LOCUS       sequence-232312-        5457 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with N-terminal TwinStrep, cleavable with 
            3C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5457)
  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 5457)
  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: pOCC153
FEATURES             Location/Qualifiers
     source          1..5457
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      42..51
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(54..59,60..83,84..119,120..143)
                     /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"
     CDS             144..167
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             276..581
                     /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     460..479
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     polyA_signal    754..888
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(791..810)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     845..864
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           961..1333
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(1685..1702)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1795..2383)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1875..1894)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2554..3345,3346..3414))
                     /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     3177..3196
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3415..3519)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3820..3842
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3834..3851
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3835..3851
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3862..3881
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3862..3880
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4491..4615
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(4653..5258)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     promoter        5366..5457
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     5382..5399
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
ORIGIN
        1 attccggatt attcataccg tcccaccatc gggcgcggat cgccaccatg ggcagcgctt
       61 ggagccatcc gcagtttgaa aaaggtggtg gtagcggtgg tggttcaggt ggtagtgcat
      121 ggtcacaccc tcagtttgag aaactggaag ttctgtttca gggtcctgcg gccgcaagcg
      181 ctatggccac aacctgttac attgcacaag ataaaaatat atcatcacga acagtaaaac
      241 tgtctgctta cataaacagt aatacaaggg gtgttatgca gtttaaggtt tacacctata
      301 aaagagagag ccgttatcgt ctgtttgtgg atgtacagag tgatattatt gacacgcccg
      361 ggcgacggat ggtgatcccc ctggccagtg cccgtctgct gtcagataaa gtctcccgtg
      421 aactttaccc ggtggtgcat atcggggatg aaagctggcg catgatgacc accgatatgg
      481 ccagtgtgcc ggtctccgtt atcggggaag aagtggctga tctcagccac cgcgaaaatg
      541 acatcaaaaa cgccattaac ctgatgttct ggggaatata aatgtcaggc tcccttatac
      601 acagccagtg gcgcgccgta atagaagctt gtcgagaagt actagaggat cataatcagc
      661 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
      721 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
      781 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
      841 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatctg atcactgctt
      901 gagcctagga gatccgaacc agataagtga aagcctgcat ccgcagatct taattaataa
      961 aacacgatac attgttatta gtacatttat taagcgctag attctgtgcg ttgttgattt
     1021 acagacaatt gttgtacgta ttttaataat tcattaaatt tataatcttt agggtggtat
     1081 gttagagcga aaatcaaatg attttcagcg tctttatatc tgaatttaaa tattaaatcc
     1141 tcaatagatt tgtaaaatag gtttcgatta gtttcaaaca agggttgttt ttccgaaccg
     1201 atggctggac tatctaatgg attttcgctc aacgccacaa aacttgccaa atcttgtagc
     1261 agcaatctag ctttgtcgat attcgtttgt gttttgtttt gtaataaagg ttcgacgtcg
     1321 ttcaaaatat tatgcgcttt tgtatttctt tcatcactgt cgttagtgta caattgactc
     1381 gacgtaaaca cgttaaataa cgcttggaca tatttaacat cgggcgtgtt agctttatta
     1441 ggccgattat cgtcgtcgtc ccaaccctcg tcgttagaag ttgcttccga agacgatttt
     1501 gccatagcca cacgacgcct attaattgtg tcggctaaca cgtccgcgat caaatttgta
     1561 gttgagcttt ttgggcctca tgggccttcc gctcactgcc cgctttccag tcgggaaacc
     1621 tgtcgtgcca gctgcattaa catggtcata gctgtttcct tgcgtattgg gcgctctccg
     1681 cttcctcgct cactgactcg ctgcgctcgg tcgttcgggt aaagcctggg gtgcctaatg
     1741 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     1801 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     1861 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     1921 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     1981 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2041 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2101 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2161 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2221 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2281 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2341 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2401 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     2461 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     2521 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     2581 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     2641 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaacc
     2701 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     2761 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     2821 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     2881 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     2941 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     3001 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     3061 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     3121 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     3181 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     3241 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     3301 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     3361 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     3421 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     3481 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     3541 acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     3601 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     3661 gagatagggt tgagtggccg ctacagggcg ctcccattcg ccattcaggc tgcgcaactg
     3721 ttgggaaggg cgtttcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
     3781 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
     3841 cgacggccag tgagcgcgac gtaatacgac tcactatagg gcgaattggc ggaaggccgt
     3901 caaggccgtg acgttaaaac tattaagcca tccaatcgac cgttagtcga atcaggaccg
     3961 ctggtgcgag aagccgcgaa gtatggcgaa tgcatcgtat aacgtgtgga gtccgctcat
     4021 tagagcgtca tgtttagaca agaaagctac atatttaatt gatcccgatg attttattga
     4081 taaattgacc ctaactccat acacggtatt ctacaatggc ggggttttgg tcaaaatttc
     4141 cggactgcga ttgtacatgc tgttaacggc tccgcccact attaatgaaa ttaaaaattc
     4201 caattttaaa aaacgcagca agagaaacat ttgtatgaaa gaatgcgtag aaggaaagaa
     4261 aaatgtcgtg gacatgctga acaacaagat taatatgcct ccgtgtataa aaaaaatatt
     4321 gaacgatttg aaagaaaaca atgtaccgcg cggcggtatg tacaggaaga ggtttatact
     4381 aaactgttac attgcaaacg tggtttcgtg tgccaagtgt gaaaaccgat gtttaatcaa
     4441 ggctctgacg catttctaca accacgactc caagtgtgtg ggtgaagtca tgcatctttt
     4501 aatcaaatcc caagatgtgt ataaaccacc aaactgccaa aaaatgaaaa ctgtggacaa
     4561 gctctgtccg tttgctggca actgcaaggg tctcaatcct atttgtaatt attgaataat
     4621 aaaacaatta taaatgctaa atttgttttt tattaacgat acaaaccaaa cgcaacaaga
     4681 acatttgtag tattatctat aattgaaaac gcgtagttat aatcgctgag gtaatattta
     4741 aaatcatttt caaatgattc acagttaatt tgcgacaata taattttatt ttcacataaa
     4801 ctagacgcct tgtcgtcttc ttcttcgtat tccttctctt tttcattttt ctcctcataa
     4861 aaattaacat agttattatc gtatccatat atgtatctat cgtatagagt aaattttttg
     4921 ttgtcataaa tatatatgtc ttttttaatg gggtgtatag taccgctgcg catagttttt
     4981 ctgtaattta caacagtgct attttctggt agttcttcgg agtgtgttgc tttaattatt
     5041 aaatttatat aatcaatgaa tttgggatcg tcggttttgt acaatatgtt gccggcatag
     5101 tacgcagctt cttctagttc aattacacca ttttttagca gcaccggatt aacataactt
     5161 tccaaaatgt tgtacgaacc gttaaacaaa aacagttcac ctcccttttc tatactattg
     5221 tctgcgagca gttgtttgtt gttaaaaata acagccattg taatgagacg cacaaactaa
     5281 tatcacaagc tgccaccatt ttgttcgccc aggactctag ctatagttct agtggttggc
     5341 tacgtatact ccggaatatt aatagatcat ggagataatt aaaatgataa ccatctcgca
     5401 aataaataag tattttactg ttttcgtaac agttttgtaa taaaaaaacc tataaat
//
