LOCUS       sequence-232307-        6063 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with N-terminal HIS6, cleavable with 3C , 
            and N-terminal GST.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6063)
  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 6063)
  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: pOCC81
FEATURES             Location/Qualifiers
     source          1..6063
                     /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..169
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             182..205
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             206..859
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     817..839
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             974..1279
                     /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     1158..1177
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    1452..1586
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1489..1508)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1543..1562
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1659..2031
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(2383..2400)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2493..3081)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2573..2592)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3252..4043,4044..4112))
                     /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     3875..3894
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4113..4217)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4518..4540
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4532..4549
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4533..4549
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4560..4579
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4560..4578
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5189..5313
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5351..5956)
                     /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 caccatcaca gcagcggcag
      181 actggaagtt ctgttccagg ggccgatgtc ccctatacta ggttattgga aaattaaggg
      241 ccttgtgcaa cccactcgac ttcttttgga atatcttgaa gaaaaatatg aagagcattt
      301 gtatgagcgc gatgaaggtg ataaatggcg aaacaaaaag tttgaattgg gtttggagtt
      361 tcccaatctt ccttattata ttgatggtga tgttaaatta acacagtcta tggccatcat
      421 acgttatata gctgacaagc acaacatgtt gggtggttgt ccaaaagagc gtgcagagat
      481 ttcaatgctt gaaggagcgg ttttggatat tagatacggt gtttcgagaa ttgcatatag
      541 taaagacttt gaaactctca aagttgattt tcttagcaag ctacctgaaa tgctgaaaat
      601 gttcgaagat cgtttatgtc ataaaacata tttaaatggt gatcatgtaa cccatcctga
      661 cttcatgttg tatgacgctc ttgatgttgt tttatacatg gacccaatgt gcctggatgc
      721 gttcccaaaa ttagtttgtt ttaaaaaacg tattgaagct atcccacaaa ttgataagta
      781 cttgaaatcc agcaagtata tagcatggcc tttgcagggc tggcaagcca cgtttggtgg
      841 tggcgaccat cctccaaaat cggatgcggc cgcaagcgct atggccacaa cctgttacat
      901 tgcacaagat aaaaatatat catcacgaac agtaaaactg tctgcttaca taaacagtaa
      961 tacaaggggt gttatgcagt ttaaggttta cacctataaa agagagagcc gttatcgtct
     1021 gtttgtggat gtacagagtg atattattga cacgcccggg cgacggatgg tgatccccct
     1081 ggccagtgcc cgtctgctgt cagataaagt ctcccgtgaa ctttacccgg tggtgcatat
     1141 cggggatgaa agctggcgca tgatgaccac cgatatggcc agtgtgccgg tctccgttat
     1201 cggggaagaa gtggctgatc tcagccaccg cgaaaatgac atcaaaaacg ccattaacct
     1261 gatgttctgg ggaatataaa tgtcaggctc ccttatacac agccagtggc gcgccgtaat
     1321 agaagcttgt cgagaagtac tagaggatca taatcagcca taccacattt gtagaggttt
     1381 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     1441 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     1501 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     1561 tcaatgtatc ttatcatgtc tggatctgat cactgcttga gcctaggaga tccgaaccag
     1621 ataagtgaaa gcctgcatcc gcagatctta attaataaaa cacgatacat tgttattagt
     1681 acatttatta agcgctagat tctgtgcgtt gttgatttac agacaattgt tgtacgtatt
     1741 ttaataattc attaaattta taatctttag ggtggtatgt tagagcgaaa atcaaatgat
     1801 tttcagcgtc tttatatctg aatttaaata ttaaatcctc aatagatttg taaaataggt
     1861 ttcgattagt ttcaaacaag ggttgttttt ccgaaccgat ggctggacta tctaatggat
     1921 tttcgctcaa cgccacaaaa cttgccaaat cttgtagcag caatctagct ttgtcgatat
     1981 tcgtttgtgt tttgttttgt aataaaggtt cgacgtcgtt caaaatatta tgcgcttttg
     2041 tatttctttc atcactgtcg ttagtgtaca attgactcga cgtaaacacg ttaaataacg
