LOCUS       sequence-336041-        6944 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of the Nucleoprotein of the A/New 
            Caledonia/20/99 (H1N1) Influenza virus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6944)
  AUTHORS   Di Pietro A, Kajaste-Rudnitski A, Oteiza A, Nicora L, Towers GJ, 
            Mechti N, Vicenzi E
  TITLE     TRIM22 inhibits influenza A virus infection by targeting the viral 
            nucleoprotein for degradation.
  JOURNAL   J Virol. 2013 Apr;87(8):4523-33. doi: 10.1128/JVI.02548-12. Epub 
            2013 Feb 13.
  PUBMED    23408607
REFERENCE   2  (bases 1 to 6944)
  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"; doi: 
            "10.1128/JVI.02548-12"; journalName: "J Virol"; date: "2013-04"; 
            volume: "87"; issue: "8"; pages: "4523-33"
COMMENT     Sequence Label: pcDNA3-NP
FEATURES             Location/Qualifiers
     source          1..6944
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      733..742
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2263..2281)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2264..2281)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2301..2318)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2307..2531
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2577..3005
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2664..2683)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2874..2895
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3014..3034)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3019..3348
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3199..3334
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3261..3280
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3415..4209
                     /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(3469..3488)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4079..4098
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4383..4504
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4420..4439)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4474..4493
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4553..4569)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4553..4569)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4566..4588)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4577..4593
                     /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(4601..4607,4608..4625,4626..4631))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4646..4667
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4784..4801)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4955..5543)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5035..5054)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5714..6505,6506..6574))
                     /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     6337..6356
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6575..6679)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6754..6773)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatccac tagtaacggc cgccagtgtg ctggaattct gcagatatcc
      721 atcacactgg cggccgccat ggcgtcccaa ggcaccaaac ggtcttacga acagatggag
      781 actgatgggg aacgccagaa tgcaactgaa atcagagcat ccgtcggaag aatgattggt
      841 ggaattgggc gattctacat ccaaatgtgc accgagctta aactcaatga ttatgaggga
      901 cgactgatcc agaacagctt gacaatagag agaatggtgc tctctgcttt tgatgagagg
      961 aggaataaat atctggaaga acatcccagc gcggggaaag atcctaagaa aactggagga
     1021 cccatataca agagagtaga tggaaagtgg gtgagggaac tcgtccttta tgacaaagaa
     1081 gaaataaggc ggatttggcg ccaagccaac aatggtgatg atgcaacggc tggtttgact
     1141 cacattatga tctggcattc taatttgaat gatacaactt accagaggac aagagctctt
     1201 gtccgcaccg gaatggatcc caggatgtgc tctttgatgc aaggttcaac tctccctaga
     1261 agatctggag cagcaggcgc tgcagtcaaa ggagttggga caatggtgtt ggagttaatc
     1321 aggatgatca aacgtgggat caatgaccga aacttctgga ggggtgagaa tggaagaaaa
     1381 acaaggattg cttatgagag aatgtgcaac attctcaaag gaaaatttca aacagctgca
     1441 caaaaagcaa tgatggatca agtgagagaa agccggaacc caggaaatgc tgagatcgaa
     1501 gatctcactt ttctggcacg gtctgcactc atattaagag ggtcagttgc tcacaagtct
     1561 tgcctgcctg cctgtgtgta tggaccagcc gtagccagtg ggtacgactt cgaaaaagag
     1621 ggatactctt tggtaggggt agaccctttt aaactgcttc aaaccagtca ggtatacagc
     1681 ctaatcagac caaacgagaa tcccgcacac aagagtcagt tggtgtggat ggcatgcaat
     1741 tctgctgcat ttgaagatct aagagtgtca agcttcatca gagggacaag agtacttcca
     1801 agggggaagc tctccactag aggagtacaa attgcttcaa atgaaaacat ggatgctatt
     1861 gtatcaagta ctcttgaact gagaagcaga tactgggcca taagaaccag aagtggaggg
     1921 aacactaatc aacaaagggc ctctgcgggc caaatcagca cacaacctac gttttctgtg
     1981 cagagaaacc tcccatttga caaaacaacc atcatggcag cattcactgg gaatacggag
     2041 ggaagaacat cagacatgag ggcagaaatc ataaagatga tggaaagtgc aagaccagaa
     2101 gaagtgtcct tccaggggcg gggagtcttt gagctctcgg acgaaagggc aacgaacccg
     2161 atcgtgccct cctttgacat gagtaatgaa ggatcttatt tcttcggaga caatgcagag
     2221 gagtacgaca attaactcga gcatgcatct agagggccct attctatagt gtcacctaaa
     2281 tgctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     2341 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     2401 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     2461 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     2521 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     2581 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2641 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2701 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     2761 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     2821 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     2881 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     2941 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3001 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3061 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3121 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3181 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3241 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     3301 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     3361 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3421 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3481 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3541 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     3601 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     3661 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     3721 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     3781 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     3841 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     3901 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     3961 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4021 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4081 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4141 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4201 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     4261 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4321 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     4381 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     4441 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     4501 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     4561 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     4621 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     4681 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     4741 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     4801 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     4861 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4921 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4981 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5041 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5101 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5161 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5221 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5281 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5341 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     5401 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5461 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5521 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5581 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5641 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     5701 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5761 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     5821 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     5881 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     5941 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6001 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6061 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6121 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6181 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6241 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6301 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6361 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6421 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6481 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6541 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     6601 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     6661 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     6721 gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     6781 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
     6841 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
     6901 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgtt
//
