LOCUS       sequence-367583-        8065 bp    DNA     circular SYN 15-APR-2025
DEFINITION  argonaute RISC Catalytic Component 1 (AGO1 Human).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8065)
  AUTHORS   Briskin D, Wang PY, Bartel DP
  TITLE     The biochemical basis for the cooperative action of microRNAs.
  JOURNAL   Proc Natl Acad Sci U S A. 2020 Jul 28;117(30):17764-17774. doi: 
            10.1073/pnas.1920404117. Epub 2020 Jul 13.
  PUBMED    32661162
REFERENCE   2  (bases 1 to 8065)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1073/pnas.1920404117"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2020-07-28- 28"; volume: "117"; issue: "30"; pages: 
            "17764-17774"
COMMENT     Sequence Label: pcDNA3.3-3XFLAG-AGO1
FEATURES             Location/Qualifiers
     source          1..8065
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        50..429
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        430..633
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     584..604
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     630..654
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      742..751
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             751..816
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3451..3470)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3495..3543
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3745..4173
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3832..3851)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4042..4063
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4182..4202)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4187..4516
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4367..4502
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4429..4448
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4583..5377
                     /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(4637..4656)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5247..5266
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5553..5674
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5590..5609)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5644..5663
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5723..5739)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5723..5739)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5736..5758)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5747..5763
                     /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(5771..5777,5778..5795,5796..5801))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5816..5837
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5954..5971)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6125..6713)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6205..6224)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6884..7675,7676..7744))
                     /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     7507..7526
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7745..7849)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7924..7943)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
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 gtggttctga agcgggaccc
      841 tcgggagcag ctgcgggcgc ttacctgccc cccctgcagc aggtgttcca ggcacctcgc
      901 cggcctggca ttggcactgt ggggaaacca atcaagctcc tggccaatta ctttgaggtg
      961 gacatcccta agatcgacgt gtaccactac gaggtggaca tcaagccgga taagtgtccc
     1021 cgtagagtca accgggaagt ggtggaatac atggtccagc atttcaagcc tcagatcttt
     1081 ggtgatcgca agcctgtgta tgatggaaag aagaacattt acactgtcac agcactgccc
     1141 attggcaacg aacgggtcga ctttgaggtg acaatccctg gggaagggaa ggatcgaatc
     1201 tttaaggtct ccatcaagtg gctagccatt gtgagctggc gaatgctgca tgaggccctg
     1261 gtcagcggcc agatccctgt tcccttggag tctgtgcaag ccctggatgt ggccatgagg
     1321 cacctggcat ccatgaggta cacccctgtg ggccgctcct tcttctcacc gcctgagggc
     1381 tactaccacc cgctgggggg tgggcgcgag gtctggttcg gctttcacca gtctgtgcgc
     1441 cctgccatgt ggaagatgat gctcaacatt gatgtctcag ccactgcctt ttataaggca
     1501 cagccagtga ttgagttcat gtgtgaggtg ctggacatca ggaacataga tgagcagccc
     1561 aagcccctca cggactctca gcgcgttcgc ttcaccaagg agatcaaggg cctgaaggtg
     1621 gaagtcaccc actgtggaca gatgaagagg aagtaccgcg tgtgtaatgt tacccgtcgc
     1681 cctgctagcc atcagacatt ccccttacag ctggagagtg gacagactgt ggagtgcaca
     1741 gtggcacagt atttcaagca gaaatataac cttcagctca agtatcccca tctgccctgc
     1801 ctacaagttg gccaggaaca aaagcatacc taccttcccc tagaggtctg taacattgtg
     1861 gctgggcagc gctgtattaa aaagctgacc gacaaccaga cctcgaccat gataaaggcc
     1921 acagctagat ccgctccaga cagacaggag gagatcagtc gcctgatgaa gaatgccagc
     1981 tacaacttag atccctacat ccaggaattt gggatcaaag tgaaggatga catgacggag
     2041 gtgacagggc gagtgctgcc ggcgcccatc ttgcagtacg gcggccggaa ccgggccatt
     2101 gccacaccca atcagggtgt ctgggacatg cgggggaaac agttctacaa tgggattgag
     2161 atcaaagtct gggccatcgc ctgcttcgca ccccaaaaac agtgtcgaga agaggtgctc
     2221 aagaacttca cagaccagct gcggaagatt tccaaggatg cggggatgcc tatccagggt
     2281 caaccttgtt tctgcaaata tgcacagggg gcagacagcg tggagcctat gttccggcat
     2341 ctcaagaaca cctactcagg gctgcagctc attattgtca tcctgccagg gaagacgccg
     2401 gtgtatgctg aggtgaaacg tgtcggagat acactcttgg gaatggctac gcagtgtgtg
     2461 caggtgaaga acgtggtcaa gacctcacct cagactctgt ccaacctctg cctcaagatc
     2521 aatgtcaaac ttggtggcat taacaacatc ctagtcccac accagcgctc tgccgttttt
     2581 caacagccag tgatattcct gggagcagat gttacacacc ccccagcagg ggatgggaaa
     2641 aaaccttcta tcacagcagt ggtaggcagt atggatgccc accccagccg atactgtgct
     2701 actgtgcggg tacagcgacc acggcaagag atcattgaag acttgtccta catggtgcgt
     2761 gagctcctca tccaattcta caagtccacc cgtttcaagc ctacccgcat catcttctac
     2821 cgagatgggg tgcctgaagg ccagctaccc cagatactcc actatgagct actggccatt
     2881 cgtgatgcct gcatcaaact ggaaaaggac taccagcctg ggatcactta tattgtggtg
     2941 cagaaacgcc atcacacccg ccttttctgt gctgacaaga atgagcgaat tgggaagagt
     3001 ggtaacatcc cagctgggac cacagtggac accaacatca cccacccatt tgagtttgac
     3061 ttctatctgt gcagccacgc aggcatccag ggcaccagcc gaccatccca ttactatgtt
     3121 ctttgggatg acaaccgttt cacagcagat gagctccaga tcctgacgta ccagctgtgc
     3181 cacacttacg tacgatgcac acgctctgtc tctatcccag cacctgccta ctatgcccgc
     3241 ctggtggctt tccgggcacg ataccacctg gtggacaagg agcatgacag tggagagggg
     3301 agccacatat cggggcagag caatgggcgg gacccccagg ccctggccaa agccgtgcag
     3361 gttcaccagg atactctgcg caccatgtac ttcgcttgaa agggttcgat ccctaccggt
     3421 tagtaatgag tttaaacggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     3481 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg
     3541 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     3601 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     3661 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagca gatctgcgca
     3721 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3781 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     3841 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3901 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3961 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     4021 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     4081 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     4141 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     4201 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4261 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4321 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     4381 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     4441 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4501 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
     4561 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     4621 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4681 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4741 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     4801 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4861 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4921 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4981 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     5041 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     5101 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     5161 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     5221 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     5281 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     5341 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa
     5401 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     5461 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5521 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     5581 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5641 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     5701 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5761 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5821 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5881 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5941 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     6001 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6061 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6121 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6181 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6241 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6301 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6361 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6421 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6481 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6541 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6601 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6661 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6721 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6781 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6841 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6901 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6961 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     7021 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7081 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7141 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7201 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7261 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7321 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7381 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7441 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7501 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7561 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7621 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7681 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7741 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7801 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7861 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
     7921 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     7981 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
     8041 caattgcatg aagaatctgc ttagg
//
