LOCUS       sequence-224692-        5455 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with C-terminal TwinStrep, cleavable with 
            3C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5455)
  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 5455)
  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: pOCC154
FEATURES             Location/Qualifiers
     source          1..5455
                     /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             672..695
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(696..701,702..725,726..761,762..785)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="SAWSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     polyA_signal    921..1055
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(958..977)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1012..1031
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1128..1500
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(1852..1869)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1962..2550)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2042..2061)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2721..3512,3513..3581))
                     /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     3344..3363
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3583..3687)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3988..4010
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4002..4019
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4003..4019
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4030..4049
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4030..4048
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4659..4783
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(4821..5426)
                     /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 gctggaagtt ctgttccagg ggccgagcgc ttggagccat ccgcagtttg
      721 aaaaaggtgg tggtagcggt ggtggttcag gtggtagtgc atggtcacac cctcagtttg
      781 agaaataata gaagcttgtc gagaagtact agaggatcat aatcagccat accacatttg
      841 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
      901 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
      961 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     1021 ccaaactcat caatgtatct tatcatgtct ggatctgatc actgcttgag cctaggagat
     1081 ccgaaccaga taagtgaaag cctgcatccg cagatcttaa ttaataaaac acgatacatt
     1141 gttattagta catttattaa gcgctagatt ctgtgcgttg ttgatttaca gacaattgtt
     1201 gtacgtattt taataattca ttaaatttat aatctttagg gtggtatgtt agagcgaaaa
     1261 tcaaatgatt ttcagcgtct ttatatctga atttaaatat taaatcctca atagatttgt
     1321 aaaataggtt tcgattagtt tcaaacaagg gttgtttttc cgaaccgatg gctggactat
     1381 ctaatggatt ttcgctcaac gccacaaaac ttgccaaatc ttgtagcagc aatctagctt
     1441 tgtcgatatt cgtttgtgtt ttgttttgta ataaaggttc gacgtcgttc aaaatattat
     1501 gcgcttttgt atttctttca tcactgtcgt tagtgtacaa ttgactcgac gtaaacacgt
     1561 taaataacgc ttggacatat ttaacatcgg gcgtgttagc tttattaggc cgattatcgt
     1621 cgtcgtccca accctcgtcg ttagaagttg cttccgaaga cgattttgcc atagccacac
     1681 gacgcctatt aattgtgtcg gctaacacgt ccgcgatcaa atttgtagtt gagctttttg
     1741 ggcctcatgg gccttccgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     1801 gcattaacat ggtcatagct gtttccttgc gtattgggcg ctctccgctt cctcgctcac
     1861 tgactcgctg cgctcggtcg ttcgggtaaa gcctggggtg cctaatgagc aaaaggccag
     1921 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     1981 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2041 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2101 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2161 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2221 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2281 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2341 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2401 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2461 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2521 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2581 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     2641 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     2701 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     2761 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     2821 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     2881 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     2941 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3001 ccagttaata gtttgcgcaa cgttgttgcc attgctgcag gcatcgtggt gtcacgctcg
     3061 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     3121 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     3181 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3241 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     3301 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac gggataatac cgcgccacat
     3361 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     3421 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     3481 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     3541 aaaaagggaa taagggcgac acggaaatgt tgaatactca ttactcttcc tttttcaata
     3601 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     3661 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta
     3721 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     3781 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     3841 agtggccgct acagggcgct cccattcgcc attcaggctg cgcaactgtt gggaagggcg
     3901 tttcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     3961 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     4021 agcgcgacgt aatacgactc actatagggc gaattggcgg aaggccgtca aggccgtgac
     4081 gttaaaacta ttaagccatc caatcgaccg ttagtcgaat caggaccgct ggtgcgagaa
     4141 gccgcgaagt atggcgaatg catcgtataa cgtgtggagt ccgctcatta gagcgtcatg
     4201 tttagacaag aaagctacat atttaattga tcccgatgat tttattgata aattgaccct
     4261 aactccatac acggtattct acaatggcgg ggttttggtc aaaatttccg gactgcgatt
     4321 gtacatgctg ttaacggctc cgcccactat taatgaaatt aaaaattcca attttaaaaa
     4381 acgcagcaag agaaacattt gtatgaaaga atgcgtagaa ggaaagaaaa atgtcgtgga
     4441 catgctgaac aacaagatta atatgcctcc gtgtataaaa aaaatattga acgatttgaa
     4501 agaaaacaat gtaccgcgcg gcggtatgta caggaagagg tttatactaa actgttacat
     4561 tgcaaacgtg gtttcgtgtg ccaagtgtga aaaccgatgt ttaatcaagg ctctgacgca
     4621 tttctacaac cacgactcca agtgtgtggg tgaagtcatg catcttttaa tcaaatccca
     4681 agatgtgtat aaaccaccaa actgccaaaa aatgaaaact gtggacaagc tctgtccgtt
     4741 tgctggcaac tgcaagggtc tcaatcctat ttgtaattat tgaataataa aacaattata
     4801 aatgctaaat ttgtttttta ttaacgatac aaaccaaacg caacaagaac atttgtagta
     4861 ttatctataa ttgaaaacgc gtagttataa tcgctgaggt aatatttaaa atcattttca
     4921 aatgattcac agttaatttg cgacaatata attttatttt cacataaact agacgccttg
     4981 tcgtcttctt cttcgtattc cttctctttt tcatttttct cctcataaaa attaacatag
     5041 ttattatcgt atccatatat gtatctatcg tatagagtaa attttttgtt gtcataaata
     5101 tatatgtctt ttttaatggg gtgtatagta ccgctgcgca tagtttttct gtaatttaca
     5161 acagtgctat tttctggtag ttcttcggag tgtgttgctt taattattaa atttatataa
     5221 tcaatgaatt tgggatcgtc ggttttgtac aatatgttgc cggcatagta cgcagcttct
     5281 tctagttcaa ttacaccatt ttttagcagc accggattaa cataactttc caaaatgttg
     5341 tacgaaccgt taaacaaaaa cagttcacct cccttttcta tactattgtc tgcgagcagt
     5401 tgtttgttgt taaaaataac agccattgta atgagacgca caaactaata tcaca
//
