LOCUS       sequence-373441-        7056 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses nucleoprotein of Influenza B/Brisbane/60/2008 virus in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7056)
  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 7056)
  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-NP-B/BRISBANE/60/2008
FEATURES             Location/Qualifiers
     source          1..7056
                     /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             2371..2382
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             2599..2640
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     promoter        complement(2681..2699)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2682..2699)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2719..2736)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2725..2949
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2995..3423
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3082..3101)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3292..3313
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3432..3452)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3476..3686
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3498..3694
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3545..3680
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3607..3626
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3761..4555
                     /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(3815..3834)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4425..4444
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4729..4850
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4766..4785)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4820..4839
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4899..4915)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4899..4915)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4912..4934)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4923..4939
                     /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(4947..4953,4954..4971,4972..4977))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4992..5013
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5130..5147)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5301..5889)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5381..5400)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6060..6851,6852..6920))
                     /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     6683..6702
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6921..7025)
                     /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 gccaccatgt ccaacatgga tattgacggt ataaacactg ggacaattga caaaacaccg
      961 gaagaaataa cttctggaac cagtgggaca accagaccaa tcattagacc agcaaccctt
     1021 gccccaccaa gcaacaaacg aacccgtaac ccatccccgg aaagagcaac cacaagcagt
     1081 gaagatgatg tcggaaggaa aacccaaaag aaacagaccc cgacagagat aaagaagagc
     1141 gtctacaaca tggtggtgaa actgggcgaa ttctataacc agatgatggt caaagctgga
     1201 ctcaatgatg acatggagag aaatctaatc caaaatgcgc atgccgtgga aagaattcta
     1261 ttggctgcca ctgatgacaa gaaaaccgag ttccagaaga aaaagaatgc cagagatgtc
     1321 aaagaaggga aagaagaaat agatcacaac aaaacaggag gcacctttta caagatggta
     1381 agagatgata aaaccatcta cttcagccct ataagaatta cctttttaaa agaagaggtg
     1441 aaaacaatgt acaaaaccac catggggagt gatggcttca gtggactaaa tcacataatg
     1501 attgggcatt cacagatgaa tgatgtctgt ttccaaagat caaaggcact aaaaagagtt
     1561 ggacttgatc cttcattaat cagtaccttt gcgggaagca cagtccccag aagatcaggt
     1621 gcgactggtg ttgcaatcaa aggaggtgga accttagtgg ctgaagccat tcgatttata
     1681 ggaagagcaa tggcagacag agggctattg agagacatca aagccaagac tgcctatgaa
     1741 aagattcttc tgaatctaaa aaacaaatgc tctgcgcccc aacaaaaggc tctagttgat
     1801 caagtgatcg gaagcagaaa tccggggatt gcagacattg aagatctaac cctgcttgct
     1861 cgtagtatgg tcgttgttag gccctctgtg gcaagcaaag tggtgcttcc cataagcatt
     1921 tacgccaaaa tacctcaact agggttcaat gttgaagagt actctatggt tgggtacgaa
     1981 gccatggctc tttacaatat ggcaacacct gtgtccatat taagaatggg agatgatgca
     2041 aaagataaat cgcaattatt cttcatgtct tgcttcggag ctgcctatga agacctgaga
     2101 gttttgtctg cattaacagg cacagaattc aagcctagat cagcattaaa atgcaagggt
     2161 ttccatgttc cagcaaagga acaggtagaa ggaatgggag cagctctgat gtccatcaag
     2221 ctccagtttt gggctccgat gaccagatct ggggggaacg aagtaggtgg agacggaggg
     2281 tctggccaaa taagctgcag cccagtgttt gcagtggaaa gacctattgc tctaagcaag
     2341 caagctgtaa gaagaatgct gtcaatgaat attgagggac gtgatgcaga tgtcaaagga
     2401 aatctactca agatgatgaa tgactcaatg gctaagaaaa ccagtggaaa tgctttcatt
     2461 gggaagaaaa tgtttcaaat atcagacaaa aacaaaacca atcccattga aattccaatt
     2521 aagcagacca tccccaattt cttctttggg agggacacag cagaggatta tgatgacctc
     2581 gattattaag gatccggagg taagcctatc cctaaccctc tcctcggtct cgattctacg
     2641 tagtaagcgg ccgctcgagc atgcatctag agggccctat tctatagtgt cacctaaatg
     2701 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     2761 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     2821 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     2881 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     2941 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     3001 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3061 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3121 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3181 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     3241 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3301 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     3361 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3421 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     3481 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3541 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3601 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     3661 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     3721 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     3781 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3841 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     3901 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     3961 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     4021 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     4081 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     4141 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     4201 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     4261 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     4321 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     4381 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     4441 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     4501 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     4561 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     4621 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4681 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     4741 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4801 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     4861 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     4921 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4981 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     5041 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     5101 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     5161 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5221 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5281 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5341 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5401 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5461 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5521 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5581 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5641 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5701 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5761 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5821 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5881 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5941 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     6001 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6061 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6121 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6181 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6241 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6301 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6361 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6421 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6481 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6541 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6601 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6661 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6721 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6781 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6841 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6901 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6961 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     7021 ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
