LOCUS       sequence-78267-e        8896 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 8896)
  AUTHORS   Lee Y, Hur I, Park SY, Kim YK, Suh MR, Kim VN
  TITLE     The role of PACT in the RNA silencing pathway.
  JOURNAL   EMBO J. 2006 Feb 8;25(3):522-32. Epub 2006 Jan 19.
  PUBMED    16424907
REFERENCE   2  (bases 1 to 8896)
  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: "EMBO J."; 
            date: "2006-02-8"; volume: "25(3)"; pages: "522-32. Epub 2006 Jan 
            19"
COMMENT     Sequence Label: pcDNA3.1-FLAG-Dicer-delDUF
FEATURES             Location/Qualifiers
     source          1..8896
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             909..932
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(4490..4507)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4496..4720
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4766..5194
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4853..4872)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5063..5084
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5203..5223)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5208..5537
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5388..5523
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5450..5469
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5604..6398
                     /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(5658..5677)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6268..6287
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6572..6693
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6609..6628)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6663..6682
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6742..6758)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6742..6758)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6755..6777)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6766..6782
                     /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(6790..6796,6797..6814,6815..6820))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6835..6856
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6973..6990)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7144..7729)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7224..7243)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7900..8691,8692..8760))
                     /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     8523..8542
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8761..8865)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 ccaccatgga ctacaaggac gacgatgaca agtaagctta ttccagagtg tttgagggat
      961 agttatccca gacctgatca gccctgttac ctgtatgtga taggaatggt tttaactaca
     1021 cctttacctg atgaactcaa ctttagaagg cggaagctct atcctcctga agataccaca
     1081 agatgctttg gaatactgac ggccaaaccc atacctcaga ttccacactt tcctgtgtac
     1141 acacgctctg gagaggttac catatccatt gagttgaaga agtctggttt catgttgtct
     1201 ctacaaatgc ttgagttgat tacaagactt caccagtata tattctcaca tattcttcgg
     1261 cttgaaaaac ctgcactaga atttaaacct acagacgctg attcagcata ctgtgttcta
     1321 cctcttaatg ttgttaatga ctccagcact ttggatattg actttaaatt catggaagat
     1381 attgagaagt ctgaagctcg cataggcatt cccagtacaa agtatacaaa agaaacaccc
     1441 tttgttttta aattagaaga ttaccaagat gccgttatca ttccaagata tcgcaatttt
     1501 gatcagcctc atcgatttta tgtagctgat gtgtacactg atcttacccc actcagtaaa
     1561 tttccttccc ctgagtatga aacttttgca gaatattata aaacaaagta caaccttgac
     1621 ctaaccaatc tcaaccagcc actgctggat gtggaccaca catcttcaag acttaatctt
     1681 ttgacacctc gacatttgaa tcagaagggg aaagcgcttc ctttaagcag tgctgagaag
     1741 aggaaagcca aatgggaaag tctgcagaat aaacagatac tggttccaga actctgtgct
     1801 atacatccaa ttccagcatc actgtggaga aaagctgttt gtctccccag catactttat
     1861 cgccttcact gccttttgac tgcagaggag ctaagagccc agactgccag cgatgctggc
     1921 gtgggagtca gatcacttcc tgcggatttt agatacccta acttagactt cgggtggaaa
     1981 aaatctattg acagcaaatc tttcatctca atttctaact cctcttcagc tgaaaatgat
     2041 aattactgta agcacagcac aattgtccct gaaaatgctg cacatcaagg tgctaataga
     2101 acctcctctc tagaaaatca tgaccaaatg tctgtgaact gcagaacgtt gctcagcgag
     2161 tcccctggta agctccacgt tgaagtttca gcagatctta cagcaattaa tggtctttct
     2221 tacaatcaaa atctcgccaa tggcagttat gatttagcta acagagactt ttgccaagga
     2281 aatcagctaa attactacaa gcaggaaata cccgtgcaac caactacctc atattccatt
     2341 cagaatttat acagttacga gaaccagccc cagcccagcg atgaatgtac tctcctgagt
     2401 aataaatacc ttgatggaaa tgctaacaaa tctacctcag atggaagtcc tgtgatggcc
     2461 gtaatgcctg gtacgacaga cactattcaa gtgctcaagg gcaggatgga ttctgagcag
     2521 agcccttcta ttgggtactc ctcaaggact cttggcccca atcctggact tattcttcag
     2581 gctttgactc tgtcaaacgc tagtgatgga tttaacctgg agcggcttga aatgcttggc
     2641 gactcctttt taaagcatgc catcaccaca tatctatttt gcacttaccc tgatgcgcat
     2701 gagggccgcc tttcatatat gagaagcaaa aaggtcagca actgtaatct gtatcgcctt
     2761 ggaaaaaaga agggactacc cagccgcatg gtggtgtcaa tatttgatcc ccctgtgaat
     2821 tggcttcctc ctggttatgt agtaaatcaa gacaaaagca acacagataa atgggaaaaa
     2881 gatgaaatga caaaagactg catgctggcg aatggcaaac tggatgagga ttacgaggag
     2941 gaggatgagg aggaggagag cctgatgtgg agggctccga aggaagaggc tgactatgaa
     3001 gatgatttcc tggagtatga tcaggaacat atcagattta tagataatat gttaatgggg
     3061 tcaggagctt ttgtaaagaa aatctctctt tctccttttt caaccactga ttctgcatat
     3121 gaatggaaaa tgcccaaaaa atcctcctta ggtagtatgc cattttcatc agattttgag
     3181 gattttgact acagctcttg ggatgcaatg tgctatctgg atcctagcaa agctgttgaa
     3241 gaagatgact ttgtggtggg gttctggaat ccatcagaag aaaactgtgg tgttgacacg
     3301 ggaaagcagt ccatttctta cgacttgcac actgagcagt gtattgctga caaaagcata
     3361 gcggactgtg tggaagccct gctgggctgc tatttaacca gctgtgggga gagggctgct
     3421 cagcttttcc tctgttcact ggggctgaag gtgctcccgg taattaaaag gactgatcgg
