LOCUS       sequence-232310-        6027 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 6027)
  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 6027)
  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..6027
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      42..51
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             54..707
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     665..687
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             714..737
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             846..1151
                     /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     1030..1049
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    1324..1458
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1361..1380)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1415..1434
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1531..1903
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(2255..2272)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2365..2953)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2445..2464)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3124..3915,3916..3984))
                     /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     3747..3766
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3985..4089)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4390..4412
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4404..4421
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4405..4421
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4432..4451
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4432..4450
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5061..5185
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5223..5828)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     promoter        5936..6027
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     5952..5969
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
ORIGIN
        1 attccggatt attcataccg tcccaccatc gggcgcggat cgccaccatg ggcatgtccc
       61 ctatactagg ttattggaaa attaagggcc ttgtgcaacc cactcgactt cttttggaat
      121 atcttgaaga aaaatatgaa gagcatttgt atgagcgcga tgaaggtgat aaatggcgaa
      181 acaaaaagtt tgaattgggt ttggagtttc ccaatcttcc ttattatatt gatggtgatg
      241 ttaaattaac acagtctatg gccatcatac gttatatagc tgacaagcac aacatgttgg
      301 gtggttgtcc aaaagagcgt gcagagattt caatgcttga aggagcggtt ttggatatta
      361 gatacggtgt ttcgagaatt gcatatagta aagactttga aactctcaaa gttgattttc
      421 ttagcaagct acctgaaatg ctgaaaatgt tcgaagatcg tttatgtcat aaaacatatt
      481 taaatggtga tcatgtaacc catcctgact tcatgttgta tgacgctctt gatgttgttt
      541 tatacatgga cccaatgtgc ctggatgcgt tcccaaaatt agtttgtttt aaaaaacgta
      601 ttgaagctat cccacaaatt gataagtact tgaaatccag caagtatata gcatggcctt
      661 tgcagggctg gcaagccacg tttggtggtg gcgaccatcc tccaaaatcg gatctggaag
      721 ttctgttcca ggggcccgcg gccgcaagcg ctatggccac aacctgttac attgcacaag
      781 ataaaaatat atcatcacga acagtaaaac tgtctgctta cataaacagt aatacaaggg
      841 gtgttatgca gtttaaggtt tacacctata aaagagagag ccgttatcgt ctgtttgtgg
      901 atgtacagag tgatattatt gacacgcccg ggcgacggat ggtgatcccc ctggccagtg
      961 cccgtctgct gtcagataaa gtctcccgtg aactttaccc ggtggtgcat atcggggatg
     1021 aaagctggcg catgatgacc accgatatgg ccagtgtgcc ggtctccgtt atcggggaag
     1081 aagtggctga tctcagccac cgcgaaaatg acatcaaaaa cgccattaac ctgatgttct
     1141 ggggaatata aatgtcaggc tcccttatac acagccagtg gcgcgccgta atagaagctt
     1201 gtcgagaagt actagaggat cataatcagc cataccacat ttgtagaggt tttacttgct
     1261 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     1321 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     1381 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     1441 tcttatcatg tctggatctg atcactgctt gagcctagga gatccgaacc agataagtga
     1501 aagcctgcat ccgcagatct taattaataa aacacgatac attgttatta gtacatttat
     1561 taagcgctag attctgtgcg ttgttgattt acagacaatt gttgtacgta ttttaataat
     1621 tcattaaatt tataatcttt agggtggtat gttagagcga aaatcaaatg attttcagcg
     1681 tctttatatc tgaatttaaa tattaaatcc tcaatagatt tgtaaaatag gtttcgatta
     1741 gtttcaaaca agggttgttt ttccgaaccg atggctggac tatctaatgg attttcgctc
     1801 aacgccacaa aacttgccaa atcttgtagc agcaatctag ctttgtcgat attcgtttgt
     1861 gttttgtttt gtaataaagg ttcgacgtcg ttcaaaatat tatgcgcttt tgtatttctt
     1921 tcatcactgt cgttagtgta caattgactc gacgtaaaca cgttaaataa cgcttggaca
     1981 tatttaacat cgggcgtgtt agctttatta ggccgattat cgtcgtcgtc ccaaccctcg
     2041 tcgttagaag ttgcttccga agacgatttt gccatagcca cacgacgcct attaattgtg
