LOCUS       sequence-232359-        5982 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with C-terminal SNAP tag, cleavable with 
            TEV, and C-terminal HIS6 tag, cleavable with 3C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5982)
  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 5982)
  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: pOCC194
FEATURES             Location/Qualifiers
     source          1..5982
                     /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"
     regulatory      138..147
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             251..556
                     /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     435..454
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             612..632
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             633..1178
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     CDS             1179..1202
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1212..1229
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1371..1505
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1408..1427)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1462..1481
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1578..1950
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(2302..2319)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2412..3000)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2492..2511)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3171..3962,3963..4031))
                     /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     3794..3813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4032..4136)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4437..4459
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4451..4468
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4452..4468
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4479..4498
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4479..4497
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5108..5232
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5270..5875)
                     /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 tggcggccgc aagcgctatg gccacaacct gttacattgc
      181 acaagataaa aatatatcat cacgaacagt aaaactgtct gcttacataa acagtaatac
      241 aaggggtgtt atgcagttta aggtttacac ctataaaaga gagagccgtt atcgtctgtt
      301 tgtggatgta cagagtgata ttattgacac gcccgggcga cggatggtga tccccctggc
      361 cagtgcccgt ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg
      421 ggatgaaagc tggcgcatga tgaccaccga tatggccagt gtgccggtct ccgttatcgg
      481 ggaagaagtg gctgatctca gccaccgcga aaatgacatc aaaaacgcca ttaacctgat
      541 gttctgggga atataaatgt caggctccct tatacacagc cagtggcgcg ccgggatcca
      601 gcagcggcag agaaaacttg tatttccagg gcatggacaa agactgcgaa atgaagcgca
      661 ccaccctgga tagccctctg ggcaagctgg aactgtctgg gtgcgaacag ggcctgcacc
      721 gtatcatctt cctgggcaaa ggaacatctg ccgccgacgc cgtggaagtg cctgccccag
      781 ccgccgtgct gggcggacca gagccactga tgcaggccac cgcctggctc aacgcctact
      841 ttcaccagcc tgaggccatc gaggagttcc ctgtgccagc cctgcaccac ccagtgttcc
      901 agcaggagag ctttacccgc caggtgctgt ggaaactgct gaaagtggtg aagttcggag
      961 aggtcatcag ctacagccac ctggccgccc tggccggcaa tcccgccgcc accgccgccg
     1021 tgaaaaccgc cctgagcgga aatcccgtgc ccattctgat cccctgccac cgggtggtgc
     1081 agggcgacct ggacgtgggg ggctacgagg gcgggctcgc cgtgaaagag tggctgctgg
     1141 cccacgaggg ccacagactg ggcaagcctg ggctgggtct ggaagttctg ttccaggggc
     1201 cgggcagcag ccatcaccat caccatcaca gtggataata gaagcttgtc gagaagtact
     1261 agaggatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     1321 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     1381 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     1441 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     1501 ggatctgatc actgcttgag cctaggagat ccgaaccaga taagtgaaag cctgcatccg
     1561 cagatcttaa ttaataaaac acgatacatt gttattagta catttattaa gcgctagatt
     1621 ctgtgcgttg ttgatttaca gacaattgtt gtacgtattt taataattca ttaaatttat
     1681 aatctttagg gtggtatgtt agagcgaaaa tcaaatgatt ttcagcgtct ttatatctga
     1741 atttaaatat taaatcctca atagatttgt aaaataggtt tcgattagtt tcaaacaagg
     1801 gttgtttttc cgaaccgatg gctggactat ctaatggatt ttcgctcaac gccacaaaac
     1861 ttgccaaatc ttgtagcagc aatctagctt tgtcgatatt cgtttgtgtt ttgttttgta
