LOCUS       sequence-372986-        7690 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses PB1 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 7690)
  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 7690)
  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-PB1-B/BRISBANE/60/2008
FEATURES             Location/Qualifiers
     source          1..7690
                     /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             3215..3280
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        complement(3315..3333)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3316..3333)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3353..3370)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3359..3583
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3629..4057
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3716..3735)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3926..3947
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4066..4086)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4110..4320
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        4132..4328
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4179..4314
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4241..4260
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4395..5189
                     /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(4449..4468)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5059..5078
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5363..5484
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5400..5419)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5454..5473
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5533..5549)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5533..5549)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5546..5568)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5557..5573
                     /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(5581..5587,5588..5605,5606..5611))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5626..5647
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5764..5781)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5935..6523)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6015..6034)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6694..7485,7486..7554))
                     /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     7317..7336
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7555..7659)
                     /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 atataaatcc
      961 ttattttctc ttcatagatg tgcccgtaca ggcagcaatt tcaacaacat tcccatacac
     1021 tggtgttccc ccttattccc atggaacagg aacaggctac acaatagaca ccgtgatcag
     1081 aacgcatgag tactcaaaca aggggaaaca gtacatttct gatgttacag gatgcacaat
     1141 ggtagatcca acaaatggac cattacccga agataatgag ccgagtgcct atgcgcaatt
     1201 agattgcgtt ttagaggctt tggatagaat ggatgaagaa cacccaggtc tttttcaagc
     1261 agcctcacag aatgctatgg aggccctaat ggtcacaact gtagacaaat taacccaggg
     1321 gagacagact tttgattgga cagtatgcag aaaccaacct gctgcaacgg cactgaacac
     1381 aacaataacc tcttttaggt tgaatgattt aaatggagcc gacaaaggtg gattaatacc
     1441 tttttgccag gatatcattg attcattaga ccgacctgaa atgactttct tctcagtaaa
     1501 gaatataaag aaaaaattgc ctgccaaaaa cagaaagggt ttcctcataa agaggatacc
     1561 aatgaaggta aaagacaaaa taaccaaagt ggaatacatc aaaagagcat tatcattaaa
     1621 cacaatgaca aaagacgctg aaagaggcaa actgaaaaga agagcgattg ccactgctgg
     1681 aatacaaatc agagggtttg tattagtagt tgaaaacttg gctaaaaata tatgtgaaaa
     1741 tctagaacaa agtggtttac cagtaggtgg aaacgagaag aaagccaaac tgtcaaacgc
     1801 agtggccaaa atgctcagta actgcccacc aggagggatt agcatgacag taacaggaga
     1861 caatacaaaa tggaatgaat gtttaaaccc aagaatcttt ttggctatga ctgaaagaat
     1921 aaccagagac agcccagttt ggttcaggga tttttgtagt atagcaccgg tcctgttctc
     1981 caataagata gcaagattgg ggaaagggtt tatgataaca agcaaaacaa aaagactgaa
     2041 ggctcaaata ccttgtcctg atctgtttag tataccgtta gaaagatata atgaagaaac
     2101 aagggcaaaa ttgaaaaagc taaaaccatt cttcaatgaa gaaggaactg catctttgtc
     2161 gcctgggatg atgatgggaa tgtttaatat gctatctacc gtgttgggag tagctgcact
     2221 aggtatcaag aacattggaa acaaagaata cttatgggat ggactgcaat cttctgatga
     2281 ttttgctctg tttgttaatg caaaggatga agaaacatgt atggaaggaa taaacgactt
     2341 ttaccgaaca tgtaaattat tgggagtaaa catgagcaaa aagaaaagtt actgtaatga
     2401 gactggaatg tttgaattta caagcatgtt ctacagagat ggatttgtat ctaattttgc
     2461 aatggaactc ccttcgtttg gggttgctgg agtaaatgaa tcagcagata tggcaatagg
     2521 aatgacaata ataaagaaca acatgatcaa caatggaatg ggtccggcaa cagcacaaac
     2581 agccatacag ttattcatag ctgattatag atacacctac aaatgccaca ggggagattc
     2641 caaagtagaa ggaaagagaa tgaaaatcat aaaggagtta tgggaaaaca ctaaaggaag
     2701 agatggtcta ttagtagcag atggtgggcc caacatttac aatttgagaa acctgcatat
     2761 cccagaaata gtattaaagt ataatctaat ggaccctgaa tacaaagggc ggttacttca
     2821 tcctcaaaat ccctttgtgg gacatttgtc tattgagggc atcaaagagg cagacataac
     2881 cccagcacat ggtccagtaa agaaaatgga ctacgatgcg gtgtctggaa ctcatagttg
     2941 gagaaccaaa agaaacagat ctatactaaa cactgatcag aggaacatga ttcttgagga
     3001 acaatgctac gctaaatgtt gcaacctatt tgaggcctgt tttaacagtg catcatacag
     3061 gaagccagtg ggtcaacata gcatgcttga ggctatggcc cacagattaa gaatggatgc
     3121 acgattagat tatgaatcag ggagaatgtc aaaggatgat tttgagaaag caatggctca
     3181 ccttggtgag attgggtaca tataagcggc cgccgactac aaagaccatg acggtgatta
     3241 taaagatcat gacatcgact acaaggatga cgatgacaag taggccgctc gagcatgcat
     3301 ctagagggcc ctattctata gtgtcaccta aatgctagag ctcgctgatc agcctcgact
     3361 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     3421 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     3481 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     3541 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     3601 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     3661 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3721 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3781 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3841 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3901 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3961 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     4021 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     4081 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4141 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4201 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4261 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4321 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     4381 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4441 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4501 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4561 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4621 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4681 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4741 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     4801 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4861 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4921 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     4981 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     5041 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     5101 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     5161 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     5221 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5281 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5341 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     5401 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5461 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     5521 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5581 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5641 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5701 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5761 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5821 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5881 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5941 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6001 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6061 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6121 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6181 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6241 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6301 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6361 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6421 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6481 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6541 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6601 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6661 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6721 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6781 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6841 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6901 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6961 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     7021 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     7081 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     7141 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     7201 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     7261 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     7321 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7381 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7441 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7501 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7561 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7621 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7681 acctgacgtc
//
