LOCUS       sequence-224696-        5389 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with C-terminal sortase site (LPETGg) and 
            HIS6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5389)
  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 5389)
  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: pOCC111
FEATURES             Location/Qualifiers
     source          1..5389
                     /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"
     regulatory      216..225
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             329..634
                     /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     513..532
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             696..713
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    855..989
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(892..911)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     946..965
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1062..1434
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(1786..1803)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1896..2484)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1976..1995)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2655..3446,3447..3515))
                     /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     3278..3297
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3517..3621)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3922..3944
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3936..3953
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3937..3953
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3964..3983
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3964..3982
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4593..4717
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(4755..5360)
                     /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 gcggccgcaa gcgctatggc
      241 cacaacctgt tacattgcac aagataaaaa tatatcatca cgaacagtaa aactgtctgc
      301 ttacataaac agtaatacaa ggggtgttat gcagtttaag gtttacacct ataaaagaga
      361 gagccgttat cgtctgtttg tggatgtaca gagtgatatt attgacacgc ccgggcgacg
      421 gatggtgatc cccctggcca gtgcccgtct gctgtcagat aaagtctccc gtgaacttta
      481 cccggtggtg catatcgggg atgaaagctg gcgcatgatg accaccgata tggccagtgt
      541 gccggtctcc gttatcgggg aagaagtggc tgatctcagc caccgcgaaa atgacatcaa
      601 aaacgccatt aacctgatgt tctggggaat ataaatgtca ggctccctta tacacagcca
      661 gtggcgcgcc gctgcccgag accggcggca gcagccatca ccatcaccat cacagtggat
      721 aatagaagct tgtcgagaag tactagagga tcataatcag ccataccaca tttgtagagg
      781 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
      841 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
      901 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
      961 tcatcaatgt atcttatcat gtctggatct gatcactgct tgagcctagg agatccgaac
     1021 cagataagtg aaagcctgca tccgcagatc ttaattaata aaacacgata cattgttatt
     1081 agtacattta ttaagcgcta gattctgtgc gttgttgatt tacagacaat tgttgtacgt
     1141 attttaataa ttcattaaat ttataatctt tagggtggta tgttagagcg aaaatcaaat
     1201 gattttcagc gtctttatat ctgaatttaa atattaaatc ctcaatagat ttgtaaaata
     1261 ggtttcgatt agtttcaaac aagggttgtt tttccgaacc gatggctgga ctatctaatg
     1321 gattttcgct caacgccaca aaacttgcca aatcttgtag cagcaatcta gctttgtcga
     1381 tattcgtttg tgttttgttt tgtaataaag gttcgacgtc gttcaaaata ttatgcgctt
     1441 ttgtatttct ttcatcactg tcgttagtgt acaattgact cgacgtaaac acgttaaata
     1501 acgcttggac atatttaaca tcgggcgtgt tagctttatt aggccgatta tcgtcgtcgt
     1561 cccaaccctc gtcgttagaa gttgcttccg aagacgattt tgccatagcc acacgacgcc
     1621 tattaattgt gtcggctaac acgtccgcga tcaaatttgt agttgagctt tttgggcctc
     1681 atgggccttc cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     1741 acatggtcat agctgtttcc ttgcgtattg ggcgctctcc gcttcctcgc tcactgactc
     1801 gctgcgctcg gtcgttcggg taaagcctgg ggtgcctaat gagcaaaagg ccagcaaaag
     1861 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     1921 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     1981 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     2041 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     2101 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     2161 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     2221 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     2281 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     2341 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     2401 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     2461 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     2521 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     2581 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     2641 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     2701 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     2761 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     2821 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     2881 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     2941 aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt
     3001 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     3061 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     3121 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     3181 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     3241 cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga
     3301 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     3361 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     3421 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     3481 ggaataaggg cgacacggaa atgttgaata ctcattactc ttcctttttc aatattattg
     3541 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     3601 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt
     3661 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     3721 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtggc
     3781 cgctacaggg cgctcccatt cgccattcag gctgcgcaac tgttgggaag ggcgtttcgg
     3841 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     3901 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg
     3961 acgtaatacg actcactata gggcgaattg gcggaaggcc gtcaaggccg tgacgttaaa
     4021 actattaagc catccaatcg accgttagtc gaatcaggac cgctggtgcg agaagccgcg
     4081 aagtatggcg aatgcatcgt ataacgtgtg gagtccgctc attagagcgt catgtttaga
     4141 caagaaagct acatatttaa ttgatcccga tgattttatt gataaattga ccctaactcc
     4201 atacacggta ttctacaatg gcggggtttt ggtcaaaatt tccggactgc gattgtacat
     4261 gctgttaacg gctccgccca ctattaatga aattaaaaat tccaatttta aaaaacgcag
     4321 caagagaaac atttgtatga aagaatgcgt agaaggaaag aaaaatgtcg tggacatgct
     4381 gaacaacaag attaatatgc ctccgtgtat aaaaaaaata ttgaacgatt tgaaagaaaa
     4441 caatgtaccg cgcggcggta tgtacaggaa gaggtttata ctaaactgtt acattgcaaa
     4501 cgtggtttcg tgtgccaagt gtgaaaaccg atgtttaatc aaggctctga cgcatttcta
     4561 caaccacgac tccaagtgtg tgggtgaagt catgcatctt ttaatcaaat cccaagatgt
     4621 gtataaacca ccaaactgcc aaaaaatgaa aactgtggac aagctctgtc cgtttgctgg
     4681 caactgcaag ggtctcaatc ctatttgtaa ttattgaata ataaaacaat tataaatgct
     4741 aaatttgttt tttattaacg atacaaacca aacgcaacaa gaacatttgt agtattatct
     4801 ataattgaaa acgcgtagtt ataatcgctg aggtaatatt taaaatcatt ttcaaatgat
     4861 tcacagttaa tttgcgacaa tataatttta ttttcacata aactagacgc cttgtcgtct
     4921 tcttcttcgt attccttctc tttttcattt ttctcctcat aaaaattaac atagttatta
     4981 tcgtatccat atatgtatct atcgtataga gtaaattttt tgttgtcata aatatatatg
     5041 tcttttttaa tggggtgtat agtaccgctg cgcatagttt ttctgtaatt tacaacagtg
     5101 ctattttctg gtagttcttc ggagtgtgtt gctttaatta ttaaatttat ataatcaatg
     5161 aatttgggat cgtcggtttt gtacaatatg ttgccggcat agtacgcagc ttcttctagt
     5221 tcaattacac cattttttag cagcaccgga ttaacataac tttccaaaat gttgtacgaa
     5281 ccgttaaaca aaaacagttc acctcccttt tctatactat tgtctgcgag cagttgtttg
     5341 ttgttaaaaa taacagccat tgtaatgaga cgcacaaact aatatcaca
//
