LOCUS       sequence-156379-        7284 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human Ago2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7284)
  AUTHORS   Shen J, Xia W, Khotskaya YB, Huo L, Nakanishi K, Lim SO, Du Y, Wang 
            Y, Chang WC, Chen CH, Hsu JL, Wu Y, Lam YC, James BP, Liu X, Liu CG,
            Patel DJ, Hung MC
  TITLE     EGFR modulates microRNA maturation in response to hypoxia through 
            phosphorylation of AGO2.
  JOURNAL   Nature. 2013 May 16;497(7449):383-7. doi: 10.1038/nature12080. Epub 
            2013 May 1.
  PUBMED    23636329
REFERENCE   2  (bases 1 to 7284)
  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: 
            "10.1038/nature12080"; journalName: "Nature"; date: "2013-05-16- 
            16"; volume: "497"; issue: "7449"; pages: "383-7"
COMMENT     Sequence Label: pCMV5-Flag-Ago2
FEATURES             Location/Qualifiers
     source          1..7284
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     127..149
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     141..158
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     142..158
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        319..698
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        699..902
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     853..873
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             966..989
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      2922..2931
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3813..3832)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(4258..4278)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4263..4593
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4444..4579
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4631..4650)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4632..4650)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4665..4681)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4665..4681)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4678..4700)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4689..4705
                     /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(4713..4719,4720..4737,4738..4743))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4758..4779
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4896..4913)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5066..5654)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5146..5165)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5825..6616,6617..6685))
                     /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     6448..6467
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6686..6790)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6817..7245
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6904..6923)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7114..7135
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
       61 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
      121 ggttttccca gtcacgacgt tgtaaaacga cggccagtgc caagctgatc tatacattga
      181 atcaatattg gcaattagcc atattagtca ttggttatat agcataaatc aatattggct
      241 attggccatt gcatacgttg tatctatatc ataatatgta catttatatt ggctcatgtc
      301 caatatgacc gccatgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
      361 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      421 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      481 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      541 cccacttggc agtacatcaa gtgtatcata tgccaagtcc gccccctatt gacgtcaatg
      601 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttacgggac tttcctactt
      661 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
      721 ccaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      781 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaataacc
      841 ccgccccgtt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
      901 ctcgtttagt gaaccgtcag aattaattca gatctggtac cacgcgtatc gataagctta
      961 ccatggatta caaggatgac gacgataagg gatccatgta ctcgggagcc ggccccgcac
     1021 ttgcacctcc tgcgccgccg ccccccatcc aaggatatgc cttcaagcct ccacctagac
     1081 ccgactttgg gacctccggg agaacaatca aattacaggc caatttcttc gaaatggaca
     1141 tccccaaaat tgacatctat cattatgaat tggatatcaa gccagagaag tgcccgagga
     1201 gagttaacag ggaaatcgtg gaacacatgg tccagcactt taaaacacag atctttgggg
     1261 atcggaagcc cgtgtttgac ggcaggaaga atctatacac agccatgccc cttccgattg
     1321 ggagggacaa ggtggagctg gaggtcacgc tgccaggaga aggcaaggat cgcatcttca
     1381 aggtgtccat caagtgggtg tcctgcgtga gcttgcaggc gttacacgat gcactttcag
     1441 ggcggctgcc cagcgtccct tttgagacga tccaggccct ggacgtggtc atgaggcact
     1501 tgccatccat gaggtacacc cccgtgggcc gctccttctt caccgcgtcc gaaggctgct
     1561 ctaaccctct tggcgggggc cgagaagtgt ggtttggctt ccatcagtcc gtccggcctt
     1621 ctctctggaa aatgatgctg aatattgatg tgtcagcaac agcgttttac aaggcacagc
     1681 cagtaatcga gtttgtttgt gaagttttgg attttaaaag tattgaagaa caacaaaaac
     1741 ctctgacaga ttcccaaagg gtaaagttta ccaaagaaat taaaggtcta aaggtggaga
     1801 taacgcactg tgggcagatg aagaggaagt accgcgtctg caatgtgacc cggcggcccg
     1861 ccagtcacca aacattcccg ctgcagcagg agagcgggca gacggtggag tgcacggtgg
     1921 cccagtattt caaggacagg cacaagttgg ttctgcgcta cccccacctc ccatgtttac
     1981 aagtcggaca ggagcagaaa cacacctacc ttcccctgga ggtctgtaac attgtggcag
     2041 gacaaagatg tattaaaaaa ttaacggaca atcagacctc aaccatgatc agagcgactg
     2101 ctaggtcggc gcccgatcgg caagaagaga ttagcaaatt gatgcgaagt gcaagtttca
     2161 acacagatcc atacgtccgt gaatttggaa tcatggtcaa agatgagatg acagacgtga
     2221 ctgggcgggt gctgcagccg ccctccatcc tctacggggg caggaataaa gctattgcga
