LOCUS       sequence-232264-        6117 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with C-terminal Dendra2, and C-terminal HIS6
            tag, cleavable with 3C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6117)
  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 6117)
  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: pOCC215
FEATURES             Location/Qualifiers
     source          1..6117
                     /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             624..1313
                     /codon_start=1
                     /product="monomeric photoswitchable green-to-red 
                     fluorescent protein from Dendronephthya"
                     /label=Dendra2
                     /note="mammalian codon-optimized"
                     /translation="MNTPGINLIKEDMRVKVHMEGNVNGHAFVIEGEGKGKPYEGTQTA
                     NLTVKEGAPLPFSYDILTTAVHYGNRVFTKYPEDIPDYFKQSFPEGYSWERTMTFEDKG
                     ICTIRSDISLEGDCFFQNVRFKGTNFPPNGPVMQKKTLKWEPSTEKLHVRDGLLVGNIN
                     MALLLEGGGHYLCDFKTTYKAKKVVQLPDAHFVDHRIEILGNDSDYNKVKLYEHAVARY
                     SPLPSQVW"
     CDS             1314..1337
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1347..1364
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1506..1640
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1543..1562)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1597..1616
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1713..2085
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(2437..2454)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2547..3135)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2627..2646)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3306..4097,4098..4166))
                     /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     3929..3948
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4167..4271)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4572..4594
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4586..4603
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4587..4603
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4614..4633
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4614..4632
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5243..5367
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5405..6010)
                     /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 ccgggatccg
      601 ctggctccgc tgctggttct ggcatgaaca ccccgggaat taacctgatc aaggaggaca
      661 tgcgcgtgaa ggtgcacatg gagggcaacg tgaacggcca cgccttcgtg atcgagggcg
      721 agggcaaggg caagccctac gagggcaccc agaccgccaa cctgaccgtg aaggagggcg
      781 cccccctgcc cttcagctac gacatcctga ccaccgccgt gcactacggc aaccgggtgt
      841 tcaccaagta ccccgaggac atccccgact acttcaagca gagcttcccc gagggctaca
      901 gctgggagcg caccatgacc ttcgaggaca agggcatctg caccatccgc agcgacatca
      961 gcctggaggg cgactgcttc ttccagaacg tgcgcttcaa gggcaccaac ttccccccca
     1021 acggccccgt gatgcagaag aagaccctga agtgggagcc cagcaccgag aagctgcacg
     1081 tgcgcgacgg cctgctggtg ggcaacatca acatggccct gctgctggag ggcggcggcc
     1141 actacctgtg cgacttcaag accacctaca aggccaagaa ggtggtgcag ctgcccgacg
     1201 cccacttcgt ggaccaccgc atcgagatcc tgggcaacga cagcgactac aacaaggtga
     1261 agctgtacga gcacgccgtg gcccgctaca gccccctgcc cagccaggtg tggctggaag
     1321 ttctgttcca ggggccgggc agcagccatc accatcacca tcacagtgga taatagaagc
     1381 ttgtcgagaa gtactagagg atcataatca gccataccac atttgtagag gttttacttg
     1441 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     1501 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     1561 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     1621 tatcttatca tgtctggatc tgatcactgc ttgagcctag gagatccgaa ccagataagt
     1681 gaaagcctgc atccgcagat cttaattaat aaaacacgat acattgttat tagtacattt
     1741 attaagcgct agattctgtg cgttgttgat ttacagacaa ttgttgtacg tattttaata
     1801 attcattaaa tttataatct ttagggtggt atgttagagc gaaaatcaaa tgattttcag
     1861 cgtctttata tctgaattta aatattaaat cctcaataga tttgtaaaat aggtttcgat
     1921 tagtttcaaa caagggttgt ttttccgaac cgatggctgg actatctaat ggattttcgc
     1981 tcaacgccac aaaacttgcc aaatcttgta gcagcaatct agctttgtcg atattcgttt
     2041 gtgttttgtt ttgtaataaa ggttcgacgt cgttcaaaat attatgcgct tttgtatttc
     2101 tttcatcact gtcgttagtg tacaattgac tcgacgtaaa cacgttaaat aacgcttgga
     2161 catatttaac atcgggcgtg ttagctttat taggccgatt atcgtcgtcg tcccaaccct
