LOCUS       pcDNA3.3-3xFLAG-        8071 bp DNA     circular SYN 18-JUN-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8071)
  AUTHORS   PYW
  TITLE     Direct Submission
  JOURNAL   Exported Apr 27, 2024 from SnapGene 7.2.0
            https://www.snapgene.com
COMMENT     Sequence Label: pcDNA3.3-3xFLAG-hAGO2
FEATURES             Location/Qualifiers
     source          1..8071
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        47..725
                     /label=CMV promoter
                     /note="CMV promoter"
     RBS             742..751
                     /label=Kozak sequence
                     /note="Kozak"
     CDS             748..816
                     /codon_start=1
                     /label=3XFLAG peptide
                     /note="3XFLAG peptide"
                     /translation="MDYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    802..819
                     /label=Enterokinase cleavage recognition
                     /note="Enterokinase cleavage recognition sequence"
     CDS             820..828
                     /codon_start=1
                     /label=GGS linker
                     /note="Linker region"
                     /translation="GGS"
     CDS             829..3405
                     /codon_start=1
                     /label=hAGO2_S387D
                     /note="Ago2"
                     /translation="YSGAGPALAPPAPPPPIQGYAFKPPPRPDFGTSGRTIKLQANFFE
                     MDIPKIDIYHYELDIKPEKCPRRVNREIVEHMVQHFKTQIFGDRKPVFDGRKNLYTAMP
                     LPIGRDKVELEVTLPGEGKDRIFKVSIKWVSCVSLQALHDALSGRLPSVPFETIQALDV
                     VMRHLPSMRYTPVGRSFFTASEGCSNPLGGGREVWFGFHQSVRPSLWKMMLNIDVSATA
                     FYKAQPVIEFVCEVLDFKSIEEQQKPLTDSQRVKFTKEIKGLKVEITHCGQMKRKYRVC
                     NVTRRPASHQTFPLQQESGQTVECTVAQYFKDRHKLVLRYPHLPCLQVGQEQKHTYLPL
                     EVCNIVAGQRCIKKLTDNQTSTMIRATARSAPDRQEEISKLMRSADFNTDPYVREFGIM
                     VKDEMTDVTGRVLQPPSILYGGRNKAIATPVQGVWDMRNKQFHTGIEIKVWAIACFAPQ
                     RQCTEVHLKSFTEQLRKISRDAGMPIQGQPCFCKYAQGADSVEPMFRHLKNTYAGLQLV
                     VVILPGKTPVYAEVKRVGDTVLGMATQCVQMKNVQRTTPQTLSNLCLKINVKLGGVNNI
                     LLPQGRPPVFQQPVIFLGADVTHPPAGDGKKPSIAAVVGSMDAHPNRYCATVRVQQHRQ
                     EIIQDLAAMVRELLIQFYKSTRFKPTRIIFYRDGVSEGQFQQVLHHELLAIREACIKLE
                     KDYQPGITFIVVQKRHHTRLFCTDKNERVGKSGNIPAGTTVDTKITHPTEFDFYLCSHA
                     GIQGTSRPSHYHVLWDDNRFSSDELQILTYQLCHTYVRCTRSVSIPAPAYYAHLVAFRA
                     RYHLVDKEHDSAEGSHTSGQSNGRDHQALAKAVQVHQDTLRTMYFA"
     polyA_signal    3444..3715
                     /label=TK polyadenylation signal
                     /note="TK polyadenylation signal"
     rep_origin      3751..4179
                     /direction=RIGHT
                     /label=f1 replication origin
                     /note="f1 replication origin"
     promoter        4184..4553
                     /label=SV40 early promoter
                     /note="SV40 early promoter"
     CDS             4589..5383
                     /codon_start=1
                     /label=Neomycin Resistance Gene
                     /note="Neomycin Resistance Gene"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5559..5689
                     /label=SV40 polyadenylation signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(6072..6745)
                     /direction=LEFT
                     /label=pUC origin
                     /note="pUC origin"
     CDS             complement(6930..7750)
                     /codon_start=1
                     /label=Ampicillin (bla) resistance gene
                     /note="Ampicillin (bla) resistance gene"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNR"
     promoter        complement(7751..7849)
                     /label=bla promoter
                     /note="bla promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg
      661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg
      721 actctagagg atcgaaccct tgccaccatg gactacaaag accatgacgg tgattataaa
      781 gatcatgata tcgattacaa ggatgacgat gacaagcttg gtggttctta ctcgggagcc
      841 ggccccgcac ttgcacctcc tgcgccgccg ccccccatcc aaggatatgc cttcaagcct
      901 ccacctagac ccgactttgg gacctccggg agaacaatca aattacaggc caatttcttc
      961 gaaatggaca tccccaaaat tgacatctat cattatgaat tggatatcaa gccagagaag
     1021 tgcccgagga gagttaacag ggaaatcgtg gaacacatgg tccagcactt taaaacacag
     1081 atctttgggg atcggaagcc cgtgtttgac ggcaggaaga atctatacac agccatgccc
     1141 cttccgattg ggagggacaa ggtggagctg gaggtcacgc tgccaggaga aggcaaggat
     1201 cgcatcttca aggtgtccat caagtgggtg tcctgcgtga gcttgcaggc gttacacgat
     1261 gcactttcag ggcggctgcc tagcgtccct tttgagacga tccaggccct ggacgtggtc
     1321 atgaggcact tgccatccat gaggtacacc cccgtgggcc gctccttctt caccgcgtcc
     1381 gaaggctgct ctaaccctct tggcgggggc cgagaagtgt ggtttggctt ccatcagtcc
     1441 gtccggcctt ctctctggaa aatgatgctg aatattgatg tgtcagcaac agcgttttac
     1501 aaggcacagc cagtaatcga gtttgtttgt gaagttttgg attttaaaag tattgaagaa
     1561 caacaaaaac ctctgacaga ttcccaaagg gtaaagttta ccaaagaaat taaaggtcta
     1621 aaggtggaga taacgcactg tgggcagatg aagaggaagt accgcgtctg caatgtgacc
     1681 cggcggcccg ccagtcacca aacattcccg ctgcagcagg agagcgggca gacggtggag
     1741 tgcacggtgg cccagtattt caaggacagg cacaagcttg ttctgcgcta cccccacctc
     1801 ccatgtttac aagtcggaca ggagcagaaa cacacctacc ttcccctgga ggtctgtaac
     1861 attgtggcag gacaaagatg tattaaaaaa ttaacggaca atcagacctc aaccatgatc
     1921 agagcaactg ctaggtcggc gcccgatcgg caagaagaga ttagcaaatt gatgcgaagt
     1981 gcagatttca acacagatcc atacgtccgt gaatttggaa tcatggtcaa agatgagatg
     2041 acagacgtga ctgggcgggt gctgcagccg ccctccatcc tctacggggg caggaataaa
     2101 gctattgcga cccctgtcca gggcgtctgg gacatgcgga acaagcagtt ccacacgggc
     2161 atcgagatca aggtgtgggc cattgcgtgc ttcgcccccc agcgccagtg cacggaagtc
     2221 catctgaagt ccttcacaga gcagctcaga aagatctcga gagacgctgg catgcccatc
     2281 cagggccagc cgtgcttctg caaatacgcg cagggggcgg acagcgtgga gcccatgttc
     2341 cggcacctga agaacacgta tgcgggcctg cagctggtgg tggtcatcct gcccggcaag
