LOCUS       sequence-261132-        8425 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 8425)
  AUTHORS   Meister G, Landthaler M, Peters L, Chen PY, Urlaub H, Luhrmann R, 
            Tuschl T
  TITLE     Identification of novel argonaute-associated proteins.
  JOURNAL   Curr Biol. 2005 Dec 6;15(23):2149-55. doi: 
            10.1016/j.cub.2005.10.048. Epub 2005 Nov 10.
  PUBMED    16289642
REFERENCE   2  (bases 1 to 8425)
  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: 
            "10.1016/j.cub.2005.10.048"; journalName: "Curr Biol"; date: 
            "2005-12-6- 6"; volume: "15"; issue: "23"; pages: "2149-55"
COMMENT     Sequence Label: pFLAG/HA-MOV10
FEATURES             Location/Qualifiers
     source          1..8425
                     /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"
     regulatory      946..955
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             955..978
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             979..1005
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    1021..1045
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    3694..3705
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     protein_bind    complement(4107..4131)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4184..4225
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     intron          4306..4535
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     4518..4537
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(4607..5181,5182..5184,5185..5193)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4774..4791)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5001..5020
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5213..5812
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5213..5232)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5709..5729
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    6046..6167
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6083..6102)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(6268..6284)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6268..6284)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6281..6303)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6292..6308
                     /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(6316..6322,6323..6340,6341..6346))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6361..6382
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6499..6516)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6670..7258)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6750..6769)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7429..8220,8221..8289))
                     /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     8052..8071
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8290..8394)
                     /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 gcttggtacc
      901 gagctcggat cgatccatcg atagtgtggt ggaattctga tcgccgccgc catggactac
      961 aaggacgacg atgacaagta cccttatgac gtgcccgatt acgctttgaa ccagatatca
     1021 acaagtttgt acaaaaaagc aggctccacc atgggaacca attcagtcga catgcccagt
     1081 aagttcagct gccggcagct ccgggaggcg ggccagtgtt tcgagagttt cctggtcgtt
     1141 cggggactgg acatggagac agatcgcgag cggctgcgga ccatttataa ccgcgacttc
     1201 aagatcagct ttgggacccc cgcccctggc ttctcctcca tgctgtatgg aatgaagatt
     1261 gcaaatctgg cctacgtcac caagactcgg gtcaggttct tcagactcga ccgctgggcc
     1321 gacgtgcggt tcccagaaaa gaggagaatg aagctggggt cagatatcag caaacaccac
     1381 aagtcactgc tagccaagat cttttatgac agggctgagt atcttcatgg gaaacatggt
     1441 gtggatgtgg aagtccaggg gccccatgaa gcccgagatg ggcagctcct tatccgcctg
     1501 gatttgaacc gcaaagaggt gctgaccctg aggcttcgga atggcggaac ccagtctgtt
     1561 accctcactc acctcttccc actctgccgg acaccccagt ttgctttcta caatgaagac
     1621 caggagttgc cctgtccact gggccccggt gaatgctatg aactccatgt ccattgtaag
     1681 accagctttg tgggctactt cccagccaca gtgctctggg agctgctggg acctggggag
     1741 tcgggttcag aaggagccgg cacattctac attgcccgct tcttggctgc cgtcgcccac
     1801 agccccctgg ctgcacagct gaagcccatg actcccttca agcggacccg gatcaccgga
     1861 aaccctgtgg tgaccaatcg gatagaggaa ggagagagac ctgaccgcgc taagggctat
     1921 gacctggagt taagtatggc gctggggaca tactacccac ctccccgcct caggcagctg
     1981 ctccccatgc ttcttcaggg aacaagtatc ttcactgccc ctaaggagat cgcagagatc
     2041 aaggcccagc tggagacagc cctgaagtgg aggaactatg aggtgaagct gcggctgctg
     2101 ctgcacctgg aggaactgca gatggagcat gatatccggc actatgacct ggagtcggtg
     2161 cccatgacct gggaccctgt ggaccagaac cccaggctgc tcacgctgga ggttcctgga
     2221 gtgactgaga gccgcccctc agtgctacgg ggcgaccacc tgtttgccct tttgtcctcg
     2281 gagacacacc aggaggaccc catcacatat aagggctttg tgcacaaggt ggaattggac
     2341 cgtgtcaagc tgagcttttc catgagcctc ctgagccgct ttgtggatgg gctgaccttc
     2401 aaggtgaact ttaccttcaa ccgccagccg ctgcgagtcc agcaccgtgc cctggagctg
     2461 acagggcgct ggctgctgtg gcccatgctc tttcctgtgg cacctcggga cgtcccgctg
     2521 ctgccctcag atgtgaaact caagctgtac gaccggagtc tggagtcaaa cccagagcag
     2581 ctgcaggcca tgaggcacat tgttacgggc accacccgtc cagcccccta catcatcttt
     2641 gggcctccag gcaccggcaa gactgtcacg ttagtggagg caattaagca ggtggtgaag
     2701 cacttgccca aagcccacat cttggcctgc gctccatcca actcaggggc tgacctactc
     2761 tgtcaaaggc tccgggtcca ccttcctagc tccatctacc gcctcctggc ccccagcagg
     2821 gacatccgca tggtacctga ggacatcaag ccctgctgca actgggacgc aaagaagggg
     2881 gagtatgtat ttcccgccaa gaagaagctg caggaatacc gggtcttaat taccaccctc
     2941 atcactgccg gcaggttggt ctcggcccag tttcccattg atcacttcac acacatcttc
     3001 atcgatgagg ctggccactg catggagcct gagagtctgg tagctatagc agggctgatg
     3061 gaagtaaagg aaacaggtga tccaggaggg cagctggtgc tggcaggaga ccctcggcag
     3121 ctggggcctg tgctgcgttc cccactgacc cagaagcatg gactgggata ctcactgctg
