LOCUS       sequence-78269-e        7882 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 7882)
  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 7882)
  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-delPAZ
FEATURES             Location/Qualifiers
     source          1..7882
                     /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(3476..3493)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3482..3706
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3752..4180
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3839..3858)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4049..4070
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4189..4209)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4194..4523
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4374..4509
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4436..4455
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4590..5384
                     /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(4644..4663)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5254..5273
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5558..5679
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5595..5614)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5649..5668
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5728..5744)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5728..5744)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5741..5763)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5752..5768
                     /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(5776..5782,5783..5800,5801..5806))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5821..5842
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5959..5976)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6130..6715)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6210..6229)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6886..7677,7678..7746))
                     /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     7509..7528
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7747..7851)
                     /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 agtaagcttt accctaactt agacttcggg
      961 tggaaaaaat ctattgacag caaatctttc atctcaattt ctaactcctc ttcagctgaa
     1021 aatgataatt actgtaagca cagcacaatt gtccctgaaa atgctgcaca tcaaggtgct
     1081 aatagaacct cctctctaga aaatcatgac caaatgtctg tgaactgcag aacgttgctc
     1141 agcgagtccc ctggtaagct ccacgttgaa gtttcagcag atcttacagc aattaatggt
     1201 ctttcttaca atcaaaatct cgccaatggc agttatgatt tagctaacag agacttttgc
     1261 caaggaaatc agctaaatta ctacaagcag gaaatacccg tgcaaccaac tacctcatat
     1321 tccattcaga atttatacag ttacgagaac cagccccagc ccagcgatga atgtactctc
     1381 ctgagtaata aataccttga tggaaatgct aacaaatcta cctcagatgg aagtcctgtg
     1441 atggccgtaa tgcctggtac gacagacact attcaagtgc tcaagggcag gatggattct
     1501 gagcagagcc cttctattgg gtactcctca aggactcttg gccccaatcc tggacttatt
     1561 cttcaggctt tgactctgtc aaacgctagt gatggattta acctggagcg gcttgaaatg
     1621 cttggcgact cctttttaaa gcatgccatc accacatatc tattttgcac ttaccctgat
     1681 gcgcatgagg gccgcctttc atatatgaga agcaaaaagg tcagcaactg taatctgtat
     1741 cgccttggaa aaaagaaggg actacccagc cgcatggtgg tgtcaatatt tgatccccct
     1801 gtgaattggc ttcctcctgg ttatgtagta aatcaagaca aaagcaacac agataaatgg
     1861 gaaaaagatg aaatgacaaa agactgcatg ctggcgaatg gcaaactgga tgaggattac
     1921 gaggaggagg atgaggagga ggagagcctg atgtggaggg ctccgaagga agaggctgac
     1981 tatgaagatg atttcctgga gtatgatcag gaacatatca gatttataga taatatgtta
     2041 atggggtcag gagcttttgt aaagaaaatc tctctttctc ctttttcaac cactgattct
     2101 gcatatgaat ggaaaatgcc caaaaaatcc tccttaggta gtatgccatt ttcatcagat
     2161 tttgaggatt ttgactacag ctcttgggat gcaatgtgct atctggatcc tagcaaagct
     2221 gttgaagaag atgactttgt ggtggggttc tggaatccat cagaagaaaa ctgtggtgtt
     2281 gacacgggaa agcagtccat ttcttacgac ttgcacactg agcagtgtat tgctgacaaa
     2341 agcatagcgg actgtgtgga agccctgctg ggctgctatt taaccagctg tggggagagg
     2401 gctgctcagc ttttcctctg ttcactgggg ctgaaggtgc tcccggtaat taaaaggact
     2461 gatcgggaaa aggccctgtg ccctactcgg gagaatttca acagccaaca aaagaacctt
     2521 tcagtgagct gtgctgctgc ttctgtggcc agttcacgct cttctgtatt gaaagactcg
     2581 gaatatggtt gtttgaagat tccaccaaga tgtatgtttg atcatccaga tgcagataaa
     2641 acactgaatc accttatatc ggggtttgaa aattttgaaa agaaaatcaa ctacagattc
     2701 aagaataagg cttaccttct ccaggctttt acacatgcct cctaccacta caatactatc
     2761 actgattgtt accagcgctt agaattcctg ggagatgcga ttttggacta cctcataacc
     2821 aagcaccttt atgaagaccc gcggcagcac tccccggggg tcctgacaga cctgcggtct
     2881 gccctggtca acaacaccat ctttgcatcg ctggctgtaa agtacgacta ccacaagtac
     2941 ttcaaagctg tctctcctga gctcttccat gtcattgatg actttgtgca gtttcagctt
     3001 gagaagaatg aaatgcaagg aatggattct gagcttagga gatctgagga ggatgaagag
     3061 aaagaagagg atattgaagt tccaaaggcc atgggggata tttttgagtc gcttgctggt
     3121 gccatttaca tggatagtgg gatgtcactg gagacagtct ggcaggtgta ctatcccatg
     3181 atgcggccac taatagaaaa gttttctgca aatgtacccc gttcccctgt gcgagaattg
     3241 cttgaaatgg aaccagaaac tgccaaattt agcccggctg agagaactta cgacgggaag
     3301 gtcagagtca ctgtggaagt agtaggaaag gggaaattta aaggtgttgg tcgaagttac
     3361 aggattgcca aatctgcagc agcaagaaga gccctccgaa gcctcaaagc taatcaacct
     3421 caggttccca atagctgagt cgagtctaga gggcccgttt aaacccgctg atcagcctcg
     3481 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     3541 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     3601 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     3661 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     3721 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     3781 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3841 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3901 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3961 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4021 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4081 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4141 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     4201 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4261 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4321 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4381 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4441 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     4501 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     4561 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     4621 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4681 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     4741 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     4801 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4861 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4921 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     4981 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     5041 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     5101 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     5161 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     5221 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     5281 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     5341 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     5401 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     5461 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5521 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     5581 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5641 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     5701 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5761 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5821 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5881 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5941 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6001 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6061 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6121 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6181 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6241 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6301 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6361 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6421 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6481 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6541 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6601 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6661 tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6721 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6781 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6841 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6901 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6961 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7021 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7081 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7141 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7201 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7261 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7321 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7381 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7441 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7501 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7561 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7621 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7681 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7741 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7801 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7861 ccgaaaagtg ccacctgacg tc
//