     2221 cgtcgttaga agttgcttcc gaagacgatt ttgccatagc cacacgacgc ctattaattg
     2281 tgtcggctaa cacgtccgcg atcaaatttg tagttgagct ttttgggcct catgggcctt
     2341 ccgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aacatggtca
     2401 tagctgtttc cttgcgtatt gggcgctctc cgcttcctcg ctcactgact cgctgcgctc
     2461 ggtcgttcgg gtaaagcctg gggtgcctaa tgagcaaaag gccagcaaaa ggccaggaac
     2521 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     2581 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     2641 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     2701 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     2761 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     2821 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     2881 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     2941 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     3001 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3061 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3121 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3181 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3241 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     3301 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     3361 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     3421 ggccccagtg ctgcaatgat accgcgagaa ccacgctcac cggctccaga tttatcagca
     3481 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     3541 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     3601 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     3661 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     3721 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     3781 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     3841 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     3901 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     3961 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4021 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4081 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4141 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     4201 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     4261 ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt
     4321 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     4381 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtggc cgctacaggg
     4441 cgctcccatt cgccattcag gctgcgcaac tgttgggaag ggcgtttcgg tgcgggcctc
     4501 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     4561 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg acgtaatacg
     4621 actcactata gggcgaattg gcggaaggcc gtcaaggccg tgacgttaaa actattaagc
     4681 catccaatcg accgttagtc gaatcaggac cgctggtgcg agaagccgcg aagtatggcg
     4741 aatgcatcgt ataacgtgtg gagtccgctc attagagcgt catgtttaga caagaaagct
     4801 acatatttaa ttgatcccga tgattttatt gataaattga ccctaactcc atacacggta
     4861 ttctacaatg gcggggtttt ggtcaaaatt tccggactgc gattgtacat gctgttaacg
     4921 gctccgccca ctattaatga aattaaaaat tccaatttta aaaaacgcag caagagaaac
     4981 atttgtatga aagaatgcgt agaaggaaag aaaaatgtcg tggacatgct gaacaacaag
     5041 attaatatgc ctccgtgtat aaaaaaaata ttgaacgatt tgaaagaaaa caatgtaccg
     5101 cgcggcggta tgtacaggaa gaggtttata ctaaactgtt acattgcaaa cgtggtttcg
     5161 tgtgccaagt gtgaaaaccg atgtttaatc aaggctctga cgcatttcta caaccacgac
     5221 tccaagtgtg tgggtgaagt catgcatctt ttaatcaaat cccaagatgt gtataaacca
     5281 ccaaactgcc aaaaaatgaa aactgtggac aagctctgtc cgtttgctgg caactgcaag
     5341 ggtctcaatc ctatttgtaa ttattgaata ataaaacaat tataaatgct aaatttgttt
     5401 tttattaacg atacaaacca aacgcaacaa gaacatttgt agtattatct ataattgaaa
     5461 acgcgtagtt ataatcgctg aggtaatatt taaaatcatt ttcaaatgat tcacagttaa
     5521 tttgcgacaa tataatttta ttttcacata aactagacgc cttgtcgtct tcttcttcgt
     5581 attccttctc tttttcattt ttctcctcat aaaaattaac atagttatta tcgtatccat
     5641 atatgtatct atcgtataga gtaaattttt tgttgtcata aatatatatg tcttttttaa
     5701 tggggtgtat agtaccgctg cgcatagttt ttctgtaatt tacaacagtg ctattttctg
     5761 gtagttcttc ggagtgtgtt gctttaatta ttaaatttat ataatcaatg aatttgggat
     5821 cgtcggtttt gtacaatatg ttgccggcat agtacgcagc ttcttctagt tcaattacac
     5881 cattttttag cagcaccgga ttaacataac tttccaaaat gttgtacgaa ccgttaaaca
     5941 aaaacagttc acctcccttt tctatactat tgtctgcgag cagttgtttg ttgttaaaaa
     6001 taacagccat tgtaatgaga cgcacaaact aatatcacaa gctgccacca ttttgttcgc
     6061 ccaggactct agctatagtt ctagtggttg gctacgtata ctccggaata ttaatag
//