     2281 cccctgtcca gggcgtctgg gacatgcgga acaagcagtt ccacacgggc atcgagatca
     2341 aggtgtgggc cattgcgtgc ttcgcccccc agcgccagtg cacggaagtc catctgaagt
     2401 ccttcacaga gcagctcaga aagatctcga gagacgccgg catgcccatc cagggccagc
     2461 cgtgcttctg caaatacgcg cagggggcgg acagcgtgga gcccatgttc cggcacctga
     2521 agaacacgta tgcgggcctg cagctggtgg tggtcatcct gcccggcaag acgcccgtgt
     2581 acgccgaggt caagcgcgtg ggagacacgg tgctggggat ggccacgcag tgcgtgcaga
     2641 tgaagaacgt gcagaggacc acgccacaga ccctgtccaa cctctgcctg aagatcaacg
     2701 tcaagctggg aggcgtgaac aacatcctgc tgccccaggg caggccgccg gtgttccagc
     2761 agcccgtcat ctttctggga gcagacgtca ctcacccccc cgccggggat gggaagaagc
     2821 cctccattgc cgccgtggtg ggcagcatgg acgcccaccc caatcgctac tgcgccaccg
     2881 tgcgcgtgca gcagcaccgg caggagatca tacaagacct ggccgccatg gtccgcgagc
     2941 tcctcatcca gttctacaag tccacgcgct tcaagcccac ccgcatcatc ttctaccgcg
     3001 acggtgtctc tgaaggccag ttccagcagg ttctccacca cgagttgctg gccatccgtg
     3061 aggcctgtat caagctagaa aaagactacc agcccgggat caccttcatc gtggtgcaga
     3121 agaggcacca cacccggctc ttctgcactg acaagaacga gcgggttggg aaaagtggaa
     3181 acattccagc aggcacgact gtggacacga aaatcaccca ccccaccgag ttcgacttct
     3241 acctgtgtag tcacgctggc atccagggga caagcaggcc ttcgcactat cacgtcctct
     3301 gggacgacaa tcgtttctcc tctgatgagc tgcagatcct aacctaccag ctgtgtcaca
     3361 cctacgtgcg ctgcacacgc tccgtgtcca tcccagcgcc agcatactac gctcacctgg
     3421 tggccttccg ggccaggtac cacctggtgg ataaggaaca tgacagtgct gaaggaagcc
     3481 atacctctgg gcagagtaac gggcgagacc accaagcact ggccaaggcg gtccaggttc
     3541 accaagacac tctgcgcacc atgtactttg cttgagaatt cgatatcatc gattctagaa
     3601 ccctcgagca attccgtggc atccctgtga cccctcccca gtgcctctcc tggccctgga
     3661 agttgccact ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc
     3721 tgactaggtg tccttctata atattatggg gtggaggggg gtggtatgga gcaaggggca
     3781 agttgggaag acaacctgta gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt
     3841 ggcacaatct tggctcactg caatctccgc ctcctgggtt caagcgattc tcctgcctca
     3901 gcctcccgag ttgttgggat tccaggcatg catgaccagg ctcagctaat ttttgttttt
     3961 ttggtagaga cggggtttca ccatattggc caggctggtc tccaactcct aatctcaggt
     4021 gatctaccca ccttggcctc ccaaattgct gggattacag gcgtgaacca ctgctccctt
     4081 ccctgtcctt ctgattttaa aataactata ccagcaggag gacgtccaga cacagcatag
     4141 gctacctgcc atggcccaac cggtgggaca tttgagttgc ttgcttggca ctgtcctctc
     4201 atgcgttggg tccactcagt agatgcctgt tgaattgggt acgcggccag cttctgtgga
     4261 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4321 gcatgcatct caattagtca gcaaccaggt gtggaaaagt ccccaggctc cccagcaggc
     4381 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4441 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4501 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     4561 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc tcgaggaact gaaaaaccag
     4621 aaagttaatt ccctatagtg agtcgtatta aattcgtaat catggtcata gctgtttcct
     4681 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     4741 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     4801 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     4861 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     4921 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4981 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5041 cgtaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5101 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5161 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5221 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc
     5281 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5341 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5401 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5461 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     5521 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5581 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5641 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5701 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5761 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     5821 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5881 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5941 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6001 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6061 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6121 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6181 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6241 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6301 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6361 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6421 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6481 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     6541 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6601 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6661 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     6721 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     6781 gggttccgcg cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa
     6841 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     6901 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     6961 ctctaaatcg gggcatccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     7021 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     7081 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     7141 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     7201 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     7261 caaaatatta acgtttacaa tttc
//
