LOCUS       sequence-244704-        8736 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8736)
  AUTHORS   Lian SL, Li S, Abadal GX, Pauley BA, Fritzler MJ, Chan EK
  TITLE     The C-terminal half of human Ago2 binds to multiple GW-rich regions 
            of GW182 and requires GW182 to mediate silencing.
  JOURNAL   RNA. 2009 May . 15(5):804-13.
  PUBMED    19324964
REFERENCE   2  (bases 1 to 8736)
  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"; journalName: "RNA. 2009 
            May . 15(5):804-13."
COMMENT     Sequence Label: FLAG-Ago2
FEATURES             Location/Qualifiers
     source          1..8736
                     /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"
     CDS             614..679
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    692..716
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      2678..2687
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3355..3379)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3386..3415
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    3422..4044
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3621..3640)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(4068..4088)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5544..5740
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5591..5726
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5778..5797)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5779..5797)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5811..5827)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5811..5827)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5824..5846)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5835..5851
                     /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(5859..5865,5866..5883,5884..5889))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5904..5925
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6042..6059)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6213..6801)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6293..6312)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6972..7763,7764..7832))
                     /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     7595..7614
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7833..7937)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7964..8419
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8051..8070)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8261..8282
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8545..8567
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8559..8576
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8560..8576
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
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 ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agaattaacc atggactaca aagaccatga cggtgattat aaagatcatg acatcgatta
      661 caaggatgac gatgacaagc ttgcggccat cacaagtttg tacaaaaaag caggctcttt
      721 aaaggaacca attcagtcga ctggatccat gcacccattc cagtggtgta acgcacttgc
      781 acctcctgcg ccgccgcccc ccatccaagg atatgccttc aagcctccac ctagacccga
      841 ctttgggacc tccgggagaa caatcaaatt acaggccaat ttcttcgaaa tggacatccc
      901 caaaattgac atctatcatt atgaattgga tatcaagcca gagaagtgcc cgaggagagt
      961 taacagggaa atcgtggaac acatggtcca gcactttaaa acacagatct ttggggatcg
     1021 gaagcccgtg tttgacggca ggaagaatct atacacagcc atgccccttc cgattgggag
     1081 ggacaaggtg gagctggagg tcacgctgcc aggagaaggc aaggatcgca tcttcaaggt
     1141 gtccatcaag tgggtgtcct gcgtgagctt gcaggcgtta cacgatgcac tttcagggcg
     1201 gctgcccagc gtcccttttg agacgatcca ggccctggac gtggtcatga ggcacttgcc
     1261 atccatgagg tacacccccg tgggccgctc cttcttcacc gcgtccgaag gctgctctaa
     1321 ccctcttggc gggggccgag aagtgtggtt tggcttccat cagtccgtcc ggccttctct
     1381 ctggaaaatg atgctgaata ttgatgtgtc agcaacagcg ttttacaagg cacagccagt
     1441 aatcgagttt gtttgtgaag ttttggattt taaaagtatt gaagaacaac aaaaacctct
     1501 gacagattcc caaagggtaa agtttaccaa agaaattaaa ggtctaaagg tggagataac
     1561 gcactgtggg cagatgaaga ggaagtaccg tgtctgcaat gtgacccggc ggcccgccag
     1621 tcaccaaaca ttcccgctgc agcaggagag cgggcagacg gtggagtgca cggtggccca
     1681 gtatttcaag gacaggcaca agttggttct gcgctacccc cacctcccat gtttacaagt
     1741 cggacaggag cagaaacaca cctaccttcc cctggaggtc tgtaacattg tggcaggaca
     1801 aagatgtatt aaaaaattaa cggacaatca gacctcaacc atgatcagag caactgctat
     1861 gtcggcgccc gatcggcaag aagagattag caaattgatg cgaagtgcaa gtttcaacac
     1921 agatccatac gtccgtgaat ttggaatcat ggtcaaagat gagatgacag acgtgactgg
     1981 gcgggtgctg cagccgccct ccatcctcta cgggggcagg aataaagcta ttgcgacccc
     2041 tgtccagggc gtctgggaca tgcggaacaa gcagttccac acgggcatcg agatcaaggt
     2101 gtgggccatt gcgtgcttcg ccccccagcg ccagtgcacg gaagtccatc tgaagtcctt
     2161 cacagagcag ctcagaaaga tctcgagaga cgctggcatg cccatccagg gccagccgtg
     2221 cttctgcaaa tacgcgcagg gggcggacag cgtggagccc atgttccggc acctgaagaa
     2281 cacgtatgcg ggcctgcagc tggtggtggt catcctgccc ggcaagacgc ccgtgtacgc
     2341 cgaggtcaag cgcgtgggag acacggtgct ggggatggcc acgcagtgcg tgcagatgaa
     2401 gaacgtgcag aggaccacgc cacagaccct gtccaacctc tgcctgaaga tcaacgtcaa
     2461 gctgggaggc gtgaacaaca tcctgctgcc ccagggcagg ccgccggtgt tccagcagcc
     2521 cgtcatcttt ctgggagcag acgtcactca cccccccgcc ggggatggga agaagccctc
     2581 cattgccgcc gtggtgggca gcatggacgc ccaccccaat cgctactgcg ccaccgtgcg
     2641 cgtgcagcag caccggcagg agatcataca agacctggcc gccatggtcc gcgagctcct
     2701 catccagttc tacaagtcca cgcgcttcaa gcccacccgc atcatcttct accgcgacgg
     2761 tgtctctgaa ggccagttcc agcaggttct ccaccacgag ttgctggcca tccgtgaggc
     2821 ctgtatcaag ctagaaaaag actaccagcc cgggatcacc ttcatcgtgg tgcagaagag
     2881 gcaccacacc cggctcttct gcactgacaa gaacgagcgg gttgggaaaa gtggaaacat
     2941 tccagcaggc acgactgtgg acacgaaaat cacccacccc accgagttcg acttctacct
     3001 gtgtagtcac gctggcatcc aggggacaag caggccttcg cactatcacg tcctctggga
     3061 cgacaatcgt ttctcctctg atgagctgca gatcctaacc taccagctgt gtcacaccta
     3121 cgtgcgctgc acacgctccg tgtccatccc agcgccagca tactacgctc acctggtggc
     3181 cttccgggcc aggtaccacc tggtggataa ggaacatgac agtgctgaag gaagccatac
