LOCUS       sequence-232237-        5397 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with C-terminal HIS6 tag, cleavable with 3C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5397)
  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 5397)
  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: pOCC7
FEATURES             Location/Qualifiers
     source          1..5397
                     /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             594..617
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             627..644
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    786..920
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(823..842)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     877..896
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           993..1365
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(1717..1734)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1827..2415)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1907..1926)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2586..3377,3378..3446))
                     /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     3209..3228
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3447..3551)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3852..3874
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3866..3883
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3867..3883
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3894..3913
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3894..3912
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4523..4647
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(4685..5290)
                     /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 ccgctggaag
      601 ttctgttcca ggggccgggc agcagccatc accatcacca tcacagtgga taatagaagc
      661 ttgtcgagaa gtactagagg atcataatca gccataccac atttgtagag gttttacttg
      721 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
      781 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
      841 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
      901 tatcttatca tgtctggatc tgatcactgc ttgagcctag gagatccgaa ccagataagt
      961 gaaagcctgc atccgcagat cttaattaat aaaacacgat acattgttat tagtacattt
     1021 attaagcgct agattctgtg cgttgttgat ttacagacaa ttgttgtacg tattttaata
     1081 attcattaaa tttataatct ttagggtggt atgttagagc gaaaatcaaa tgattttcag
     1141 cgtctttata tctgaattta aatattaaat cctcaataga tttgtaaaat aggtttcgat
     1201 tagtttcaaa caagggttgt ttttccgaac cgatggctgg actatctaat ggattttcgc
     1261 tcaacgccac aaaacttgcc aaatcttgta gcagcaatct agctttgtcg atattcgttt
     1321 gtgttttgtt ttgtaataaa ggttcgacgt cgttcaaaat attatgcgct tttgtatttc
     1381 tttcatcact gtcgttagtg tacaattgac tcgacgtaaa cacgttaaat aacgcttgga
     1441 catatttaac atcgggcgtg ttagctttat taggccgatt atcgtcgtcg tcccaaccct
     1501 cgtcgttaga agttgcttcc gaagacgatt ttgccatagc cacacgacgc ctattaattg
     1561 tgtcggctaa cacgtccgcg atcaaatttg tagttgagct ttttgggcct catgggcctt
     1621 ccgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aacatggtca
     1681 tagctgtttc cttgcgtatt gggcgctctc cgcttcctcg ctcactgact cgctgcgctc
     1741 ggtcgttcgg gtaaagcctg gggtgcctaa tgagcaaaag gccagcaaaa ggccaggaac
     1801 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     1861 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     1921 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     1981 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     2041 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     2101 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     2161 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     2221 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     2281 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     2341 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     2401 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     2461 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     2521 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     2581 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     2641 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     2701 ggccccagtg ctgcaatgat accgcgagaa ccacgctcac cggctccaga tttatcagca
     2761 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     2821 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     2881 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     2941 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     3001 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     3061 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     3121 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     3181 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     3241 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     3301 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     3361 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     3421 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     3481 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     3541 ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt
     3601 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     3661 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtggc cgctacaggg
     3721 cgctcccatt cgccattcag gctgcgcaac tgttgggaag ggcgtttcgg tgcgggcctc
     3781 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     3841 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg acgtaatacg
     3901 actcactata gggcgaattg gcggaaggcc gtcaaggccg tgacgttaaa actattaagc
     3961 catccaatcg accgttagtc gaatcaggac cgctggtgcg agaagccgcg aagtatggcg
     4021 aatgcatcgt ataacgtgtg gagtccgctc attagagcgt catgtttaga caagaaagct
     4081 acatatttaa ttgatcccga tgattttatt gataaattga ccctaactcc atacacggta
     4141 ttctacaatg gcggggtttt ggtcaaaatt tccggactgc gattgtacat gctgttaacg
     4201 gctccgccca ctattaatga aattaaaaat tccaatttta aaaaacgcag caagagaaac
     4261 atttgtatga aagaatgcgt agaaggaaag aaaaatgtcg tggacatgct gaacaacaag
     4321 attaatatgc ctccgtgtat aaaaaaaata ttgaacgatt tgaaagaaaa caatgtaccg
     4381 cgcggcggta tgtacaggaa gaggtttata ctaaactgtt acattgcaaa cgtggtttcg
     4441 tgtgccaagt gtgaaaaccg atgtttaatc aaggctctga cgcatttcta caaccacgac
     4501 tccaagtgtg tgggtgaagt catgcatctt ttaatcaaat cccaagatgt gtataaacca
     4561 ccaaactgcc aaaaaatgaa aactgtggac aagctctgtc cgtttgctgg caactgcaag
     4621 ggtctcaatc ctatttgtaa ttattgaata ataaaacaat tataaatgct aaatttgttt
     4681 tttattaacg atacaaacca aacgcaacaa gaacatttgt agtattatct ataattgaaa
     4741 acgcgtagtt ataatcgctg aggtaatatt taaaatcatt ttcaaatgat tcacagttaa
     4801 tttgcgacaa tataatttta ttttcacata aactagacgc cttgtcgtct tcttcttcgt
     4861 attccttctc tttttcattt ttctcctcat aaaaattaac atagttatta tcgtatccat
     4921 atatgtatct atcgtataga gtaaattttt tgttgtcata aatatatatg tcttttttaa
     4981 tggggtgtat agtaccgctg cgcatagttt ttctgtaatt tacaacagtg ctattttctg
     5041 gtagttcttc ggagtgtgtt gctttaatta ttaaatttat ataatcaatg aatttgggat
     5101 cgtcggtttt gtacaatatg ttgccggcat agtacgcagc ttcttctagt tcaattacac
     5161 cattttttag cagcaccgga ttaacataac tttccaaaat gttgtacgaa ccgttaaaca
     5221 aaaacagttc acctcccttt tctatactat tgtctgcgag cagttgtttg ttgttaaaaa
     5281 taacagccat tgtaatgaga cgcacaaact aatatcacaa gctgccacca ttttgttcgc
     5341 ccaggactct agctatagtt ctagtggttg gctacgtata ctccggaata ttaatag
//
