LOCUS       sequence-224695-        5422 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with N-terminal HIS6, cleavable with 3C, and
            C-terminal sortase site (LPETGg).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5422)
  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 5422)
  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: pOCC110
FEATURES             Location/Qualifiers
     source          1..5422
                     /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"
     CDS             230..247
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             260..283
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             392..697
                     /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     576..595
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    888..1022
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(925..944)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     979..998
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1095..1467
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(1819..1836)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1929..2517)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2009..2028)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2688..3479,3480..3548))
                     /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     3311..3330
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3550..3654)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3955..3977
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3969..3986
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3970..3986
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3997..4016
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3997..4015
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4626..4750
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(4788..5393)
                     /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 ggcagcagcc atcaccatca
      241 ccatcacagc agcggcagac tggaagttct gttccagggg ccggcggccg caagcgctat
      301 ggccacaacc tgttacattg cacaagataa aaatatatca tcacgaacag taaaactgtc
      361 tgcttacata aacagtaata caaggggtgt tatgcagttt aaggtttaca cctataaaag
      421 agagagccgt tatcgtctgt ttgtggatgt acagagtgat attattgaca cgcccgggcg
      481 acggatggtg atccccctgg ccagtgcccg tctgctgtca gataaagtct cccgtgaact
      541 ttacccggtg gtgcatatcg gggatgaaag ctggcgcatg atgaccaccg atatggccag
      601 tgtgccggtc tccgttatcg gggaagaagt ggctgatctc agccaccgcg aaaatgacat
      661 caaaaacgcc attaacctga tgttctgggg aatataaatg tcaggctccc ttatacacag
      721 ccagtggcgc gccgctgccc gagaccggcg gctaatagaa gcttgtcgag aagtactaga
      781 ggatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
      841 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
      901 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
      961 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     1021 tctgatcact gcttgagcct aggagatccg aaccagataa gtgaaagcct gcatccgcag
     1081 atcttaatta ataaaacacg atacattgtt attagtacat ttattaagcg ctagattctg
     1141 tgcgttgttg atttacagac aattgttgta cgtattttaa taattcatta aatttataat
     1201 ctttagggtg gtatgttaga gcgaaaatca aatgattttc agcgtcttta tatctgaatt
     1261 taaatattaa atcctcaata gatttgtaaa ataggtttcg attagtttca aacaagggtt
     1321 gtttttccga accgatggct ggactatcta atggattttc gctcaacgcc acaaaacttg
     1381 ccaaatcttg tagcagcaat ctagctttgt cgatattcgt ttgtgttttg ttttgtaata
     1441 aaggttcgac gtcgttcaaa atattatgcg cttttgtatt tctttcatca ctgtcgttag
     1501 tgtacaattg actcgacgta aacacgttaa ataacgcttg gacatattta acatcgggcg
     1561 tgttagcttt attaggccga ttatcgtcgt cgtcccaacc ctcgtcgtta gaagttgctt
     1621 ccgaagacga ttttgccata gccacacgac gcctattaat tgtgtcggct aacacgtccg
     1681 cgatcaaatt tgtagttgag ctttttgggc ctcatgggcc ttccgctcac tgcccgcttt
     1741 ccagtcggga aacctgtcgt gccagctgca ttaacatggt catagctgtt tccttgcgta
     1801 ttgggcgctc tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc gggtaaagcc
     1861 tggggtgcct aatgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     1921 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     1981 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2041 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     2101 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     2161 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     2221 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     2281 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     2341 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     2401 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     2461 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     2521 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     2581 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     2641 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     2701 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     2761 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     2821 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     2881 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     2941 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     3001 gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     3061 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     3121 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     3181 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     3241 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     3301 tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     3361 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     3421 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     3481 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     3541 atactcatta ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     3601 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     3661 tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     3721 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     3781 tcaaaagaat agaccgagat agggttgagt ggccgctaca gggcgctccc attcgccatt
     3841 caggctgcgc aactgttggg aagggcgttt cggtgcgggc ctcttcgcta ttacgccagc
     3901 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
     3961 cacgacgttg taaaacgacg gccagtgagc gcgacgtaat acgactcact atagggcgaa
     4021 ttggcggaag gccgtcaagg ccgtgacgtt aaaactatta agccatccaa tcgaccgtta
     4081 gtcgaatcag gaccgctggt gcgagaagcc gcgaagtatg gcgaatgcat cgtataacgt
     4141 gtggagtccg ctcattagag cgtcatgttt agacaagaaa gctacatatt taattgatcc
     4201 cgatgatttt attgataaat tgaccctaac tccatacacg gtattctaca atggcggggt
     4261 tttggtcaaa atttccggac tgcgattgta catgctgtta acggctccgc ccactattaa
     4321 tgaaattaaa aattccaatt ttaaaaaacg cagcaagaga aacatttgta tgaaagaatg
     4381 cgtagaagga aagaaaaatg tcgtggacat gctgaacaac aagattaata tgcctccgtg
     4441 tataaaaaaa atattgaacg atttgaaaga aaacaatgta ccgcgcggcg gtatgtacag
     4501 gaagaggttt atactaaact gttacattgc aaacgtggtt tcgtgtgcca agtgtgaaaa
     4561 ccgatgttta atcaaggctc tgacgcattt ctacaaccac gactccaagt gtgtgggtga
     4621 agtcatgcat cttttaatca aatcccaaga tgtgtataaa ccaccaaact gccaaaaaat
     4681 gaaaactgtg gacaagctct gtccgtttgc tggcaactgc aagggtctca atcctatttg
     4741 taattattga ataataaaac aattataaat gctaaatttg ttttttatta acgatacaaa
     4801 ccaaacgcaa caagaacatt tgtagtatta tctataattg aaaacgcgta gttataatcg
     4861 ctgaggtaat atttaaaatc attttcaaat gattcacagt taatttgcga caatataatt
     4921 ttattttcac ataaactaga cgccttgtcg tcttcttctt cgtattcctt ctctttttca
     4981 tttttctcct cataaaaatt aacatagtta ttatcgtatc catatatgta tctatcgtat
     5041 agagtaaatt ttttgttgtc ataaatatat atgtcttttt taatggggtg tatagtaccg
     5101 ctgcgcatag tttttctgta atttacaaca gtgctatttt ctggtagttc ttcggagtgt
     5161 gttgctttaa ttattaaatt tatataatca atgaatttgg gatcgtcggt tttgtacaat
     5221 atgttgccgg catagtacgc agcttcttct agttcaatta caccattttt tagcagcacc
     5281 ggattaacat aactttccaa aatgttgtac gaaccgttaa acaaaaacag ttcacctccc
     5341 ttttctatac tattgtctgc gagcagttgt ttgttgttaa aaataacagc cattgtaatg
     5401 agacgcacaa actaatatca ca
//