     3241 ctctgggcag agtaacgggc gagaccacca agcactggcc aaggcggtcc aggttcacca
     3301 agacactctg cgcaccatgt actttgctta agcggccgca ctcgagatat ctagacccag
     3361 ctttcttgta caaagtggtg attccgaaca aaaactcatc tcagaagagg atctgtgacc
     3421 cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc
     3481 agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc
     3541 cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac
     3601 aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg
     3661 gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt
     3721 gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg
     3781 gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc
     3841 ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct
     3901 gattttaaaa taactatacc agcaggagga cgtccagaca cagcataggc tacctggcca
     3961 tgcccaaccg gtgggacatt tgagttgctt gcttggcact gtcctctcat gcgttgggtc
     4021 cactcagtag atgcctgttg aattgggtac gcggccagct tggctgtgga atgtgtgtca
     4081 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4141 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4201 aagcactgat gaatccccta atgattttgg taaaaatcat taagttaagg tggatacaca
     4261 tcttgtcata tgatcaaatg gtttcgcgaa aaatcaataa tcagacaaca agatgtgcga
     4321 actcgatatt ttacacgact ctctttacca attctgcccc gaattacact taaaacgact
     4381 caacagctta acgttggctt gccacgcctt acttgactgt aaaactctca ctcttaccga
     4441 acttggccgt aacctgccaa ccaaagcgag aacaaaacat aacatcaaac gaatcgaccg
     4501 attgttaggt aatcgtcacc tccacaaaga gcgactcgct gtataccgtt ggcatgctag
     4561 ctttatctgt tcgggcaata cgatgcccat tgtacttgtt gactggtctg atattcgtga
     4621 gcaaaaacga cttatggtat tgcgagcttc agtcgcacta cacggtcgtt ctgttactct
     4681 ttatgagaaa gcgttcccgc tttcagagca atgttcaaag aaagctcatg accaatttct
     4741 agccgacctt gcgagcattc taccgagtaa caccacaccg ctcattgtca gtgatgctgg
     4801 ctttaaagtg ccatggtata aatccgttga gaagctgggt tggtactggt taagtcgagt
     4861 aagaggaaaa gtacaatatg cagacctagg agcggaaaac tggaaaccta tcagcaactt
     4921 acatgatatg tcatctagtc actcaaagac tttaggctat aagaggctga ctaaaagcaa
     4981 tccaatctca tgccaaattc tattgtataa atctcgctct aaaggccgaa aaaatcagcg
     5041 ctcgacacgg actcattgtc accacccgtc acctaaaatc tactcagcgt cggcaaagga
     5101 gccatgggtt ctagcaacta acttacctgt tgaaattcga acacccaaac aacttgttaa
     5161 tatctattcg aagcgaatgc agattgaaga aaccttccga gacttgaaaa gtcctgccta
     5221 cggactaggc ctacgccata gccgaacgag cagctcagag cgttttgata tcatgctgct
     5281 aatcgccctg atgcttcaac taacatgttg gcttgcgggc gttcatgctc agaaacaagg
     5341 ttgggacaag cacttccagg ctaacacagt cagaaatcga aacgtactct caacagttcg
     5401 cttaggcatg gaagttttgc ggcattctgg ctacacaata acaagggaag acttactcgt
     5461 ggctgcaacc ctactagctc aaaatttatt cacacatggt tacgctttgg ggaaattatg
     5521 aggggatctc tcagtgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     5581 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     5641 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     5701 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcctcg aggaactgaa
     5761 aaaccagaaa gttaattccc tatagtgagt cgtattaaat tcgtaatcat gtcatagctg
     5821 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     5881 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     5941 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     6001 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     6061 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     6121 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     6181 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6241 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     6301 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     6361 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     6421 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6481 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6541 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6601 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     6661 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6721 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6781 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6841 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6901 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6961 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     7021 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     7081 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     7141 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7201 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     7261 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     7321 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     7381 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     7441 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     7501 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     7561 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     7621 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     7681 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     7741 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     7801 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     7861 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     7921 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc
     7981 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     8041 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     8101 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     8161 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     8221 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     8281 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     8341 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     8401 atattaacgc ttacaatttc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     8461 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     8521 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca
     8581 agctgatcta tacattgaat caatattggc aattagccat attagtcatt ggttatatag
     8641 cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca
     8701 tttatattgg ctcatgtcca atatgaccgc catgtt
//