     1921 ataaaggttc gacgtcgttc aaaatattat gcgcttttgt atttctttca tcactgtcgt
     1981 tagtgtacaa ttgactcgac gtaaacacgt taaataacgc ttggacatat ttaacatcgg
     2041 gcgtgttagc tttattaggc cgattatcgt cgtcgtccca accctcgtcg ttagaagttg
     2101 cttccgaaga cgattttgcc atagccacac gacgcctatt aattgtgtcg gctaacacgt
     2161 ccgcgatcaa atttgtagtt gagctttttg ggcctcatgg gccttccgct cactgcccgc
     2221 tttccagtcg ggaaacctgt cgtgccagct gcattaacat ggtcatagct gtttccttgc
     2281 gtattgggcg ctctccgctt cctcgctcac tgactcgctg cgctcggtcg ttcgggtaaa
     2341 gcctggggtg cctaatgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2401 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2461 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2521 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2581 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     2641 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     2701 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     2761 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     2821 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     2881 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     2941 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3001 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3061 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     3241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     3301 atgataccgc gagaaccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     3361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     3421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     3481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     3541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     3601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     3661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     3721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     3781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     3841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     3901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     3961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     4141 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     4201 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     4261 aatcaaaaga atagaccgag atagggttga gtggccgcta cagggcgctc ccattcgcca
     4321 ttcaggctgc gcaactgttg ggaagggcgt ttcggtgcgg gcctcttcgc tattacgcca
     4381 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     4441 gtcacgacgt tgtaaaacga cggccagtga gcgcgacgta atacgactca ctatagggcg
     4501 aattggcgga aggccgtcaa ggccgtgacg ttaaaactat taagccatcc aatcgaccgt
     4561 tagtcgaatc aggaccgctg gtgcgagaag ccgcgaagta tggcgaatgc atcgtataac
     4621 gtgtggagtc cgctcattag agcgtcatgt ttagacaaga aagctacata tttaattgat
     4681 cccgatgatt ttattgataa attgacccta actccataca cggtattcta caatggcggg
     4741 gttttggtca aaatttccgg actgcgattg tacatgctgt taacggctcc gcccactatt
     4801 aatgaaatta aaaattccaa ttttaaaaaa cgcagcaaga gaaacatttg tatgaaagaa
     4861 tgcgtagaag gaaagaaaaa tgtcgtggac atgctgaaca acaagattaa tatgcctccg
     4921 tgtataaaaa aaatattgaa cgatttgaaa gaaaacaatg taccgcgcgg cggtatgtac
     4981 aggaagaggt ttatactaaa ctgttacatt gcaaacgtgg tttcgtgtgc caagtgtgaa
     5041 aaccgatgtt taatcaaggc tctgacgcat ttctacaacc acgactccaa gtgtgtgggt
     5101 gaagtcatgc atcttttaat caaatcccaa gatgtgtata aaccaccaaa ctgccaaaaa
     5161 atgaaaactg tggacaagct ctgtccgttt gctggcaact gcaagggtct caatcctatt
     5221 tgtaattatt gaataataaa acaattataa atgctaaatt tgttttttat taacgataca
     5281 aaccaaacgc aacaagaaca tttgtagtat tatctataat tgaaaacgcg tagttataat
     5341 cgctgaggta atatttaaaa tcattttcaa atgattcaca gttaatttgc gacaatataa
     5401 ttttattttc acataaacta gacgccttgt cgtcttcttc ttcgtattcc ttctcttttt
     5461 catttttctc ctcataaaaa ttaacatagt tattatcgta tccatatatg tatctatcgt
     5521 atagagtaaa ttttttgttg tcataaatat atatgtcttt tttaatgggg tgtatagtac
     5581 cgctgcgcat agtttttctg taatttacaa cagtgctatt ttctggtagt tcttcggagt
     5641 gtgttgcttt aattattaaa tttatataat caatgaattt gggatcgtcg gttttgtaca
     5701 atatgttgcc ggcatagtac gcagcttctt ctagttcaat tacaccattt tttagcagca
     5761 ccggattaac ataactttcc aaaatgttgt acgaaccgtt aaacaaaaac agttcacctc
     5821 ccttttctat actattgtct gcgagcagtt gtttgttgtt aaaaataaca gccattgtaa
     5881 tgagacgcac aaactaatat cacaagctgc caccattttg ttcgcccagg actctagcta
     5941 tagttctagt ggttggctac gtatactccg gaatattaat ag
//
