LOCUS       sequence-9410-ex        7860 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 7860)
  AUTHORS   Landthaler M, Gaidatzis D, Rothballer A, Chen PY, Soll SJ, Dinic L, 
            Ojo T, Hafner M, Zavolan M, Tuschl T
  TITLE     Molecular characterization of human Argonaute-containing 
            ribonucleoprotein complexes and their bound target mRNAs.
  JOURNAL   RNA. 2008 Dec;14(12):2580-96. doi: 10.1261/rna.1351608. Epub 2008 
            Oct 31.
  PUBMED    18978028
REFERENCE   2  (bases 1 to 7860)
  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: 
            "10.1261/rna.1351608"; journalName: "RNA"; date: "2008-12"; volume: 
            "14"; issue: "12"; pages: "2580-96"
COMMENT     Sequence Label: pFRT/FLAG/HA-DEST EIF2C1
FEATURES             Location/Qualifiers
     source          1..7860
                     /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    complement(3656..3680)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3733..3774
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3812..3829)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3818..4042
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4175..4194)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4326..4373
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             4381..5403
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4381..5403
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    5533..5654
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5570..5589)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5624..5643
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5703..5719)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5703..5719)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5716..5738)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5727..5743
                     /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(5751..5757,5758..5775,5776..5781))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5796..5817
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5934..5951)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6105..6693)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6185..6204)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6864..7655,7656..7724))
                     /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     7487..7506
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7725..7829)
                     /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 gagctcggat cgatccatcg atagtgtggt ggaattctga tcgccgccgc catggactac
      961 aaggacgacg atgacaagta cccttatgac gtgcccgatt acgctttgaa ccagatatca
     1021 acaagtttgt acaaaaaagc aggctccacc atggaagcgg gaccctcggg agcagctgcg
     1081 ggcgcttacc tgccccccct gcagcaggtg ttccaggcac ctcgccggcc tggcattggc
     1141 actgtgggga aaccaatcaa gctcctggcc aattactttg aggtggacat ccctaagatc
     1201 gacgtgtacc actacgaggt ggacatcaag ccggataagt gtccccgtag agtcaaccgg
     1261 gaagtggtgg aatacatggt ccagcatttc aagcctcaga tctttggtga tcgcaagcct
     1321 gtgtatgatg gaaagaagaa catttacact gtcacagcac tgcccattgg caacgaacgg
     1381 gtcgactttg aggtgacaat ccctggggaa gggaaggatc gaatctttaa ggtctccatc
     1441 aagtggctag ccattgtgag ctggcgaatg ctgcatgagg ccctggtcag cggccagatc
     1501 cctgttccct tggagtctgt gcaagccctg gatgtggcca tgaggcacct ggcatccatg
     1561 aggtacaccc ctgtgggccg ctccttcttc tcaccgcctg agggctacta ccacccgctg
     1621 gggggtgggc gcgaggtctg gttcggcttt caccagtctg tgcgccctgc catgtggaag
     1681 atgatgctca acattgatgt ctcagccact gccttttata aggcacagcc agtgattgag
     1741 ttcatgtgtg aggtgctgga catcaggaac atagatgagc agcccaagcc cctcacggac
     1801 tctcagcgcg ttcgcttcac caaggagatc aagggcctga aggtggaagt cacccactgt
     1861 ggacagatga agaggaagta ccgcgtgtgt aatgttaccc gtcgccctgc tagccatcag
     1921 acattcccct tacagctgga gagtggacag actgtggagt gcacagtggc acagtatttc
     1981 aagcagaaat ataaccttca gctcaagtat ccccatctgc cctgcctaca agttggccag
     2041 gaacaaaagc atacctacct tcccctagag gtctgtaaca ttgtggctgg gcagcgctgt
     2101 attaaaaagc tgaccgacaa ccagacctcg accatgataa aggccacagc tagatccgct
     2161 ccagacagac aggaggagat cagtcgcctg atgaagaatg ccagctacaa cttagatccc
     2221 tacatccagg aatttgggat caaagtgaag gatgacatga cggaggtgac agggcgagtg
     2281 ctgccggcgc ccatcttgca gtacggcggc cggaaccggg ccattgccac acccaatcag
     2341 ggtgtctggg acatgcgggg gaaacagttc tacaatggga ttgagatcaa agtctgggcc
     2401 atcgcctgct tcgcacccca aaaacagtgt cgagaagagg tgctcaagaa cttcacagac
     2461 cagctgcgga agatttccaa ggatgcgggg atgcctatcc agggtcaacc ttgtttctgc
     2521 aaatatgcac agggggcaga cagcgtggag cctatgttcc ggcatctcaa gaacacctac
     2581 tcagggctgc agctcattat tgtcatcctg ccagggaaga cgccggtgta tgctgaggtg
     2641 aaacgtgtcg gagatacact cttgggaatg gctacgcagt gtgtgcaggt gaagaacgtg
     2701 gtcaagacct cacctcagac tctgtccaac ctctgcctca agatcaatgt caaacttggt
     2761 ggcattaaca acatcctagt cccacaccag cgctctgccg tttttcaaca gccagtgata
     2821 ttcctgggag cagatgttac acacccccca gcaggggatg ggaaaaaacc ttctatcaca
     2881 gcagtggtag gcagtatgga tgcccacccc agccgatact gtgctactgt gcgggtacag
     2941 cgaccacggc aagagatcat tgaagacttg tcctacatgg tgcgtgagct cctcatccaa
     3001 ttctacaagt ccacccgttt caagcctacc cgcatcatct tctaccgaga tggggtgcct
     3061 gaaggccagc taccccagat actccactat gagctactgg ccattcgtga tgcctgcatc