     2101 tcggctaaca cgtccgcgat caaatttgta gttgagcttt ttgggcctca tgggccttcc
     2161 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa catggtcata
     2221 gctgtttcct tgcgtattgg gcgctctccg cttcctcgct cactgactcg ctgcgctcgg
     2281 tcgttcgggt aaagcctggg gtgcctaatg agcaaaaggc cagcaaaagg ccaggaaccg
     2341 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     2401 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2461 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     2521 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     2581 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     2641 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     2701 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     2761 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     2821 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     2881 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     2941 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     3001 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     3061 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     3121 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     3181 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     3241 ccccagtgct gcaatgatac cgcgagaacc acgctcaccg gctccagatt tatcagcaat
     3301 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     3361 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     3421 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     3481 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     3541 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     3601 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     3661 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     3721 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     3781 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     3841 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     3901 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     3961 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     4021 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     4081 ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta
     4141 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     4201 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtggccg ctacagggcg
     4261 ctcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgtttcggtg cgggcctctt
     4321 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
     4381 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgagcgcgac gtaatacgac
     4441 tcactatagg gcgaattggc ggaaggccgt caaggccgtg acgttaaaac tattaagcca
     4501 tccaatcgac cgttagtcga atcaggaccg ctggtgcgag aagccgcgaa gtatggcgaa
     4561 tgcatcgtat aacgtgtgga gtccgctcat tagagcgtca tgtttagaca agaaagctac
     4621 atatttaatt gatcccgatg attttattga taaattgacc ctaactccat acacggtatt
     4681 ctacaatggc ggggttttgg tcaaaatttc cggactgcga ttgtacatgc tgttaacggc
     4741 tccgcccact attaatgaaa ttaaaaattc caattttaaa aaacgcagca agagaaacat
     4801 ttgtatgaaa gaatgcgtag aaggaaagaa aaatgtcgtg gacatgctga acaacaagat
     4861 taatatgcct ccgtgtataa aaaaaatatt gaacgatttg aaagaaaaca atgtaccgcg
     4921 cggcggtatg tacaggaaga ggtttatact aaactgttac attgcaaacg tggtttcgtg
     4981 tgccaagtgt gaaaaccgat gtttaatcaa ggctctgacg catttctaca accacgactc
     5041 caagtgtgtg ggtgaagtca tgcatctttt aatcaaatcc caagatgtgt ataaaccacc
     5101 aaactgccaa aaaatgaaaa ctgtggacaa gctctgtccg tttgctggca actgcaaggg
     5161 tctcaatcct atttgtaatt attgaataat aaaacaatta taaatgctaa atttgttttt
     5221 tattaacgat acaaaccaaa cgcaacaaga acatttgtag tattatctat aattgaaaac
     5281 gcgtagttat aatcgctgag gtaatattta aaatcatttt caaatgattc acagttaatt
     5341 tgcgacaata taattttatt ttcacataaa ctagacgcct tgtcgtcttc ttcttcgtat
     5401 tccttctctt tttcattttt ctcctcataa aaattaacat agttattatc gtatccatat
     5461 atgtatctat cgtatagagt aaattttttg ttgtcataaa tatatatgtc ttttttaatg
     5521 gggtgtatag taccgctgcg catagttttt ctgtaattta caacagtgct attttctggt
     5581 agttcttcgg agtgtgttgc tttaattatt aaatttatat aatcaatgaa tttgggatcg
     5641 tcggttttgt acaatatgtt gccggcatag tacgcagctt cttctagttc aattacacca
     5701 ttttttagca gcaccggatt aacataactt tccaaaatgt tgtacgaacc gttaaacaaa
     5761 aacagttcac ctcccttttc tatactattg tctgcgagca gttgtttgtt gttaaaaata
     5821 acagccattg taatgagacg cacaaactaa tatcacaagc tgccaccatt ttgttcgccc
     5881 aggactctag ctatagttct agtggttggc tacgtatact ccggaatatt aatagatcat
     5941 ggagataatt aaaatgataa ccatctcgca aataaataag tattttactg ttttcgtaac
     6001 agttttgtaa taaaaaaacc tataaat
//
