LOCUS       sequence-232243-        6123 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shuttle vector for baculovirus production, using FlashBac bacmid; 
            for recombinant protein with C-terminal mKate2, and C-terminal HIS6 
            tag, cleavable with 3C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6123)
  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 6123)
  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: pOCC202
FEATURES             Location/Qualifiers
     source          1..6123
                     /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             624..1319
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     CDS             1320..1343
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1353..1370
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1512..1646
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1549..1568)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1603..1622
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     3'UTR           1719..2091
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     primer_bind     complement(2443..2460)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2553..3141)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2633..2652)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3312..4103,4104..4172))
                     /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     3935..3954
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4173..4277)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4578..4600
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4592..4609
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4593..4609
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4620..4639
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4620..4638
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5249..5373
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5411..6016)
                     /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 ccgggatccg
      601 ctggctccgc tgctggttct ggcatggtga gcgagctgat taaggagaac atgcacatga
      661 agctgtacat ggagggcacc gtgaacaacc accacttcaa gtgcacatcc gagggcgaag
      721 gcaagcccta cgagggcacc cagaccatga gaatcaaggc ggtcgagggc ggccctctcc
      781 ccttcgcctt cgacatcctg gctaccagct tcatgtacgg cagcaaaacc ttcatcaacc
      841 acacccaggg catccccgac ttctttaagc agtccttccc cgagggcttc acatgggaga
      901 gagtcaccac atacgaagac gggggcgtgc tgaccgctac ccaggacacc agcctccagg
      961 acggctgcct catctacaac gtcaagatca gaggggtgaa cttcccatcc aacggccctg
     1021 tgatgcagaa gaaaacactc ggctgggagg cctccaccga gaccctgtac cccgctgacg
     1081 gcggcctgga aggcagagcc gacatggccc tgaagctcgt gggcgggggc cacctgatct
     1141 gcaacttgaa gaccacatac agatccaaga aacccgctaa gaacctcaag atgcccggcg
     1201 tctactatgt ggacagaaga ctggaaagaa tcaaggaggc cgacaaagag acctacgtcg
     1261 agcagcacga ggtggctgtg gccagatact gcgacctccc tagcaaactg gggcacagac
     1321 tggaagttct gttccagggg ccgggcagca gccatcacca tcaccatcac agtggataat
     1381 agaagcttgt cgagaagtac tagaggatca taatcagcca taccacattt gtagaggttt
     1441 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     1501 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     1561 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     1621 tcaatgtatc ttatcatgtc tggatctgat cactgcttga gcctaggaga tccgaaccag
     1681 ataagtgaaa gcctgcatcc gcagatctta attaataaaa cacgatacat tgttattagt
     1741 acatttatta agcgctagat tctgtgcgtt gttgatttac agacaattgt tgtacgtatt
     1801 ttaataattc attaaattta taatctttag ggtggtatgt tagagcgaaa atcaaatgat
     1861 tttcagcgtc tttatatctg aatttaaata ttaaatcctc aatagatttg taaaataggt
     1921 ttcgattagt ttcaaacaag ggttgttttt ccgaaccgat ggctggacta tctaatggat
     1981 tttcgctcaa cgccacaaaa cttgccaaat cttgtagcag caatctagct ttgtcgatat
     2041 tcgtttgtgt tttgttttgt aataaaggtt cgacgtcgtt caaaatatta tgcgcttttg
     2101 tatttctttc atcactgtcg ttagtgtaca attgactcga cgtaaacacg ttaaataacg
     2161 cttggacata tttaacatcg ggcgtgttag ctttattagg ccgattatcg tcgtcgtccc
     2221 aaccctcgtc gttagaagtt gcttccgaag acgattttgc catagccaca cgacgcctat
     2281 taattgtgtc ggctaacacg tccgcgatca aatttgtagt tgagcttttt gggcctcatg
     2341 ggccttccgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaaca
     2401 tggtcatagc tgtttccttg cgtattgggc gctctccgct tcctcgctca ctgactcgct
     2461 gcgctcggtc gttcgggtaa agcctggggt gcctaatgag caaaaggcca gcaaaaggcc
     2521 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2581 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2641 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2701 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2761 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2821 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     2881 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     2941 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     3001 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3061 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3121 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3181 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3241 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3301 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3361 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     3421 ccatctggcc ccagtgctgc aatgataccg cgagaaccac gctcaccggc tccagattta
     3481 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3541 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     3601 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     3661 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     3721 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     3781 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     3841 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     3901 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     3961 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4021 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4081 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4141 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     4201 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     4261 caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata
     4321 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     4381 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtggccgct
     4441 acagggcgct cccattcgcc attcaggctg cgcaactgtt gggaagggcg tttcggtgcg
     4501 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     4561 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg agcgcgacgt
     4621 aatacgactc actatagggc gaattggcgg aaggccgtca aggccgtgac gttaaaacta
     4681 ttaagccatc caatcgaccg ttagtcgaat caggaccgct ggtgcgagaa gccgcgaagt
     4741 atggcgaatg catcgtataa cgtgtggagt ccgctcatta gagcgtcatg tttagacaag
     4801 aaagctacat atttaattga tcccgatgat tttattgata aattgaccct aactccatac
     4861 acggtattct acaatggcgg ggttttggtc aaaatttccg gactgcgatt gtacatgctg
     4921 ttaacggctc cgcccactat taatgaaatt aaaaattcca attttaaaaa acgcagcaag
     4981 agaaacattt gtatgaaaga atgcgtagaa ggaaagaaaa atgtcgtgga catgctgaac
     5041 aacaagatta atatgcctcc gtgtataaaa aaaatattga acgatttgaa agaaaacaat
     5101 gtaccgcgcg gcggtatgta caggaagagg tttatactaa actgttacat tgcaaacgtg
     5161 gtttcgtgtg ccaagtgtga aaaccgatgt ttaatcaagg ctctgacgca tttctacaac
     5221 cacgactcca agtgtgtggg tgaagtcatg catcttttaa tcaaatccca agatgtgtat
     5281 aaaccaccaa actgccaaaa aatgaaaact gtggacaagc tctgtccgtt tgctggcaac
     5341 tgcaagggtc tcaatcctat ttgtaattat tgaataataa aacaattata aatgctaaat
     5401 ttgtttttta ttaacgatac aaaccaaacg caacaagaac atttgtagta ttatctataa
     5461 ttgaaaacgc gtagttataa tcgctgaggt aatatttaaa atcattttca aatgattcac
     5521 agttaatttg cgacaatata attttatttt cacataaact agacgccttg tcgtcttctt
     5581 cttcgtattc cttctctttt tcatttttct cctcataaaa attaacatag ttattatcgt
     5641 atccatatat gtatctatcg tatagagtaa attttttgtt gtcataaata tatatgtctt
     5701 ttttaatggg gtgtatagta ccgctgcgca tagtttttct gtaatttaca acagtgctat
     5761 tttctggtag ttcttcggag tgtgttgctt taattattaa atttatataa tcaatgaatt
     5821 tgggatcgtc ggttttgtac aatatgttgc cggcatagta cgcagcttct tctagttcaa
     5881 ttacaccatt ttttagcagc accggattaa cataactttc caaaatgttg tacgaaccgt
     5941 taaacaaaaa cagttcacct cccttttcta tactattgtc tgcgagcagt tgtttgttgt
     6001 taaaaataac agccattgta atgagacgca caaactaata tcacaagctg ccaccatttt
     6061 gttcgcccag gactctagct atagttctag tggttggcta cgtatactcc ggaatattaa
     6121 tag
//