     3121 aaactggaaa aggactacca gcctgggatc acttatattg tggtgcagaa acgccatcac
     3181 acccgccttt tctgtgctga caagaatgag cgaattggga agagtggtaa catcccagct
     3241 gggaccacag tggacaccaa catcacccac ccatttgagt ttgacttcta tctgtgcagc
     3301 cacgcaggca tccagggcac cagccgacca tcccattact atgttctttg ggatgacaac
     3361 cgtttcacag cagatgagct ccagatcctg acgtaccagc tgtgccacac ttacgtacga
     3421 tgcacacgct ctgtctctat cccagcacct gcctactatg cccgcctggt ggctttccgg
     3481 gcacgatacc acctggtgga caaggagcat gacagtggag aggggagcca catatcgggg
     3541 cagagcaatg ggcgggaccc ccaggccctg gccaaagccg tgcaggttca ccaggatact
     3601 ctgcgcacca tgtacttcgc ttgacggaat tcgcggccgc actcgagata tctagaccca
     3661 gctttcttgt acaaagtggt tgatatccag cacagtggcg gccgctcaag tctagagggc
     3721 ccgcggttcg aaggtaagcc tatccctaac cctctcctcg gtctcgattc tacgctcgag
     3781 tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     3841 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     3901 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     3961 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     4021 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     4081 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4141 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     4201 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     4261 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     4321 acctagaagt tcctattccg aagttcctat tctctagaaa gtataggaac ttccttggcc
     4381 aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc
     4441 gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag cttcgatgta
     4501 ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta caaagatcgt
     4561 tatgtttatc ggcactttgc atcggccgcg ctcccgattc cggaagtgct tgacattggg
     4621 gaattcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa
     4681 gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc catggatgcg
     4741 atcgctgcgg ccgatcttag ccagacgagc gggttcggcc cattcggacc gcaaggaatc
     4801 ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca tgtgtatcac
     4861 tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg
     4921 atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc
     4981 aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag cgaggcgatg
     5041 ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg gttggcttgt
     5101 atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg atcgccgcgg
     5161 ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt ggttgacggc
     5221 aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg atccggagcc
     5281 gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt
     5341 gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa
     5401 tagcacgtac tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     5461 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     5521 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     5581 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     5641 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     5701 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     5761 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     5821 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     5881 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5941 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     6001 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     6061 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     6121 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     6181 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     6241 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     6301 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6361 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6421 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     6481 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     6541 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     6601 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     6661 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6721 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6781 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6841 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6901 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6961 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     7021 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     7081 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     7141 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     7201 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     7261 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     7321 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     7381 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     7441 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     7501 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     7561 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     7621 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     7681 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     7741 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     7801 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
