LOCUS       sequence-224658-        6127 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with N-terminal HIS6 tag, cleavable with 3C,
            and N-terminal tagRFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6127)
  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 6127)
  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: pOCC18
FEATURES             Location/Qualifiers
     source          1..6127
                     /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             284..973
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSDMALK
                     LVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAVARYC
                     DLPSKLGH"
     CDS             1115..1420
                     /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     1299..1318
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    1593..1727
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1630..1649)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1684..1703
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1800..2172
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(2524..2541)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2634..3222)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2714..2733)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3393..4184,4185..4253))
                     /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     4016..4035
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4255..4359)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4660..4682
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4674..4691
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4675..4691
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4702..4721
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4702..4720
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5331..5455
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5493..6098)
                     /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 ccgatgagcg agctgattaa
      301 ggagaacatg cacatgaagc tgtacatgga gggcaccgtg aacaaccacc acttcaagtg
      361 cacatccgag ggcgaaggca agccctacga gggcacccag accatgagaa tcaaggtggt
      421 cgagggcggc cctctcccct tcgccttcga catcctggct accagcttca tgtacggcag
      481 cagaaccttc atcaaccaca cccagggcat ccccgacttc tttaagcagt ccttccctga
      541 gggcttcaca tgggagagag tcaccacata cgaagacggg ggcgtgctga ccgctaccca
      601 ggacaccagc ctccaggacg gctgcctcat ctacaacgtc aagatcagag gggtgaactt
      661 cccatccaac ggccctgtga tgcagaagaa aacactcggc tgggaggcca acaccgagat
      721 gctgtacccc gctgacggcg gcctggaagg cagaagcgac atggccctga agctcgtggg
      781 cgggggccac ctgatctgca acttcaagac cacatacaga tccaagaaac ccgctaagaa
      841 cctcaagatg cccggcgtct actatgtgga ccacagactg gaaagaatca aggaggccga
      901 caaagagacc tacgtcgagc agcacgaggt ggctgtggcc agatactgcg acctccctag
      961 caaactgggg cacaagggat ccgctggctc cgctgctggt tctggcgcgg ccgcaagcgc
     1021 tatggccaca acctgttaca ttgcacaaga taaaaatata tcatcacgaa cagtaaaact
     1081 gtctgcttac ataaacagta atacaagggg tgttatgcag tttaaggttt acacctataa
     1141 aagagagagc cgttatcgtc tgtttgtgga tgtacagagt gatattattg acacgcccgg
     1201 gcgacggatg gtgatccccc tggccagtgc ccgtctgctg tcagataaag tctcccgtga
     1261 actttacccg gtggtgcata tcggggatga aagctggcgc atgatgacca ccgatatggc
     1321 cagtgtgccg gtctccgtta tcggggaaga agtggctgat ctcagccacc gcgaaaatga
     1381 catcaaaaac gccattaacc tgatgttctg gggaatataa atgtcaggct cccttataca
     1441 cagccagtgg cgcgccgtaa tagaagcttg tcgagaagta ctagaggatc ataatcagcc
     1501 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     1561 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     1621 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     1681 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatctga tcactgcttg
     1741 agcctaggag atccgaacca gataagtgaa agcctgcatc cgcagatctt aattaataaa
     1801 acacgataca ttgttattag tacatttatt aagcgctaga ttctgtgcgt tgttgattta
     1861 cagacaattg ttgtacgtat tttaataatt cattaaattt ataatcttta gggtggtatg
     1921 ttagagcgaa aatcaaatga ttttcagcgt ctttatatct gaatttaaat attaaatcct
     1981 caatagattt gtaaaatagg tttcgattag tttcaaacaa gggttgtttt tccgaaccga
     2041 tggctggact atctaatgga ttttcgctca acgccacaaa acttgccaaa tcttgtagca
     2101 gcaatctagc tttgtcgata ttcgtttgtg ttttgttttg taataaaggt tcgacgtcgt
     2161 tcaaaatatt atgcgctttt gtatttcttt catcactgtc gttagtgtac aattgactcg
