LOCUS       sequence-443902-        7653 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express RNASEH1 in mammalian cell..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7653)
  AUTHORS   Chen L, Chen JY, Zhang X, Gu Y, Xiao R, Shao C, Tang P, Qian H, Luo 
            D, Li H, Zhou Y, Zhang DE, Fu XD
  TITLE     R-ChIP Using Inactive RNase H Reveals Dynamic Coupling of R-loops 
            with Transcriptional Pausing at Gene Promoters.
  JOURNAL   Mol Cell. 2017 Nov 16;68(4):745-757.e5. doi: 
            10.1016/j.molcel.2017.10.008. Epub 2017 Nov 2.
  PUBMED    29104020
REFERENCE   2  (bases 1 to 7653)
  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.molcel.2017.10.008"; journalName: "Mol Cell"; date: 
            "2017-11-16- 16"; volume: "68"; issue: "4"; pages: "745-757.e5"
COMMENT     Sequence Label: ppyCAG_RNaseH1_WT
FEATURES             Location/Qualifiers
     source          1..7653
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(435..712)
                     /label=chicken beta-actin promoter
     enhancer        714..1093
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    1107..1241
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             join(1834..1902,1903..2694)
                     /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     complement(2052..2071)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2865..3453
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3354..3373
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3607..3624
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    3692..3826
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(3832..4027)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3846..3981
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    4086..4107
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4122..4127,4128..4145,4146..4152)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4160..4176
                     /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)."
     primer_bind     4165..4187
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4184..4200
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4184..4200
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    4247..4471
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4460..4477
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(4496..5521)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(4496..5521)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     misc_feature    join(5532..5540,5541..5543,5544..6118)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5705..5724)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     5934..5951
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(6162..6203)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(6984..7004)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(7044..7063)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          join(7071..7653,1..434)
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(7125..7147)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
ORIGIN
        1 caccccccca cgcacacacc ccgcacgcag cctttgttcc cctcgcagcc ccccccgcac
       61 cgcggggcac cgcccccggc cgcgctcccc tcgcgcacac tgcggagcgc acaaagcccc
      121 gcgccgcgcc cgcagcgctc acagccgccg ggcagcgcgg agccgcacgc ggcgctcccc
      181 acgcacacac acacgcacgc accccccgag ccgctccccc cgcacaaagg gccctcccgg
      241 agcccctcaa ggctttcacg cagccacaga aaagaaacaa gccgtcatta aaccaagcgc
      301 taattacagc ccggaggaga agggccgtcc cgcccgctca cctgtgggag taacgcggtc
      361 agtcagagcc ggggcgggcg gcgcgaggcg gcggcggagc ggggcacggg gcgaaggcag
      421 cgcgcagcga ctcccgcccg ccgcgcgctt cgctttttat agggccgccg ccgccgccgc
      481 ctcgccataa aaggaaactt tcggagcgcg ccgctctgat tggctgccgc cgcacctctc
      541 cgcctcgccc cgccccgccc ctcgccccgc cccgccccgc ctggcgcgcg cccccccccc
      601 ccccccgccc ccatcgctgc acaaaataat taaaaaataa ataaatacaa aattgggggt
      661 ggggaggggg gggagatggg gagagtgaag cagaacgtgg ggctcacctc gaccatggta
      721 atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac
