LOCUS       sequence-392592-        7609 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses PA 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 7609)
  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 7609)
  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-PA-B/BRISBANE/60/2008
FEATURES             Location/Qualifiers
     source          1..7609
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(49..68)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        240..619
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        620..823
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     774..794
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     868..887
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        868..886
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3139..3204
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        complement(3239..3257)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3240..3257)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3277..3294)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3283..3507
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3553..3981
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3640..3659)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3850..3871
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3990..4010)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4034..4244
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        4056..4252
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4103..4238
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4165..4184
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4319..5113
                     /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(4373..4392)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4983..5002
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5287..5408
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5324..5343)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5378..5397
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5457..5473)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5457..5473)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5470..5492)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5481..5497
                     /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(5505..5511,5512..5529,5530..5535))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5550..5571
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5688..5705)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5859..6447)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5939..5958)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6618..7409,7410..7478))
                     /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     7241..7260
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7479..7583)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc
       61 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
      121 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
      181 tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg
      241 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
      301 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      361 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      421 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      481 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      541 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      601 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      661 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      721 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      781 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga
      841 acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga cccaagcttg
      901 gtaccgagct cggatccact agtaacggcc gccagtgtgc tggaattcac catggatact
      961 tttattacaa gaaacttcca gactacaata atacaaaagg ccaaaaacac aatggcagaa
     1021 tttagtgaag atcctgaatt gcaaccagca atgctattca atatctgcgt ccatctagag
     1081 gtttgctatg taataagtga catgaatttt cttgacgaag aaggaaaagc atatacagca
     1141 ttagaaggac aagggaaaga acaaaacttg agaccacaat atgaagtaat tgagggaatg
     1201 ccaagaacca tagcatggat ggtccagaga tccttagctc aagagcatgg aatagagact
     1261 cccaagtatc tggctgattt gtttgattat aaaaccaaaa gatttataga agttggaata
     1321 acaaagggat tggctgatga ttacttttgg aaaaagaaag aaaagttggg aaatagcatg
     1381 gaactgatga tattcagcta caatcaagac tactcgttaa gtaatgaatc ctcattggat
     1441 gaggaaggga aagggagagt gctaagcaga ctcacagaac ttcaggctga attaagtctg
     1501 aaaaatttat ggcaagttct cataggagaa gaagatgttg aaaagggaat tgattttaaa
     1561 cttggacaaa caatatctag actaagggat atatctgttc cagctggttt ctccaatttt
     1621 gaaggaatga ggagctacat agacaatata gacccaaaag gagcaataga gagaaatcta
     1681 gcaaggatgt ctcccttagt atcagtcaca cctaaaaagt taacatggga ggacctaaga
     1741 ccaatagggc ctcacattta cgaccatgag ctaccagaag ttccatataa tgcctttctt
     1801 ctaatgtctg atgaactggg attggccaat atgactgagg gaaagtccaa aaaaccgaag
     1861 acattagcca aagaatgtct agaaaagtac tcaacactac gggatcaaac tgacccaata
     1921 ttaataatga aaagcgaaaa agctaacgaa aatttcctat ggaagctttg gagagactgt
     1981 gtaaatacaa taagtaatga ggaaacgagt aacgagttac agaaaaccaa ttatgccaaa
     2041 tgggccacag gggatggatt aacataccag aaaataatga aagaagtagc aatagatgac
     2101 gaaacaatgt gccaagaaga gcctaaaatc cctaacaaat gtagagtggc tgcttgggtt
     2161 caaacagaga tgaatctatt gagcactctg acaagtaaaa gagctctgga cctaccagaa
     2221 atagggccag acatagcacc cgtggagcat gtaggaagtg aaagaaggaa atactttgtt
     2281 aatgaaatca actactgtaa ggcctctaca gttatgatga agtatgtgct ttttcacact
     2341 tcattgttga atgaaagcaa tgccagcatg ggaaaataca aagtaatacc aataaccaat
     2401 agagtagtaa atgaaaaagg agaaagtttc gacatgcttt acggtctggc ggttaaagga
     2461 caatctcatc tgaggggaga tactgatgtt gtaacagttg taactttcga atttagtagt
     2521 acagatccaa gagtggactc aggaaagtgg ccaaaatata ctgtgtttag gattggctcc
     2581 ctatttgtga gtgggaggga aaaatctgtg tacttgtatt gccgagtgaa tggcacaaat
     2641 aagatccaaa tgaaatgggg aatggaagct agaagatgtt tgcttcaatc aatgcaacaa
     2701 atggaggcaa ttgttgaaca ggaatcatca atacaaggat atgacatgac caaagcctgt
     2761 ttcaagggag acagagtaaa tagccccaaa actttcagta ttggaactca agaaggaaaa
     2821 ctagtaaaag gatcctttgg aaaagcacta agagtaatat ttactaaatg cttgatgcac
     2881 tatgtatttg gaaatgccca attggagggg tttagtgccg agtctaggag acttctactg
     2941 ttgattcaag cattaaagga cagaaagggc ccttgggtgt tcgacttaga gggaatgtat
     3001 tctggaatag aagaatgtat tagcaacaac ccttgggtaa tacagagtgt atactggttc
     3061 aatgaatggt tgggctttga gaaggagggc aacaaggttc tcgaaagtgt agacgagatt
     3121 atggacgaat aggccgccga ctacaaagac catgacggtg attataaaga tcatgacatc
     3181 gactacaagg atgacgatga caagtaggcc gctcgagcat gcatctagag ggccctattc
     3241 tatagtgtca cctaaatgct agagctcgct gatcagcctc gactgtgcct tctagttgcc
     3301 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     3361 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     3421 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     3481 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     3541 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     3601 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     3661 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     3721 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     3781 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     3841 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     3901 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     3961 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     4021 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4081 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4141 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     4201 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     4261 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat
     4321 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     4381 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     4441 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     4501 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4561 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4621 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4681 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4741 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4801 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     4861 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4921 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4981 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     5041 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     5101 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     5161 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     5221 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     5281 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     5341 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     5401 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     5461 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     5521 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     5581 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     5641 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     5701 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5761 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5821 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5881 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5941 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6001 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6061 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6121 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6181 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6241 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     6301 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6361 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6421 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6481 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6541 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6601 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6661 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6721 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6781 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6841 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6901 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6961 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     7021 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     7081 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     7141 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     7201 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     7261 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     7321 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7381 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7441 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7501 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7561 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctg
//
