LOCUS       sequence-232355-        6153 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with C-terminal monomeric GFP, cleavable 
            with TEV, and C-terminal HIS6 tag, cleavable with 3C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6153)
  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 6153)
  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: pOCC70
FEATURES             Location/Qualifiers
     source          1..6153
                     /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             612..632
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(633..635,636..638,639..1349)
                     /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(678..699)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(939..958)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1286..1307
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1350..1373
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1383..1400
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1542..1676
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1579..1598)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1633..1652
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1749..2121
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(2473..2490)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2583..3171)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2663..2682)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3342..4133,4134..4202))
                     /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     3965..3984
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4203..4307)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4608..4630
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4622..4639
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4623..4639
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4650..4669
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4650..4668
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5279..5403
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5441..6046)
                     /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 ccgggatcca
      601 gcagcggcag agaaaacttg tatttccagg gcatggtgag caagggcgag gagctgttca
      661 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
      721 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
      781 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
      841 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
      901 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
      961 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     1021 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     1081 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     1141 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     1201 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc aaactgagca
     1261 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     1321 tcactctcgg catggacgag ctgtacaagc tggaagttct gttccagggg ccgggcagca
     1381 gccatcacca tcaccatcac agtggataat agaagcttgt cgagaagtac tagaggatca
     1441 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     1501 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     1561 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     1621 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatctgat
     1681 cactgcttga gcctaggaga tccgaaccag ataagtgaaa gcctgcatcc gcagatctta
     1741 attaataaaa cacgatacat tgttattagt acatttatta agcgctagat tctgtgcgtt
     1801 gttgatttac agacaattgt tgtacgtatt ttaataattc attaaattta taatctttag
     1861 ggtggtatgt tagagcgaaa atcaaatgat tttcagcgtc tttatatctg aatttaaata
     1921 ttaaatcctc aatagatttg taaaataggt ttcgattagt ttcaaacaag ggttgttttt
     1981 ccgaaccgat ggctggacta tctaatggat tttcgctcaa cgccacaaaa cttgccaaat
     2041 cttgtagcag caatctagct ttgtcgatat tcgtttgtgt tttgttttgt aataaaggtt
     2101 cgacgtcgtt caaaatatta tgcgcttttg tatttctttc atcactgtcg ttagtgtaca
     2161 attgactcga cgtaaacacg ttaaataacg cttggacata tttaacatcg ggcgtgttag
     2221 ctttattagg ccgattatcg tcgtcgtccc aaccctcgtc gttagaagtt gcttccgaag
     2281 acgattttgc catagccaca cgacgcctat taattgtgtc ggctaacacg tccgcgatca
     2341 aatttgtagt tgagcttttt gggcctcatg ggccttccgc tcactgcccg ctttccagtc
     2401 gggaaacctg tcgtgccagc tgcattaaca tggtcatagc tgtttccttg cgtattgggc
     2461 gctctccgct tcctcgctca ctgactcgct gcgctcggtc gttcgggtaa agcctggggt
     2521 gcctaatgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2581 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2641 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2701 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2761 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     2821 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     2881 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     2941 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     3001 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     3061 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3121 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     3181 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     3241 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     3301 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     3361 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     3421 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     3481 cgagaaccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     3541 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     3601 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     3661 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     3721 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     3781 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     3841 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     3901 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     3961 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     4021 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     4081 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     4141 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     4201 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     4261 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     4321 aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     4381 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     4441 aatagaccga gatagggttg agtggccgct acagggcgct cccattcgcc attcaggctg
     4501 cgcaactgtt gggaagggcg tttcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     4561 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     4621 ttgtaaaacg acggccagtg agcgcgacgt aatacgactc actatagggc gaattggcgg
     4681 aaggccgtca aggccgtgac gttaaaacta ttaagccatc caatcgaccg ttagtcgaat
     4741 caggaccgct ggtgcgagaa gccgcgaagt atggcgaatg catcgtataa cgtgtggagt
     4801 ccgctcatta gagcgtcatg tttagacaag aaagctacat atttaattga tcccgatgat
     4861 tttattgata aattgaccct aactccatac acggtattct acaatggcgg ggttttggtc
     4921 aaaatttccg gactgcgatt gtacatgctg ttaacggctc cgcccactat taatgaaatt
     4981 aaaaattcca attttaaaaa acgcagcaag agaaacattt gtatgaaaga atgcgtagaa
     5041 ggaaagaaaa atgtcgtgga catgctgaac aacaagatta atatgcctcc gtgtataaaa
     5101 aaaatattga acgatttgaa agaaaacaat gtaccgcgcg gcggtatgta caggaagagg
     5161 tttatactaa actgttacat tgcaaacgtg gtttcgtgtg ccaagtgtga aaaccgatgt
     5221 ttaatcaagg ctctgacgca tttctacaac cacgactcca agtgtgtggg tgaagtcatg
     5281 catcttttaa tcaaatccca agatgtgtat aaaccaccaa actgccaaaa aatgaaaact
     5341 gtggacaagc tctgtccgtt tgctggcaac tgcaagggtc tcaatcctat ttgtaattat
     5401 tgaataataa aacaattata aatgctaaat ttgtttttta ttaacgatac aaaccaaacg
     5461 caacaagaac atttgtagta ttatctataa ttgaaaacgc gtagttataa tcgctgaggt
     5521 aatatttaaa atcattttca aatgattcac agttaatttg cgacaatata attttatttt
     5581 cacataaact agacgccttg tcgtcttctt cttcgtattc cttctctttt tcatttttct
     5641 cctcataaaa attaacatag ttattatcgt atccatatat gtatctatcg tatagagtaa
     5701 attttttgtt gtcataaata tatatgtctt ttttaatggg gtgtatagta ccgctgcgca
     5761 tagtttttct gtaatttaca acagtgctat tttctggtag ttcttcggag tgtgttgctt
     5821 taattattaa atttatataa tcaatgaatt tgggatcgtc ggttttgtac aatatgttgc
     5881 cggcatagta cgcagcttct tctagttcaa ttacaccatt ttttagcagc accggattaa
     5941 cataactttc caaaatgttg tacgaaccgt taaacaaaaa cagttcacct cccttttcta
     6001 tactattgtc tgcgagcagt tgtttgttgt taaaaataac agccattgta atgagacgca
     6061 caaactaata tcacaagctg ccaccatttt gttcgcccag gactctagct atagttctag
     6121 tggttggcta cgtatactcc ggaatattaa tag
//
