ctattaagcc atccaatcga
     4681 ccgttagtcg aatcaggacc gctggtgcga gaagccgcga agtatggcga atgcatcgta
     4741 taacgtgtgg agtccgctca ttagagcgtc atgtttagac aagaaagcta catatttaat
     4801 tgatcccgat gattttattg ataaattgac cctaactcca tacacggtat tctacaatgg
     4861 cggggttttg gtcaaaattt ccggactgcg attgtacatg ctgttaacgg ctccgcccac
     4921 tattaatgaa attaaaaatt ccaattttaa aaaacgcagc aagagaaaca tttgtatgaa
     4981 agaatgcgta gaaggaaaga aaaatgtcgt ggacatgctg aacaacaaga ttaatatgcc
     5041 tccgtgtata aaaaaaatat tgaacgattt gaaagaaaac aatgtaccgc gcggcggtat
     5101 gtacaggaag aggtttatac taaactgtta cattgcaaac gtggtttcgt gtgccaagtg
     5161 tgaaaaccga tgtttaatca aggctctgac gcatttctac aaccacgact ccaagtgtgt
     5221 gggtgaagtc atgcatcttt taatcaaatc ccaagatgtg tataaaccac caaactgcca
     5281 aaaaatgaaa actgtggaca agctctgtcc gtttgctggc aactgcaagg gtctcaatcc
     5341 tatttgtaat tattgaataa taaaacaatt ataaatgcta aatttgtttt ttattaacga
     5401 tacaaaccaa acgcaacaag aacatttgta gtattatcta taattgaaaa cgcgtagtta
     5461 taatcgctga ggtaatattt aaaatcattt tcaaatgatt cacagttaat ttgcgacaat
     5521 ataattttat tttcacataa actagacgcc ttgtcgtctt cttcttcgta ttccttctct
     5581 ttttcatttt tctcctcata aaaattaaca tagttattat cgtatccata tatgtatcta
     5641 tcgtatagag taaatttttt gttgtcataa atatatatgt cttttttaat ggggtgtata
     5701 gtaccgctgc gcatagtttt tctgtaattt acaacagtgc tattttctgg tagttcttcg
     5761 gagtgtgttg ctttaattat taaatttata taatcaatga atttgggatc gtcggttttg
     5821 tacaatatgt tgccggcata gtacgcagct tcttctagtt caattacacc attttttagc
     5881 agcaccggat taacataact ttccaaaatg ttgtacgaac cgttaaacaa aaacagttca
     5941 cctccctttt ctatactatt gtctgcgagc agttgtttgt tgttaaaaat aacagccatt
     6001 gtaatgagac gcacaaacta atatcaca
//
