LOCUS       sequence-224661-        6157 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 C-terminal mCitrine, cleavable with TEV.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6157)
  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 6157)
  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: pOCC230
FEATURES             Location/Qualifiers
     source          1..6157
                     /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             392..697
                     /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     576..595
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             753..773
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(774..776,777..779,780..1493)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY*"
     primer_bind     complement(819..840)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1080..1099)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1427..1448
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1623..1757
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1660..1679)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1714..1733
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1830..2202
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(2554..2571)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2664..3252)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2744..2763)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3423..4214,4215..4283))
                     /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     4046..4065
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4285..4389)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4690..4712
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4704..4721
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4705..4721
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4732..4751
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4732..4750
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5361..5485
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5523..6128)
                     /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 ccggcggccg caagcgctat
      301 ggccacaacc tgttacattg cacaagataa aaatatatca tcacgaacag taaaactgtc
      361 tgcttacata aacagtaata caaggggtgt tatgcagttt aaggtttaca cctataaaag
      421 agagagccgt tatcgtctgt ttgtggatgt acagagtgat attattgaca cgcccgggcg
      481 acggatggtg atccccctgg ccagtgcccg tctgctgtca gataaagtct cccgtgaact
      541 ttacccggtg gtgcatatcg gggatgaaag ctggcgcatg atgaccaccg atatggccag
      601 tgtgccggtc tccgttatcg gggaagaagt ggctgatctc agccaccgcg aaaatgacat
      661 caaaaacgcc attaacctga tgttctgggg aatataaatg tcaggctccc ttatacacag
      721 ccagtggcgc gccgggatcc agcagcggca gagaaaactt gtatttccag ggcatggtga
      781 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
      841 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
      901 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
      961 ccaccttcgg ctacggcctg atgtgcttcg cccgctaccc cgaccacatg aagcagcacg
     1021 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     1081 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     1141 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     1201 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
     1261 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     1321 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     1381 gctaccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     1441 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtactaa tagaagcttg
     1501 tcgagaagta ctagaggatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     1561 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     1621 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     1681 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     1741 cttatcatgt ctggatctga tcactgcttg agcctaggag atccgaacca gataagtgaa
     1801 agcctgcatc cgcagatctt aattaataaa acacgataca ttgttattag tacatttatt
     1861 aagcgctaga ttctgtgcgt tgttgattta cagacaattg ttgtacgtat tttaataatt
     1921 cattaaattt ataatcttta gggtggtatg ttagagcgaa aatcaaatga ttttcagcgt
     1981 ctttatatct gaatttaaat attaaatcct caatagattt gtaaaatagg tttcgattag
     2041 tttcaaacaa gggttgtttt tccgaaccga tggctggact atctaatgga ttttcgctca
     2101 acgccacaaa acttgccaaa tcttgtagca gcaatctagc tttgtcgata ttcgtttgtg
     2161 ttttgttttg taataaaggt tcgacgtcgt tcaaaatatt atgcgctttt gtatttcttt
     2221 catcactgtc gttagtgtac aattgactcg acgtaaacac gttaaataac gcttggacat
     2281 atttaacatc gggcgtgtta gctttattag gccgattatc gtcgtcgtcc caaccctcgt
     2341 cgttagaagt tgcttccgaa gacgattttg ccatagccac acgacgccta ttaattgtgt
     2401 cggctaacac gtccgcgatc aaatttgtag ttgagctttt tgggcctcat gggccttccg
     2461 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaac atggtcatag
     2521 ctgtttcctt gcgtattggg cgctctccgc ttcctcgctc actgactcgc tgcgctcggt
     2581 cgttcgggta aagcctgggg tgcctaatga gcaaaaggcc agcaaaaggc caggaaccgt
     2641 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     2701 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     2761 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     2821 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     2881 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     2941 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     3001 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     3061 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     3121 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     3181 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     3241 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     3301 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     3361 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     3421 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     3481 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     3541 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     3601 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     3661 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     3721 aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     3781 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     3841 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     3901 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     3961 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     4021 tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg
     4081 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     4141 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     4201 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     4261 acacggaaat gttgaatact cattactctt cctttttcaa tattattgaa gcatttatca
     4321 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     4381 ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta
     4441 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     4501 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtggccg ctacagggcg
     4561 ctcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgtttcggtg cgggcctctt
     4621 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
     4681 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgagcgcgac gtaatacgac
     4741 tcactatagg gcgaattggc ggaaggccgt caaggccgtg acgttaaaac tattaagcca
     4801 tccaatcgac cgttagtcga atcaggaccg ctggtgcgag aagccgcgaa gtatggcgaa
     4861 tgcatcgtat aacgtgtgga gtccgctcat tagagcgtca tgtttagaca agaaagctac
     4921 atatttaatt gatcccgatg attttattga taaattgacc ctaactccat acacggtatt
     4981 ctacaatggc ggggttttgg tcaaaatttc cggactgcga ttgtacatgc tgttaacggc
     5041 tccgcccact attaatgaaa ttaaaaattc caattttaaa aaacgcagca agagaaacat
     5101 ttgtatgaaa gaatgcgtag aaggaaagaa aaatgtcgtg gacatgctga acaacaagat
     5161 taatatgcct ccgtgtataa aaaaaatatt gaacgatttg aaagaaaaca atgtaccgcg
     5221 cggcggtatg tacaggaaga ggtttatact aaactgttac attgcaaacg tggtttcgtg
     5281 tgccaagtgt gaaaaccgat gtttaatcaa ggctctgacg catttctaca accacgactc
     5341 caagtgtgtg ggtgaagtca tgcatctttt aatcaaatcc caagatgtgt ataaaccacc
     5401 aaactgccaa aaaatgaaaa ctgtggacaa gctctgtccg tttgctggca actgcaaggg
     5461 tctcaatcct atttgtaatt attgaataat aaaacaatta taaatgctaa atttgttttt
     5521 tattaacgat acaaaccaaa cgcaacaaga acatttgtag tattatctat aattgaaaac
     5581 gcgtagttat aatcgctgag gtaatattta aaatcatttt caaatgattc acagttaatt
     5641 tgcgacaata taattttatt ttcacataaa ctagacgcct tgtcgtcttc ttcttcgtat
     5701 tccttctctt tttcattttt ctcctcataa aaattaacat agttattatc gtatccatat
     5761 atgtatctat cgtatagagt aaattttttg ttgtcataaa tatatatgtc ttttttaatg
     5821 gggtgtatag taccgctgcg catagttttt ctgtaattta caacagtgct attttctggt
     5881 agttcttcgg agtgtgttgc tttaattatt aaatttatat aatcaatgaa tttgggatcg
     5941 tcggttttgt acaatatgtt gccggcatag tacgcagctt cttctagttc aattacacca
     6001 ttttttagca gcaccggatt aacataactt tccaaaatgt tgtacgaacc gttaaacaaa
     6061 aacagttcac ctcccttttc tatactattg tctgcgagca gttgtttgtt gttaaaaata
     6121 acagccattg taatgagacg cacaaactaa tatcaca
//