     2401 acgcccgtgt acgccgaggt caagcgcgtg ggagacacgg tgctggggat ggccacgcag
     2461 tgcgtgcaga tgaagaacgt gcagaggacc acgccacaga ccctgtccaa cctctgcctg
     2521 aagatcaacg tcaagctggg aggcgtgaac aacatcctgc tgccccaggg caggccgccg
     2581 gtgttccagc agcccgtcat ctttctggga gcagacgtca ctcacccccc cgccggggat
     2641 gggaagaagc cctccattgc cgccgtggtg ggcagcatgg acgcccaccc caatcgctac
     2701 tgcgccaccg tgcgcgtgca gcagcaccgg caggagatca tacaagacct ggccgccatg
     2761 gtccgcgagc tcctcatcca gttctacaag tccacgcgct tcaagcccac ccgcatcatc
     2821 ttctaccgcg acggtgtctc tgaaggccag ttccagcagg ttctccacca cgagttgctg
     2881 gccatccgtg aggcctgtat caagctagaa aaagactacc agcccgggat caccttcatc
     2941 gtggtgcaga agaggcacca cacccggctc ttctgcactg acaagaacga gcgggttggg
     3001 aaaagtggaa acattccagc aggcacgact gtggacacga aaatcaccca ccccaccgag
     3061 ttcgacttct acctgtgtag tcacgctggc atccagggga caagcaggcc ttcgcactat
     3121 cacgtcctct gggacgacaa tcgtttctcc tctgatgagc tgcagatcct aacctaccag
     3181 ctgtgtcaca cctacgtgcg ctgcacacgc tccgtgtcca tcccagcgcc agcatactac
     3241 gctcacctgg tggccttccg ggccaggtac cacctggtgg ataaggaaca tgacagtgct
     3301 gaaggaagcc atacctctgg gcagagtaac gggcgagacc accaagcact ggccaaggcg
     3361 gtccaggttc accaagacac tctgcgcacc atgtactttg cttgaaaggg ttcgatccct
     3421 accggttagt aatgagttta aacgggggag gctaactgaa acacggaagg agacaatacc
     3481 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg ggtgttgggt
     3541 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     3601 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     3661 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcagatc
     3721 tgcgcagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     3781 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3841 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3901 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3961 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4021 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     4081 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     4141 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     4201 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4261 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4321 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     4381 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4441 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     4501 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     4561 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     4621 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     4681 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     4741 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     4801 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4861 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4921 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4981 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     5041 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     5101 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     5161 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     5221 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     5281 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     5341 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     5401 gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     5461 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     5521 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     5581 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5641 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     5701 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     5761 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     5821 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5881 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5941 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     6001 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     6061 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6121 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6181 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6241 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6301 cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg
     6361 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6421 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6481 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6541 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     6601 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6661 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6721 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6781 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     6841 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     6901 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     6961 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     7021 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7081 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7141 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7201 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7261 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7321 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7381 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     7441 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7501 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7561 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7621 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7681 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7741 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7801 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     7861 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtc
     7921 gactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg
     7981 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
     8041 gaccgacaat tgcatgaaga atctgcttag g
//