     3481 gaaaaggccc tgtgccctac tcgggagaat ttcaacagcc aacaaaagaa cctttcagtg
     3541 agctgtgctg ctgcttctgt ggccagttca cgctcttctg tattgaaaga ctcggaatat
     3601 ggttgtttga agattccacc aagatgtatg tttgatcatc cagatgcaga taaaacactg
     3661 aatcacctta tatcggggtt tgaaaatttt gaaaagaaaa tcaactacag attcaagaat
     3721 aaggcttacc ttctccaggc ttttacacat gcctcctacc actacaatac tatcactgat
     3781 tgttaccagc gcttagaatt cctgggagat gcgattttgg actacctcat aaccaagcac
     3841 ctttatgaag acccgcggca gcactccccg ggggtcctga cagacctgcg gtctgccctg
     3901 gtcaacaaca ccatctttgc atcgctggct gtaaagtacg actaccacaa gtacttcaaa
     3961 gctgtctctc ctgagctctt ccatgtcatt gatgactttg tgcagtttca gcttgagaag
     4021 aatgaaatgc aaggaatgga ttctgagctt aggagatctg aggaggatga agagaaagaa
     4081 gaggatattg aagttccaaa ggccatgggg gatatttttg agtcgcttgc tggtgccatt
     4141 tacatggata gtgggatgtc actggagaca gtctggcagg tgtactatcc catgatgcgg
     4201 ccactaatag aaaagttttc tgcaaatgta ccccgttccc ctgtgcgaga attgcttgaa
     4261 atggaaccag aaactgccaa atttagcccg gctgagagaa cttacgacgg gaaggtcaga
     4321 gtcactgtgg aagtagtagg aaaggggaaa tttaaaggtg ttggtcgaag ttacaggatt
     4381 gccaaatctg cagcagcaag aagagccctc cgaagcctca aagctaatca acctcaggtt
     4441 cccaatagct gagtcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg
     4501 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     4561 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     4621 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     4681 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     4741 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     4801 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     4861 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4921 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4981 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     5041 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     5101 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     5161 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     5221 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     5281 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     5341 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     5401 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     5461 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     5521 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     5581 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     5641 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     5701 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     5761 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     5821 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     5881 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     5941 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     6001 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     6061 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     6121 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     6181 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     6241 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     6301 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     6361 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     6421 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     6481 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     6541 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     6601 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     6661 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     6721 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     6781 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     6841 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     6901 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6961 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     7021 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     7081 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     7141 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     7201 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     7261 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     7321 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     7381 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     7441 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     7501 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     7561 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     7621 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     7681 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     7741 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     7801 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     7861 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     7921 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     7981 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     8041 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     8101 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     8161 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     8221 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     8281 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     8341 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     8401 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     8461 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     8521 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     8581 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     8641 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     8701 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     8761 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     8821 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     8881 agtgccacct gacgtc
//
