LOCUS       sequence-181373-        5514 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of NS1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5514)
  AUTHORS   Huang X, Zheng M, Wang P, Mok BW, Liu S, Lau SY, Chen P, Liu YC, Liu
            H, Chen Y, Song W, Yuen KY, Chen H
  TITLE     An NS-segment exonic splicing enhancer regulates influenza A virus 
            replication in mammalian cells.
  JOURNAL   Nat Commun. 2017 Mar 21;8:14751. doi: 10.1038/ncomms14751.
  PUBMED    28323816
REFERENCE   2  (bases 1 to 5514)
  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: "Nat 
            Commun."; date: "2017-03-21"; volume: "8:14751. doi"; pages: " 
            10.1038/ncomms14751"
COMMENT     Sequence Label: pCX-V5-H7N9-NS1
FEATURES             Location/Qualifiers
     source          1..5514
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1594..1616
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             2393..2434
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2538..2557)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2603..2658
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2657..2676)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3017..3033)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3017..3033)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3030..3052)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3041..3057
                     /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(3065..3071,3072..3089,3090..3095))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3110..3131
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3190..3385
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3236..3371
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3391..3525
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3428..3447)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3482..3501
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3593..3610)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3764..4352)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3844..3863)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4523..5314,5315..5383))
                     /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     5146..5165
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5384..5488)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tgttgtgctg tctcatcatt ttggcaaaga attcgcggcc gcatggattc
     1741 caatactgtg tcaagcttcc aggtagactg ctttctttgg catgtccgca aacgatttgc
     1801 agaccaagaa atgggtgatg ccccatttct agaccggctt cgccgagatc agaagtccct
     1861 gagaggaaga agcagcactc ttggtctgga catcagaact gccacgcgtg aaggaaagca
     1921 tatagtggag cggattttag aggaagagtc agatgaagca tttaaaatga gtattgcttc
     1981 agtgccagct ccacgctatc taactgacat gactcttgaa gaaatgtcaa gagattggtt
     2041 aatgctcatt cccaaacaga aaataacagg gtccctatgc attagaatgg accaagcaat
     2101 agtggacaaa aacatcacat tgaaagcaaa tttcagtgtg attttcaatc ggcttgaagc
     2161 cctgatacta cttagagctt ttacggaaga aggagcaatt gtaggcgaaa tctcaccatt
     2221 accttctctt ccaggacata ctgacaagga tgtcaaaaat gcaattgaga tcctcatcgg
     2281 aggatttgaa tggaatgata acacagttcg agtctctgaa actctacaga gattcgcttg
     2341 gagaagcagc gatgaggatg ggagatctcc actctctaca aagctcgaga tgggtaagcc
     2401 tatccctaac cctctcctcg gtctcgattc tacgtaggta ccatcgatac cggtgaattc
     2461 actcctcagg tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa tgccctggct
     2521 cacaaatacc actgagatct ttttccctct gccaaaaatt atggggacat catgaagccc
     2581 cttgagcatc tgacttctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg
     2641 aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt aaaacatcag
     2701 aatgagtatt tggtttagag tttggcaaca tatgccatat gctggctgcc atgaacaaag
     2761 gtggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca
     2821 tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc
     2881 tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg
     2941 actactccca gtcatagctg tccctcttct cttatgaaga tccctcgacc tgcagcccaa
     3001 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     3061 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     3121 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     3181 agcggatccg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3241 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3301 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     3361 ttttggaggc ctaggctttt gcaaaaagct aacttgttta ttgcagctta taatggttac
     3421 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     3481 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatccgctg cattaatgaa
     3541 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     3601 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3661 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3721 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3781 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3841 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3901 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat
     3961 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4021 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4081 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4141 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4201 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4261 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4321 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4381 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4441 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4501 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4561 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4621 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4681 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     4741 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     4801 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     4861 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     4921 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     4981 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5041 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5101 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5161 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5221 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5281 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5341 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5401 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5461 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctg
//
