LOCUS       sequence-224667-        6145 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with C-terminal monomeric GFP and C-terminal
            HIS6 tag, cleavable with 3C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6145)
  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 6145)
  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: pOCC61
FEATURES             Location/Qualifiers
     source          1..6145
                     /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             join(702..704,705..707,708..1418)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(747..768)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1008..1027)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1355..1376
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1419..1442
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1452..1469
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1611..1745
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1648..1667)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1702..1721
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1818..2190
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(2542..2559)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2652..3240)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2732..2751)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3411..4202,4203..4271))
                     /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     4034..4053
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4273..4377)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4678..4700
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4692..4709
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4693..4709
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4720..4739
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4720..4738
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5349..5473
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5511..6116)
                     /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 gggatccgct ggctccgctg ctggttctgg catggtgagc aagggcgagg
      721 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
      781 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
      841 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
      901 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
      961 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     1021 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     1081 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     1141 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     1201 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     1261 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtcca
     1321 aactgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     1381 ccgccgggat cactctcggc atggacgagc tgtacaagct ggaagttctg ttccaggggc
     1441 cgggcagcag ccatcaccat caccatcaca gtggataata gaagcttgtc gagaagtact
     1501 agaggatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     1561 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     1621 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     1681 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     1741 ggatctgatc actgcttgag cctaggagat ccgaaccaga taagtgaaag cctgcatccg
     1801 cagatcttaa ttaataaaac acgatacatt gttattagta catttattaa gcgctagatt
     1861 ctgtgcgttg ttgatttaca gacaattgtt gtacgtattt taataattca ttaaatttat
     1921 aatctttagg gtggtatgtt agagcgaaaa tcaaatgatt ttcagcgtct ttatatctga
     1981 atttaaatat taaatcctca atagatttgt aaaataggtt tcgattagtt tcaaacaagg
     2041 gttgtttttc cgaaccgatg gctggactat ctaatggatt ttcgctcaac gccacaaaac
     2101 ttgccaaatc ttgtagcagc aatctagctt tgtcgatatt cgtttgtgtt ttgttttgta
     2161 ataaaggttc gacgtcgttc aaaatattat gcgcttttgt atttctttca tcactgtcgt
     2221 tagtgtacaa ttgactcgac gtaaacacgt taaataacgc ttggacatat ttaacatcgg
     2281 gcgtgttagc tttattaggc cgattatcgt cgtcgtccca accctcgtcg ttagaagttg
     2341 cttccgaaga cgattttgcc atagccacac gacgcctatt aattgtgtcg gctaacacgt
     2401 ccgcgatcaa atttgtagtt gagctttttg ggcctcatgg gccttccgct cactgcccgc
     2461 tttccagtcg ggaaacctgt cgtgccagct gcattaacat ggtcatagct gtttccttgc
     2521 gtattgggcg ctctccgctt cctcgctcac tgactcgctg cgctcggtcg ttcgggtaaa
     2581 gcctggggtg cctaatgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2641 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2701 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2761 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2821 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     2881 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     2941 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3001 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3061 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3121 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3181 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3241 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3301 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3361 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3421 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     3481 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     3541 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     3601 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     3661 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     3721 attgctgcag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     3781 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     3841 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     3901 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     3961 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     4021 gcgtcaacac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     4081 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4141 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4201 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4261 tgaatactca ttactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4321 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     4381 atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta
     4441 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     4501 aaatcaaaag aatagaccga gatagggttg agtggccgct acagggcgct cccattcgcc
     4561 attcaggctg cgcaactgtt gggaagggcg tttcggtgcg ggcctcttcg ctattacgcc
     4621 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     4681 agtcacgacg ttgtaaaacg acggccagtg agcgcgacgt aatacgactc actatagggc
     4741 gaattggcgg aaggccgtca aggccgtgac gttaaaacta ttaagccatc caatcgaccg
     4801 ttagtcgaat caggaccgct ggtgcgagaa gccgcgaagt atggcgaatg catcgtataa
     4861 cgtgtggagt ccgctcatta gagcgtcatg tttagacaag aaagctacat atttaattga
     4921 tcccgatgat tttattgata aattgaccct aactccatac acggtattct acaatggcgg
     4981 ggttttggtc aaaatttccg gactgcgatt gtacatgctg ttaacggctc cgcccactat
     5041 taatgaaatt aaaaattcca attttaaaaa acgcagcaag agaaacattt gtatgaaaga
     5101 atgcgtagaa ggaaagaaaa atgtcgtgga catgctgaac aacaagatta atatgcctcc
     5161 gtgtataaaa aaaatattga acgatttgaa agaaaacaat gtaccgcgcg gcggtatgta
     5221 caggaagagg tttatactaa actgttacat tgcaaacgtg gtttcgtgtg ccaagtgtga
     5281 aaaccgatgt ttaatcaagg ctctgacgca tttctacaac cacgactcca agtgtgtggg
     5341 tgaagtcatg catcttttaa tcaaatccca agatgtgtat aaaccaccaa actgccaaaa
     5401 aatgaaaact gtggacaagc tctgtccgtt tgctggcaac tgcaagggtc tcaatcctat
     5461 ttgtaattat tgaataataa aacaattata aatgctaaat ttgtttttta ttaacgatac
     5521 aaaccaaacg caacaagaac atttgtagta ttatctataa ttgaaaacgc gtagttataa
     5581 tcgctgaggt aatatttaaa atcattttca aatgattcac agttaatttg cgacaatata
     5641 attttatttt cacataaact agacgccttg tcgtcttctt cttcgtattc cttctctttt
     5701 tcatttttct cctcataaaa attaacatag ttattatcgt atccatatat gtatctatcg
     5761 tatagagtaa attttttgtt gtcataaata tatatgtctt ttttaatggg gtgtatagta
     5821 ccgctgcgca tagtttttct gtaatttaca acagtgctat tttctggtag ttcttcggag
     5881 tgtgttgctt taattattaa atttatataa tcaatgaatt tgggatcgtc ggttttgtac
     5941 aatatgttgc cggcatagta cgcagcttct tctagttcaa ttacaccatt ttttagcagc
     6001 accggattaa cataactttc caaaatgttg tacgaaccgt taaacaaaaa cagttcacct
     6061 cccttttcta tactattgtc tgcgagcagt tgtttgttgt taaaaataac agccattgta
     6121 atgagacgca caaactaata tcaca
//