     3181 gagcggctgc tcacctacaa ctccctgtac aagaagggcc ctgatggcta tgacccccag
     3241 ttcataacca agctgctccg caactacagg tctcatccca ccatcctgga cattcctaac
     3301 cagctctatt atgaagggga gctgcaggcc tgtgctgatg tcgtggatcg agaacgcttc
     3361 tgccgctggg cgggcctacc tcgacagggc tttcccatca tctttcacgg cgtaatgggc
     3421 aaagatgagc gtgaaggcaa cagcccatcc ttcttcaacc ctgaagaggc tgccacagtg
     3481 acttcctacc tgaagctgct cctggccccc tcctccaaga agggcaaagc tcgcctgagc
     3541 cctcgaagtg tgggcgtcat ctccccgtac cggaaacagg tggagaaaat ccgttactgc
     3601 atcaccaaac ttgacaggga gcttcgagga ctggatgaca tcaaggactt gaaggtgggt
     3661 tcagtagaag aattccaagg ccaagaacga agcgtcatcc tcatctccac cgtgcgaagc
     3721 agccagagct ttgtgcagct ggatctggac tttaatctgg gtttccttaa gaaccccaag
     3781 aggttcaatg tagctgtgac ccgggccaag gccctgctca tcatcgtggg gaaccccctt
     3841 ctcctgggcc atgaccctga ctggaaagta ttcctggagt tctgtaaaga aaacggaggg
     3901 tataccgggt gtcccttccc tgccaaactg gacctgcaac agggacagaa tttactgcaa
     3961 ggtctgagca agctcagccc ctctacctca gggccccaca gccatgacta cctcccccag
     4021 gagcgggagg gtgaaggggg cctgtctctg caagtggagc cagagtggag gaatgagctc
     4081 tgagcggccg cactcgagat atctagaccc agctttcttg tacaaagtgg ttgatatcca
     4141 gcacagtggc ggccgctcaa gtctagaggg cccgcggttc gaaggtaagc ctatccctaa
     4201 ccctctcctc ggtctcgatt ctacgcgtac ctgatccgcg gccgcataga taactgatcc
     4261 agtgtgctgg aattaattcg ctgtctgcga gggccagctg ttggggtgag tactccctct
     4321 caaaagcggg catgacttct gcgctaagat tgtcagtttc caaaaacgag gaggatttga
     4381 tattcacctg gcccgcggtg atgcctttga gggtggccgc gtccatctgg tcagaaaaga
     4441 caatcttttt gttgtcaagc ttgaggtgtg gcaggcttga gatctggcca tacacttgag
     4501 tgacaatgac atccactttg cctttctctc cacaggtgtc cactcccagg tccaactgca
     4561 ggtcgagcat gcatctaggg cggccaattc cgcccctctc cccccccccc tctccctccc
     4621 ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata
     4681 tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg
     4741 tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt
     4801 tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag
     4861 cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc
     4921 cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga
     4981 tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg
     5041 cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat
     5101 gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc
     5161 tttgaaaaac acgatgataa gcttgccaca acccacaagg agacgacctt ccatgaccga
     5221 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg tacgcaccct
     5281 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg accgccacat
     5341 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     5401 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     5461 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     5521 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     5581 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     5641 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     5701 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     5761 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gacgcccgcc
     5821 ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catggctccg accgaagcca
     5881 cccggggcgg ccccgccgac cccgcacccg cccccgaggc ccaccgactc tagataactg
     5941 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     6001 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     6061 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     6121 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg ctcgagtgca
     6181 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     6241 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     6301 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     6361 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     6421 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6481 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6541 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6601 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6661 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6721 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6781 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6841 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6901 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6961 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     7021 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     7081 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     7141 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     7201 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     7261 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     7321 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     7381 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     7441 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7501 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7561 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7621 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7681 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7741 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7801 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7861 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7921 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7981 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     8041 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     8101 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     8161 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     8221 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     8281 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     8341 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     8401 tccccgaaaa gtgccacctg acgtc
//
