LOCUS       sequence-373428-        7744 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses PB2 subunit of Influenza B/Brisbane/60/2008 virus 
            polymerase in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7744)
  AUTHORS   Kedia N, Banerjee S, Mondal A
  TITLE     A Comprehensive Roadmap Towards the Generation of an Influenza B 
            Reporter Assay Using a Single DNA Polymerase-Based Cloning of the 
            Reporter RNA Construct.
  JOURNAL   Front Microbiol. 2022 May 25;13:868367. doi: 
            10.3389/fmicb.2022.868367. eCollection 2022.
  PUBMED    35694292
REFERENCE   2  (bases 1 to 7744)
  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: "Front 
            Microbiol."; date: "2022-05-25"; volume: "13:868367. doi"; pages: " 
            10.3389/fmicb.2022.868367. eCollection 2022"
COMMENT     Sequence Label: pcDNA3-PB2-B/BRISBANE/60/2008
FEATURES             Location/Qualifiers
     source          1..7744
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3269..3334
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        complement(3369..3387)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3370..3387)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3407..3424)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3413..3637
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3683..4111
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3770..3789)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3980..4001
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4120..4140)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4164..4374
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        4186..4382
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4233..4368
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4295..4314
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4449..5243
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4503..4522)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5113..5132
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5417..5538
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5454..5473)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5508..5527
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5587..5603)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5587..5603)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5600..5622)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5611..5627
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5635..5641,5642..5659,5660..5665))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5680..5701
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5818..5835)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5989..6577)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6069..6088)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6748..7539,7540..7608))
                     /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     7371..7390
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7609..7713)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttcaccatga cattggccaa
      961 aattgaattg ttaaaacaac tgctaaggga caatgaagcc aaaacagttt tgaagcaaac
     1021 aacggtagac caatataaca taataagaaa attcaataca tcaaggattg aaaagaatcc
     1081 ttcactaagg atgaagtggg ccatgtgttc taattttccc ttggctctaa ccaagggcga
     1141 tatggcaaac agaatcccct tggaatacaa agggatacaa cttaaaacaa atgctgaaga
     1201 cataggaacc aaaggccaaa tgtgctcaat agcagcagtt acttggtgga atacatatgg
     1261 accaatagga gatactgaag gtttcgaaag ggtctacgaa agcttttttc tcagaaaaat
     1321 gagacttgac aacgccactt ggggccgaat aacttttggc ccagttgaaa gagtgagaaa
     1381 aagggtactg ctaaaccctc tcaccaagga aatgcctccg gatgaggcga gcaatgtgat
     1441 aatggaaata ttgttcccta aagaagcagg aataccaaga gaatccactt ggatacatag
     1501 ggaactgata aaagaaaaaa gagaaaaatt gaaaggaaca atgataactc caatcgtact
     1561 ggcatacatg cttgaaagag aactggttgc tcgaagaaga ttcttgccag tggcaggagc
     1621 aacatcagct gagttcatag aaatgctaca ctgcttacaa ggtgaaaatt ggagacaaat
     1681 atatcaccca ggagggaata aattaactga gtccaggtct caatcaatga tagtagcttg
     1741 tagaaaaata atcagaagat caatagtcgc ttcaaaccca ctggagctag ctgtagaaat
     1801 tgcaaacaag actgtgatag atactgaacc tttaaagtca tgtctggcag ccatagacgg
     1861 aggtgatgta gcttgtgaca taataagagc tgcattagga ctaaagatca gacaaagaca
     1921 aagatttgga cggcttgagc taaaaagaat atcaggaaga ggattcaaaa atgatgaaga
     1981 aatattaata gggaacggaa caatacagaa gattggaata tgggacgggg aagaggagtt
     2041 ccatgtaaga tgtggtgaat gcaggggaat attaaaaaag agtaaaatga aactggaaaa
     2101 actactgata aattcagcca aaaaggagga tatgagagat ttaataatct tatgcatggt
     2161 attttctcaa gacactagga tgttccaagg agtgagagga gaaataaatt ttcttaatcg
     2221 agcaggccaa cttttatctc caatgtacca actccaacga tattttttga atagaagcaa
     2281 cgaccttttt gatcaatggg ggtatgagga atcacccaaa gcaagtgaac tacatgggat
     2341 aaatgaatca atgaatgcat ctgactatac attgaaaggg attgtagtga caagaaatgt
     2401 aattgacgac tttagctcta ttgaaacaga aaaagtatcc ataacaaaaa atcttagttt
     2461 aataaaaagg actggggaag tcataatggg agctaatgac gtgagtgaat tagaatcaca
     2521 agcacagctg atgataacat atgatacacc taaaatgtgg gaaatgggaa caaccaaaga
     2581 actggtgcaa aacacttatc aatgggtgct aaaaaacttg gtgacactga aggctcagtt
     2641 tcttctagga aaagaggaca tgttccaatg ggatgcattt gaagcatttg agagcataat
     2701 tcctcagaag atggctggtc agtacagtgg atttgcaaga gcagtgctca aacaaatgag
     2761 agaccaggag gttatgaaaa ctgaccagtt cataaagttg ttgccttttt gtttctcacc
     2821 accaaaatta aggagcaatg gggagcctta tcaattctta aaacttgtgt tgaaaggagg
     2881 aggggaaaat ttcatcgaag taaggaaagg gtcccctcta ttttcctata atccacaaac
     2941 agaagtccta actatatgcg gcagaatgat gtcattaaaa gggaaaattg aagatgaaga
     3001 aaggaataga tcaatgggta atgcagtatt agcaggcttt ctcgttagtg gcaagtatga
     3061 cccagatctt ggagatttca aaactattga agaacttgaa aagctgaaac cgggggaaaa
     3121 ggcaaacatc ttactttatc aaggaaaacc agttaaagta gttaaaagga aaaggtatag
     3181 tgctttgtcc aatgacattt cacaaggaat taagagacaa agaatgacag ttgagtctat
     3241 ggggtgggcc ttgagctaag cggccgccga ctacaaagac catgacggtg attataaaga
     3301 tcatgacatc gactacaagg atgacgatga caagtaggcc gctcgagcat gcatctagag
     3361 ggccctattc tatagtgtca cctaaatgct agagctcgct gatcagcctc gactgtgcct
     3421 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     3481 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     3541 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     3601 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     3661 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3721 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3781 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3841 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3901 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3961 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     4021 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     4081 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     4141 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4201 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     4261 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4321 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4381 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     4441 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4501 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4561 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4621 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4681 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4741 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4801 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4861 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4921 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     4981 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     5041 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     5101 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     5161 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     5221 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     5281 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     5341 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     5401 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5461 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5521 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     5581 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     5641 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     5701 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     5761 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     5821 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     5881 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5941 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     6001 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     6061 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     6121 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     6181 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     6241 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     6301 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6361 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6421 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6481 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6541 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6601 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6661 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6721 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6781 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6841 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6901 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6961 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     7021 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     7081 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     7141 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     7201 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     7261 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     7321 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     7381 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7441 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7501 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7561 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7621 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7681 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7741 cgtc
//
