LOCUS       sequence-9413-ex        7872 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 7872)
  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 7872)
  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 EIF2C4
FEATURES             Location/Qualifiers
     source          1..7872
                     /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(3668..3692)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3745..3786
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3824..3841)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3830..4054
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4187..4206)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4338..4385
                     /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             4393..5415
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4393..5415
                     /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    5545..5666
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5582..5601)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5636..5655
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5715..5731)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5715..5731)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5728..5750)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5739..5755
                     /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(5763..5769,5770..5787,5788..5793))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5808..5829
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5946..5963)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6117..6705)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6197..6216)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6876..7667,7668..7736))
                     /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     7499..7518
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7737..7841)
                     /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 atggaggcgc tgggacccgg acctccggct
     1081 agcctgtttc agccacctcg tcgtcctggc cttggaactg ttggaaaacc aattcgactg
     1141 ttagccaatc attttcaggt tcagattcct aaaatagatg tgtatcacta tgatgtggat
     1201 attaagcctg aaaaacggcc tcgtagagtc aacagggagg tagtagatac aatggtgcgg
     1261 cacttcaaga tgcaaatatt tggtgatcgg cagcctgggt atgatggcaa aagaaacatg
     1321 tacacagcac atccactacc aattggacgg gatagggttg atatggaggt gactcttcca
     1381 ggcgagggta aagaccaaac atttaaagtg tctgttcagt gggtgtcagt tgtgagcctt
     1441 cagttgcttt tagaagcttt ggctgggcac ttgaatgaag tcccagatga ctcagtacaa
     1501 gcacttgatg ttatcacaag acaccttccc tccatgaggt acaccccagt gggccgttcc
     1561 tttttctcac ccccggaagg ttactaccac cctctgggag ggggcaggga ggtctggttt
     1621 ggttttcatc agtctgtgag acctgccatg tggaatatga tgctcaacat tgatgtatct
     1681 gcaactgctt tctaccgggc tcagcctatc attgagttca tgtgtgaggt tttagacatt
     1741 cagaacatca atgaacagac caaacctcta acagactccc agcgtgtcaa atttaccaaa
     1801 gaaatcagag gtctcaaagt tgaggtgacc cactgtggac agatgaaacg aaaataccga
     1861 gtttgtaatg tgactagacg gccagccagt catcaaactt ttcctttgca gctagaaaac
     1921 ggtcaagcta tggaatgtac agtagctcaa tattttaagc aaaagtatag tctgcaactg
     1981 aaataccccc atcttccctg tctccaagtg ggacaagaac aaaagcatac atacttgcca
     2041 ctcgaggtct gtaatatagt ggcaggacag cgatgtatca agaagctcac agacaatcag
     2101 acttccacaa tgatcaaagc tacagcaaga tctgctcctg acagacagga agagatcagt
     2161 agactggtga agagcaacag tatggtgggt ggacctgatc cataccttaa agaatttggt
     2221 attgttgtcc acaatgaaat gacagagctc acaggcaggg tacttccagc accaatgctg
     2281 caatatggag gccggaataa aacagtagcc acacccaacc agggtgtctg ggacatgcga
     2341 ggaaagcagt tttatgctgg cattgaaatt aaagtttggg cagttgcttg ttttgcacct
     2401 cagaaacaat gtagggaaga tttactaaag agtttcactg accagctgcg taaaatctct
     2461 aaggatgcag gaatgcccat ccagggtcag ccatgtttct gcaagtatgc acaaggtgca
     2521 gacagtgtgg agcctatgtt taaacatctg aaaatgactt atgtgggcct acagctaata
     2581 gtggttatcc tgcctggaaa gacaccagta tatgcggagg tgaaacgtgt tggagatacc
     2641 cttctaggta tggccacaca gtgtgtccag gtaaaaaatg tagtgaagac ctcacctcaa
     2701 accctttcca atctttgcct gaagataaat gcaaaacttg gaggaattaa caatgtgctt
     2761 gtgcctcatc aaaggccctc ggtgttccag cagcctgtca tcttcctggg agcggatgtc
     2821 acacaccccc cagcagggga tgggaagaaa ccttccattg ctgctgtggt tggcagtatg
     2881 gatggccacc ccagccggta ctgtgccacc gttcgggtgc agacttcccg gcaggagatc
     2941 tcccaagagc tcctctacag tcaagaggtc atccaggacc tgactaacat ggttcgagag
     3001 ctgctgattc agttctacaa atccacacgc ttcaaaccca ctcggatcat ctattaccgt
     3061 ggaggggtat ctgagggaca aatgaaacag gtagcttggc cagaactaat agcaattcga
     3121 aaggcatgta ttagcttgga agaagattac cggccaggaa taacttatat tgtggtgcaa
     3181 aaaagacatc acacacgact cttctgtgca gataaaacag aaagggtagg gaaaagtggc
     3241 aatgtaccag caggcactac agtggatagt accatcacac atccatctga gtttgacttt
     3301 tacctctgta gtcatgcagg aattcaggga accagccgtc cctcacatta ccaggtcttg
     3361 tgggatgaca actgcttcac tgcagatgaa ctccagctac tgacttacca gctgtgtcac
     3421 acctatgtga ggtgcactcg ctcagtctct attccagccc ctgcatatta tgcccggctt
     3481 gtagcattta gggcaaggta tcatctggtg gataaagatc atgacagtgc ggaaggcagt
     3541 catgtgtcag gacagagcaa cggccgggat cctcaggcct tggctaaggc tgtgcaaatc
     3601 caccatgata cccagcacac gatgtatttt gcctgagaga attcgcggcc gcactcgaga
     3661 tatctagacc cagctttctt gtacaaagtg gttgatatcc agcacagtgg cggccgctca
     3721 agtctagagg gcccgcggtt cgaaggtaag cctatcccta accctctcct cggtctcgat
     3781 tctacgctcg agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     3841 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     3901 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     3961 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     4021 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     4081 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4141 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     4201 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     4261 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     4321 gatggttcac gtacctagaa gttcctattc cgaagttcct attctctaga aagtatagga
     4381 acttccttgg ccaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa
     4441 aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc
     4501 agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc
     4561 tacaaagatc gttatgttta tcggcacttt gcatcggccg cgctcccgat tccggaagtg
     4621 cttgacattg gggaattcag cgagagcctg acctattgca tctcccgccg tgcacagggt
     4681 gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag
     4741 gccatggatg cgatcgctgc ggccgatctt agccagacga gcgggttcgg cccattcgga
     4801 ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc
     4861 catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct
     4921 ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg
     4981 gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg
     5041 agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg
     5101 tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca
     5161 ggatcgccgc ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc
     5221 ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc
     5281 cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg
     5341 accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg
     5401 agggcaaagg aatagcacgt actacgagat ttcgattcca ccgccgcctt ctatgaaagg
     5461 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     5521 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     5581 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     5641 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     5701 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5761 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     5821 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5881 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5941 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     6001 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     6061 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     6121 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     6181 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     6241 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     6301 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6361 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6421 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6481 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6541 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6601 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6661 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6721 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6781 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6841 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6901 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6961 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     7021 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     7081 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     7141 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     7201 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     7261 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     7321 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     7381 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     7441 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     7501 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7561 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7621 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7681 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7741 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7801 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7861 ccacctgacg tc
//
