LOCUS       sequence-430090-        6858 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of FLAG/HA-p54.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6858)
  AUTHORS   Nishi K, Takahashi T, Suzawa M, Miyakawa T, Nagasawa T, Ming Y, 
            Tanokura M, Ui-Tei K
  TITLE     Control of the localization and function of a miRNA silencing 
            component TNRC6A by Argonaute protein.
  JOURNAL   Nucleic Acids Res. 2015 Nov 16;43(20):9856-73. doi: 
            10.1093/nar/gkv1026. Epub 2015 Oct 7.
  PUBMED    26446993
REFERENCE   2  (bases 1 to 6858)
  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: 
            "10.1093/nar/gkv1026"; journalName: "Nucleic Acids Res"; date: 
            "2015-11-16- 16"; volume: "43"; issue: "20"; pages: "9856-73"
COMMENT     Sequence Label: pcDNA3.1-FLAG/HA-p54
FEATURES             Location/Qualifiers
     source          1..6858
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(38..57)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        229..608
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        609..812
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     763..783
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     857..876
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        857..875
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             903..926
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             927..953
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2443..2460)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2449..2673
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2719..3147
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2806..2825)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3016..3037
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3156..3176)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3161..3490
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3341..3476
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3403..3422
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3557..4351
                     /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(3611..3630)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4221..4240
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4525..4646
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4562..4581)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4616..4635
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4695..4711)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4695..4711)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4708..4730)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4719..4735
                     /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(4743..4749,4750..4767,4768..4773))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4788..4809
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4926..4943)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5097..5685)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5177..5196)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5856..6647,6648..6716))
                     /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     6479..6498
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6717..6821)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat
       61 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
      121 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg
      181 ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat
      241 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
      301 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
      361 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      421 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      481 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      541 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      601 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      661 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      721 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      781 gtgtacggtg ggaggtctat ataagcagag ctctctggct aactagagaa cccactgctt
      841 actggcttat cgaaattaat acgactcact atagggagac ccaagctggc tagcccacca
      901 tggactacaa ggacgacgat gacaagtacc cttatgacgt gcccgattac gctaagctta
      961 tgagcacggc cagaacagag aaccctgtta taatgggtct gtccagtcaa aatggtcagc
     1021 tgagaggccc tgtgaaaccc actggtggcc ctggaggagg gggcacacag acacagcaac
     1081 agatgaacca gctgaaaaac accaacacaa tcaataatgg cactcagcag caagcacaga
     1141 gtatgaccac cactattaaa cctggtgatg actggaaaaa gactttaaaa ctccctccaa
     1201 aggatctaag aatcaaaact tcggatgtga cctccacaaa aggaaatgag tttgaagatt
     1261 actgtttgaa acgggagtta ctgatgggaa tttttgaaat gggctgggaa aagccatctc
     1321 ctattcagga ggagagcatt cccattgctt tatctggtag ggatatctta gctagagcaa
     1381 aaaatggaac aggcaagagc ggtgcctacc tcattccctt acttgaacgg ctagacctga
     1441 agaaggacaa tatacaagca atggtgattg ttcccactag agaacttgct ctacaggtca
     1501 gtcaaatttg catccaggtc agcaaacaca tgggaggggc caaagtgatg gcaaccacag
     1561 gaggaaccaa tttacgagat gacataatga ggcttgatga tacagtgcac gtggtgattg
     1621 ctacccctgg gagaatcctg gatcttatta agaaaggagt agcaaaggtt gatcatgtcc
     1681 agatgatagt attggatgag gcagataagt tgctgtcaca ggattttgtg cagataatgg
     1741 aggatattat tctcacgcta cctaaaaaca ggcagatttt actatattcc gctactttcc
     1801 ctcttagtgt acagaagttc atgaattccc atttgcagaa accctatgag attaacctga
     1861 tggaggaact aactctgaag ggagtaaccc agtactacgc atatgtaact gagcgccaaa
     1921 aagtacactg cctcaacaca cttttctcca ggcttcagat aaaccagtcg atcattttct
     1981 gtaactcctc tcagcgagtt gaattgctag ccaagaagat ttctcaactg ggttattctt
     2041 gcttctatat tcatgctaaa atgaggcagg aacatcgaaa tcgtgtattt catgatttcc
     2101 gaaatggctt atgccgcaat cttgtttgca ctgatctgtt tacccgaggt attgatatac
     2161 aagctgtgaa tgtggtaata aactttgatt tcccaaagct ggcagagacc tatctccatc
     2221 gtattggaag atcaggtcgc tttggtcatc ttggcttagc catcaacttg atcacatatg
     2281 atgatcgctt caacctgaaa agtattgagg agcagctggg aacagaaatt aaacctattc
     2341 cgagcaacat tgataagagc ctgtatgtgg cagaatacca cagcgagcct gtagaagatg
     2401 agaaacctta atctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta
     2461 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     2521 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     2581 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     2641 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     2701 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     2761 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     2821 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     2881 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     2941 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     3001 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     3061 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     3121 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     3181 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3241 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3301 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     3361 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3421 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     3481 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
     3541 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     3601 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     3661 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     3721 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     3781 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     3841 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     3901 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     3961 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     4021 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     4081 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     4141 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     4201 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     4261 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     4321 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     4381 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     4441 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     4501 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     4561 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     4621 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     4681 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     4741 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     4801 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     4861 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     4921 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     4981 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5041 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5101 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5161 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5221 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5281 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5341 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5401 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5461 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5521 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5581 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5641 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5701 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5761 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     5821 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     5881 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     5941 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6001 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6061 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6121 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6181 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6241 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6301 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6361 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6421 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6481 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6541 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6601 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6661 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     6721 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6781 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     6841 ccacctgacg tcgacgga
//
