LOCUS       sequence-232314-        5340 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for untagged recombinant protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5340)
  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 5340)
  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: pOCC82
FEATURES             Location/Qualifiers
     source          1..5340
                     /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"
     polyA_signal    729..863
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(766..785)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     820..839
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           936..1308
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(1660..1677)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1770..2358)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1850..1869)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2529..3320,3321..3389))
                     /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     3152..3171
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3390..3494)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3795..3817
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3809..3826
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3810..3826
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3837..3856
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3837..3855
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4466..4590
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(4628..5233)
                     /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 ccgtaataga
      601 agcttgtcga gaagtactag aggatcataa tcagccatac cacatttgta gaggttttac
      661 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
      721 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
      781 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
      841 atgtatctta tcatgtctgg atctgatcac tgcttgagcc taggagatcc gaaccagata
      901 agtgaaagcc tgcatccgca gatcttaatt aataaaacac gatacattgt tattagtaca
      961 tttattaagc gctagattct gtgcgttgtt gatttacaga caattgttgt acgtatttta
     1021 ataattcatt aaatttataa tctttagggt ggtatgttag agcgaaaatc aaatgatttt
     1081 cagcgtcttt atatctgaat ttaaatatta aatcctcaat agatttgtaa aataggtttc
     1141 gattagtttc aaacaagggt tgtttttccg aaccgatggc tggactatct aatggatttt
     1201 cgctcaacgc cacaaaactt gccaaatctt gtagcagcaa tctagctttg tcgatattcg
     1261 tttgtgtttt gttttgtaat aaaggttcga cgtcgttcaa aatattatgc gcttttgtat
     1321 ttctttcatc actgtcgtta gtgtacaatt gactcgacgt aaacacgtta aataacgctt
     1381 ggacatattt aacatcgggc gtgttagctt tattaggccg attatcgtcg tcgtcccaac
     1441 cctcgtcgtt agaagttgct tccgaagacg attttgccat agccacacga cgcctattaa
     1501 ttgtgtcggc taacacgtcc gcgatcaaat ttgtagttga gctttttggg cctcatgggc
     1561 cttccgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaacatgg
     1621 tcatagctgt ttccttgcgt attgggcgct ctccgcttcc tcgctcactg actcgctgcg
     1681 ctcggtcgtt cgggtaaagc ctggggtgcc taatgagcaa aaggccagca aaaggccagg
     1741 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     1801 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     1861 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     1921 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     1981 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2041 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2101 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2161 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     2221 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     2281 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     2341 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     2401 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     2461 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     2521 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     2581 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     2641 tctggcccca gtgctgcaat gataccgcga gaaccacgct caccggctcc agatttatca
     2701 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     2761 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     2821 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     2881 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     2941 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     3001 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     3061 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     3121 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     3181 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     3241 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     3301 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     3361 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     3421 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     3481 ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt
     3541 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     3601 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt ggccgctaca
     3661 gggcgctccc attcgccatt caggctgcgc aactgttggg aagggcgttt cggtgcgggc
     3721 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     3781 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgagc gcgacgtaat
     3841 acgactcact atagggcgaa ttggcggaag gccgtcaagg ccgtgacgtt aaaactatta
     3901 agccatccaa tcgaccgtta gtcgaatcag gaccgctggt gcgagaagcc gcgaagtatg
     3961 gcgaatgcat cgtataacgt gtggagtccg ctcattagag cgtcatgttt agacaagaaa
     4021 gctacatatt taattgatcc cgatgatttt attgataaat tgaccctaac tccatacacg
     4081 gtattctaca atggcggggt tttggtcaaa atttccggac tgcgattgta catgctgtta
     4141 acggctccgc ccactattaa tgaaattaaa aattccaatt ttaaaaaacg cagcaagaga
     4201 aacatttgta tgaaagaatg cgtagaagga aagaaaaatg tcgtggacat gctgaacaac
     4261 aagattaata tgcctccgtg tataaaaaaa atattgaacg atttgaaaga aaacaatgta
     4321 ccgcgcggcg gtatgtacag gaagaggttt atactaaact gttacattgc aaacgtggtt
     4381 tcgtgtgcca agtgtgaaaa ccgatgttta atcaaggctc tgacgcattt ctacaaccac
     4441 gactccaagt gtgtgggtga agtcatgcat cttttaatca aatcccaaga tgtgtataaa
     4501 ccaccaaact gccaaaaaat gaaaactgtg gacaagctct gtccgtttgc tggcaactgc
     4561 aagggtctca atcctatttg taattattga ataataaaac aattataaat gctaaatttg
     4621 ttttttatta acgatacaaa ccaaacgcaa caagaacatt tgtagtatta tctataattg
     4681 aaaacgcgta gttataatcg ctgaggtaat atttaaaatc attttcaaat gattcacagt
     4741 taatttgcga caatataatt ttattttcac ataaactaga cgccttgtcg tcttcttctt
     4801 cgtattcctt ctctttttca tttttctcct cataaaaatt aacatagtta ttatcgtatc
     4861 catatatgta tctatcgtat agagtaaatt ttttgttgtc ataaatatat atgtcttttt
     4921 taatggggtg tatagtaccg ctgcgcatag tttttctgta atttacaaca gtgctatttt
     4981 ctggtagttc ttcggagtgt gttgctttaa ttattaaatt tatataatca atgaatttgg
     5041 gatcgtcggt tttgtacaat atgttgccgg catagtacgc agcttcttct agttcaatta
     5101 caccattttt tagcagcacc ggattaacat aactttccaa aatgttgtac gaaccgttaa
     5161 acaaaaacag ttcacctccc ttttctatac tattgtctgc gagcagttgt ttgttgttaa
     5221 aaataacagc cattgtaatg agacgcacaa actaatatca caagctgcca ccattttgtt
     5281 cgcccaggac tctagctata gttctagtgg ttggctacgt atactccgga atattaatag
//