      781 tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc
      841 atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga
      901 cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat
      961 gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg
     1021 aactccatat atgggctatg aactaatgac cccgtaattg attactatta ataactagtc
     1081 aataatcaat gtcgacgatg atatcagatc cagacatgat aagatacatt gatgagtttg
     1141 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     1201 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taattcgatt cagcttcaga
     1261 agatggcgga gggcctccaa cacagtaatt ttcctcccga ctcttaaaat agaaaatgtc
     1321 aagtcagtta agcaggaagt gactaactga ccgcagctgg ccgtgcgaca tcctctttta
     1381 attagttgct aggcaactgc cctccagagg gcagtgtggt tttgcaagag gaagcaaaaa
     1441 gcctctccac ccaggcctgg aatgtcagtg tggttttgca agaggaagca aaaagcctct
     1501 ccacccaggc ctggaatgtt tccacccaat cattactatg acaacagctg ttttttttag
     1561 tattaagcag aggccggggg cccctggcct ccgcttactc tggagaaaaa gaagagaggc
     1621 ttccagaggc aacttgtcaa aacaggactg gcgccttgga ggcgctgtgg ggccacccaa
     1681 attgatataa ttaagcccca accgcctctt cccgcctcat ttcagcctca ccactatgtg
     1741 aatatagctg aatacacagt ttattgaata aacattaatt tccaggaaat acagtctttg
     1801 tttttccaaa gcggtcattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     1861 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     1921 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     1981 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     2041 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     2101 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     2161 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     2221 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     2281 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     2341 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     2401 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     2461 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     2521 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     2581 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     2641 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     2701 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     2761 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     2821 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     2881 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     2941 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     3001 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     3061 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     3121 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     3181 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     3241 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     3301 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     3361 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     3421 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     3481 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     3541 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     3601 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     3661 cccgcgcgtt ggccgattca ttaatgcagc ggatccagac atgataagat acattgatga
     3721 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     3781 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttagct ttttgcaaaa
     3841 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc
     3901 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
     3961 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
     4021 gagatgcgga tccgctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     4081 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     4141 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     4201 catgattacg ccaagcttgg gctgcaggtt ctttccgcct cagaagccat agagcccacc
     4261 gcatccccag catgcctgct attgtcttcc caatcctccc ccttgctgtc ctgccccacc
     4321 ccacccccca gaatagaatg acacctactc agacaatgcg atgcaatttc ctcattttat
     4381 taggaaagga cagtgggagt ggcaccttcc agggtcaagg aaggcacggg ggaggggcaa