     2101 cttggacata tttaacatcg ggcgtgttag ctttattagg ccgattatcg tcgtcgtccc
     2161 aaccctcgtc gttagaagtt gcttccgaag acgattttgc catagccaca cgacgcctat
     2221 taattgtgtc ggctaacacg tccgcgatca aatttgtagt tgagcttttt gggcctcatg
     2281 ggccttccgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaaca
     2341 tggtcatagc tgtttccttg cgtattgggc gctctccgct tcctcgctca ctgactcgct
     2401 gcgctcggtc gttcgggtaa agcctggggt gcctaatgag caaaaggcca gcaaaaggcc
     2461 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2521 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2581 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2641 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2701 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2761 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     2821 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     2881 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     2941 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3001 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3061 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3121 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3181 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3241 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3301 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     3361 ccatctggcc ccagtgctgc aatgataccg cgagaaccac gctcaccggc tccagattta
     3421 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3481 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     3541 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     3601 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     3661 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     3721 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     3781 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     3841 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     3901 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     3961 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4021 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4081 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     4141 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     4201 caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata
     4261 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     4321 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtggccgct
     4381 acagggcgct cccattcgcc attcaggctg cgcaactgtt gggaagggcg tttcggtgcg
     4441 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     4501 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg agcgcgacgt
     4561 aatacgactc actatagggc gaattggcgg aaggccgtca aggccgtgac gttaaaacta
     4621 ttaagccatc caatcgaccg ttagtcgaat caggaccgct ggtgcgagaa gccgcgaagt
     4681 atggcgaatg catcgtataa cgtgtggagt ccgctcatta gagcgtcatg tttagacaag
     4741 aaagctacat atttaattga tcccgatgat tttattgata aattgaccct aactccatac
     4801 acggtattct acaatggcgg ggttttggtc aaaatttccg gactgcgatt gtacatgctg
     4861 ttaacggctc cgcccactat taatgaaatt aaaaattcca attttaaaaa acgcagcaag
     4921 agaaacattt gtatgaaaga atgcgtagaa ggaaagaaaa atgtcgtgga catgctgaac
     4981 aacaagatta atatgcctcc gtgtataaaa aaaatattga acgatttgaa agaaaacaat
     5041 gtaccgcgcg gcggtatgta caggaagagg tttatactaa actgttacat tgcaaacgtg
     5101 gtttcgtgtg ccaagtgtga aaaccgatgt ttaatcaagg ctctgacgca tttctacaac
     5161 cacgactcca agtgtgtggg tgaagtcatg catcttttaa tcaaatccca agatgtgtat
     5221 aaaccaccaa actgccaaaa aatgaaaact gtggacaagc tctgtccgtt tgctggcaac
     5281 tgcaagggtc tcaatcctat ttgtaattat tgaataataa aacaattata aatgctaaat
     5341 ttgtttttta ttaacgatac aaaccaaacg caacaagaac atttgtagta ttatctataa
     5401 ttgaaaacgc gtagttataa tcgctgaggt aatatttaaa atcattttca aatgattcac
     5461 agttaatttg cgacaatata attttatttt cacataaact agacgccttg tcgtcttctt
     5521 cttcgtattc cttctctttt tcatttttct cctcataaaa attaacatag ttattatcgt
     5581 atccatatat gtatctatcg tatagagtaa attttttgtt gtcataaata tatatgtctt
     5641 ttttaatggg gtgtatagta ccgctgcgca tagtttttct gtaatttaca acagtgctat
     5701 tttctggtag ttcttcggag tgtgttgctt taattattaa atttatataa tcaatgaatt
     5761 tgggatcgtc ggttttgtac aatatgttgc cggcatagta cgcagcttct tctagttcaa
     5821 ttacaccatt ttttagcagc accggattaa cataactttc caaaatgttg tacgaaccgt
     5881 taaacaaaaa cagttcacct cccttttcta tactattgtc tgcgagcagt tgtttgttgt
     5941 taaaaataac agccattgta atgagacgca caaactaata tcacaagctg ccaccatttt
     6001 gttcgcccag gactctagct atagttctag tggttggcta cgtatactcc ggaatattaa
     6061 tag
//