     2221 acgtaaacac gttaaataac gcttggacat atttaacatc gggcgtgtta gctttattag
     2281 gccgattatc gtcgtcgtcc caaccctcgt cgttagaagt tgcttccgaa gacgattttg
     2341 ccatagccac acgacgccta ttaattgtgt cggctaacac gtccgcgatc aaatttgtag
     2401 ttgagctttt tgggcctcat gggccttccg ctcactgccc gctttccagt cgggaaacct
     2461 gtcgtgccag ctgcattaac atggtcatag ctgtttcctt gcgtattggg cgctctccgc
     2521 ttcctcgctc actgactcgc tgcgctcggt cgttcgggta aagcctgggg tgcctaatga
     2581 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2641 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2701 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2761 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2821 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2881 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2941 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3001 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3061 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3121 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3181 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3241 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     3301 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     3361 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     3421 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     3481 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     3541 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     3601 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     3661 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg
     3721 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3781 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     3841 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     3901 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     3961 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat
     4021 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     4081 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     4141 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     4201 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact cattactctt
     4261 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4321 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     4381 acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     4441 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     4501 gagatagggt tgagtggccg ctacagggcg ctcccattcg ccattcaggc tgcgcaactg
     4561 ttgggaaggg cgtttcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
     4621 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
     4681 cgacggccag tgagcgcgac gtaatacgac tcactatagg gcgaattggc ggaaggccgt
     4741 caaggccgtg acgttaaaac tattaagcca tccaatcgac cgttagtcga atcaggaccg
     4801 ctggtgcgag aagccgcgaa gtatggcgaa tgcatcgtat aacgtgtgga gtccgctcat
     4861 tagagcgtca tgtttagaca agaaagctac atatttaatt gatcccgatg attttattga
     4921 taaattgacc ctaactccat acacggtatt ctacaatggc ggggttttgg tcaaaatttc
     4981 cggactgcga ttgtacatgc tgttaacggc tccgcccact attaatgaaa ttaaaaattc
     5041 caattttaaa aaacgcagca agagaaacat ttgtatgaaa gaatgcgtag aaggaaagaa
     5101 aaatgtcgtg gacatgctga acaacaagat taatatgcct ccgtgtataa aaaaaatatt
     5161 gaacgatttg aaagaaaaca atgtaccgcg cggcggtatg tacaggaaga ggtttatact
     5221 aaactgttac attgcaaacg tggtttcgtg tgccaagtgt gaaaaccgat gtttaatcaa
     5281 ggctctgacg catttctaca accacgactc caagtgtgtg ggtgaagtca tgcatctttt
     5341 aatcaaatcc caagatgtgt ataaaccacc aaactgccaa aaaatgaaaa ctgtggacaa
     5401 gctctgtccg tttgctggca actgcaaggg tctcaatcct atttgtaatt attgaataat
     5461 aaaacaatta taaatgctaa atttgttttt tattaacgat acaaaccaaa cgcaacaaga
     5521 acatttgtag tattatctat aattgaaaac gcgtagttat aatcgctgag gtaatattta
     5581 aaatcatttt caaatgattc acagttaatt tgcgacaata taattttatt ttcacataaa
     5641 ctagacgcct tgtcgtcttc ttcttcgtat tccttctctt tttcattttt ctcctcataa
     5701 aaattaacat agttattatc gtatccatat atgtatctat cgtatagagt aaattttttg
     5761 ttgtcataaa tatatatgtc ttttttaatg gggtgtatag taccgctgcg catagttttt
     5821 ctgtaattta caacagtgct attttctggt agttcttcgg agtgtgttgc tttaattatt
     5881 aaatttatat aatcaatgaa tttgggatcg tcggttttgt acaatatgtt gccggcatag
     5941 tacgcagctt cttctagttc aattacacca ttttttagca gcaccggatt aacataactt
     6001 tccaaaatgt tgtacgaacc gttaaacaaa aacagttcac ctcccttttc tatactattg
     6061 tctgcgagca gttgtttgtt gttaaaaata acagccattg taatgagacg cacaaactaa
     6121 tatcaca
//