     4441 acaacagatg gctggcaact agaaggcaca gtcgaggctg atcagcgagc tccgactatt
     4501 cctttgccct cggacgagtg ctggggcgtc ggtttccact atcggcgagt acttctacac
     4561 agccatcggt ccagacggcc gcgcttctgc gggcgatttg tgtacgcccg acagtcccgg
     4621 ctccggatcg gacgattgcg tcgcatcgac cctgcgccca agctgcatca tcgaaattgc
     4681 cgtcaaccaa gctctgatag agttggtcaa gaccaatgcg gagcatatac gcccggagcc
     4741 taggcgatcc tgcaagctcc ggatgcctcc gctcgaagta gcgcgtctgc tgctccatac
     4801 aagccaacca cggcctccag aagaagatgt tggcgacctc gtattgggaa tccccgaaca
     4861 tcgcctcgct ccagtcaatg accgctgtta tgcggccatt gtccgtcagg acattgttgg
     4921 agccgaaatc cgcgtgcacg aggtgccgga cttcggggca gtcctcggcc caaagcatca
     4981 gctcatcgag agcctgcgcg acggacgcac tgacggtgtc gtccatcaca gtttgccagt
     5041 gatacacatg gggatcagca atcgcgcata tgaaatcacg ccatgtagtg tattgaccga
     5101 ttccttgcgg tccgaatggg ccgaacccgc tcgtctggct aagatcggcc gcagcgatcg
     5161 catccatggc ctccgcgacc ggctgcagaa cagcgggcag ttcggtttca ggcaggtctt
     5221 gcaacgtgac accctgtgca cggcgggaga tgcaataggt caggctctcg ctgaattccc
     5281 caatgtcaag cacttccgga atcgggagcg cggccgatgc aaagtgccga taaacataac
     5341 gatctttgta gaaaccatcg gcgcagctat ttacccgcag gacatatcca cgccctccta
     5401 catcgaagct gaaagcacga gattcttcgc cctccgagag ctgcatcagg tcggagacgc
     5461 tgtcgaactt ttcgatcaga aacttctcga cagacgtcgc ggtgagttca ggctttttca
     5521 tctttgggtt gggttgtggc aagcttatca tcgtgttttt caaaggaaaa ccacgtcccc
     5581 gtggttcggg gggcctagac gtttttttaa cctcgactaa acacatgtaa agcatgtgca
     5641 ccgaggcccc agatcagatc ccatacaatg gggtaccttc tgggcatcct tcagcccctt
     5701 gttgaatacg cttgaggaga gccatttgac tctttccaca actatccaac tcacaacgtg
     5761 gcactggggt tgtgccgcct ttgcaggtgt atcttataca cgtggctttt ggccgcagag
     5821 gcacctgtcg ccaggtgggg ggttccgctg cctgcaaagg gtcgctacag acgttgtttg
     5881 tcttcaagaa gcttccagag gaactgcttc cttcacgaca ttcaacagac cttgcattcc
     5941 tttggcgaga ggggaaagac ccctaggaat gctcgtcaag aagacagggc caggtttccg
     6001 ggccctcaca ttgccaaaag acggcaatat ggtggaaaat aacatataga caaacgcaca
     6061 ccggccttat tccaagcggc ttcggccagt aacgttaggg gggggggagg gagagggggg
     6121 gggggcggaa ttcgatatca agcttatcga gcggccgctc acgtagaatc gagaccgagg
     6181 agagggttag ggataggctt accgtcttcc gattgtttag ctccttctct ggctaatctg
     6241 tcagcttctt cattgcctat aaatcccgaa tgaccaggaa catgcatcca ctgaatgtcc
     6301 atcccctggg taagcctctc cagtgccaca aagtcctctt tgttgatcac ctctttccct
     6361 gcacttgtct tccacccatt tttcttccaa ccttgaaccc agttagttat accatttatc
     6421 gtaaacatac tgtctgtata cagaaccagt ttattgatgt tttgagtctt tgcttgttca
     6481 atggctttgc aggctgcatg aatttccgct ctttggtttg tctgccgccc aggaagtcta
     6541 atgcctacat ttaaaggatg gcccggcccc cagtaaacgc cgattcctgc tcgcggcctt
     6601 ctacgcccat tactggagca gcagccatca gtgtagacga cgacgaagtc tcccatgtag
     6661 gaaaacgtgt ctctgctaac tggaggcgcc ggctccacgc tcggcttcat gtgctttgca
     6721 tacggctctg cgctttcatg tccatctcca tccagtggct cacggagtcg cttgctggct
     6781 ttcgcctccg attcttgtcc atgttgattt tcatgccctt ctgaaacttc cgggcttgca
     6841 gatttcctga caaaggccca ggcctcatcc tctgtggcaa acttcttaaa tctggcagca
     6901 ggaaaccggt ccacctgtgc tctgcactca ttccaggtca gaaagacccc ggtcttgcgg
     6961 cccctcctca cggcatagaa ggacaccttc ctcttcttct tgggcatggt ctcgaggaat
     7021 tctttgccaa aatgatgaga cagcacaata accagcacgt tgcccaggag ctgtaggaaa
     7081 aagaagaagg catgaacatg gttagcagag gctctagagc cgccggtcac acgccagaag
     7141 ccgaaccccg ccctgccccg tcccccccga aggcagccgt ccccccgcgg acagccccga
     7201 ggctggagag ggagaagggg acggcggcgc ggcgacgcac gaaggccctc cccgcccatt
     7261 tccttcctgc cggcgccgca ccgcttcgcc ccgcgcccgc tagagggggt gcggcggcgc
     7321 ctcccagatt tcggctccgc acagatttgg gacaaaggaa gtccctgcgc cctctcgcac
     7381 gattaccata aaaggcaatg gctgcggctc gccgcgcctc gacagccgcc ggcgctccgg
     7441 gggccgccgc gcccctcccc cgagccctcc ccggcccgag gcggccccgc cccgcccggc
     7501 acccccacct gccgccaccc cccgcccggc acggcgagcc ccgcgccacg ccccgtacgg
     7561 agccccgcac ccgaagccgg gccgtgctca gcaactcggg gaggggggtg cagggggggt
     7621 tgcagcccga ccgacgcgcc cacaccccct gct
//
