LOCUS       sequence-154201-        6897 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a catalytically inactive form of human PMR1 endonuclease 
            with N-terminal FLAG and C-terminal biotinylation tags.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6897)
  AUTHORS   Gu SQ, Bakthavachalu B, Han J, Patil DP, Otsuka Y, Guda C, 
            Schoenberg DR
  TITLE     Identification of the human PMR1 mRNA endonuclease as an 
            alternatively processed product of the gene for peroxidasin-like 
            protein.
  JOURNAL   RNA. 2012 Jun;18(6):1186-96. doi: 10.1261/rna.031369.111. Epub 2012 
            Apr 27.
  PUBMED    22543864
REFERENCE   2  (bases 1 to 6897)
  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.031369.111"; journalName: "RNA"; date: "2012-06"; 
            volume: "18"; issue: "6"; pages: "1186-96"
COMMENT     Sequence Label: pcDNA4/TO/FLAG-hPMR1(2H)-Tev-Bio
FEATURES             Location/Qualifiers
     source          1..6897
                     /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"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             993..1016
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      1530..1539
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2589..2609
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(2908..2925)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2914..3138
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3184..3612
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3271..3290)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3481..3502
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3621..3641)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3626..3955
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3806..3941
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3868..3887
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4003..4050
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4069..4443
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4573..4694
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4610..4629)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4664..4683
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4743..4759)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4743..4759)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4756..4778)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4767..4783
                     /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(4791..4797,4798..4815,4816..4821))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4836..4857
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4974..4991)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5145..5730)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5225..5244)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5901..6692,6693..6761))
                     /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     6524..6543
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6762..6866)
                     /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 ttttggaacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtacca tggactacaa ggacgacgat gacaagatgg
     1021 ccatggaggc catgctcatg cactggggct ggtttctaga gcacgacttg gaccacacag
     1081 tgcctgcgct gagcacagcc cgcttctcgg atgggcggcc gtgcagctcc gtctgcacca
     1141 acgaccctcc ttgtttcccc atgaacaccc ggcacgccga cccccggggc acccacgcgc
     1201 cctgcatgct cttcgcgcgc tccagccccg cgtgtgccag cggccgtccc tctgcgacgg
     1261 tggattcagt ctatgcacga gagcagatca accagcaaac agcctacatc gatggctcca
     1321 acgtttacgg gagctcggag cgggaatccc aggctctcag agacccttcg gtgcctcggg
     1381 gtctcctgaa gacaggcttt ccttggcctc cctccggaaa gcccttattg cccttttcta
     1441 caggcccacc caccgagtgc gcgcgacagg agcaggagag cccctgtttc ctggccgggg
     1501 accaccgggc caacgagcat ctggctctgg ccgccatggc taccctgtgg ttccgggaac
     1561 acaacaggat ggccacggag ctgtccgccc tgaaccccca ctgggaggga aacacggttt
     1621 accaggaagc caggaagatc gtgggcgcgg agctgcagca catcacctac agccactggc
     1681 tgcctaaggt cctgggggac cctggcacta ggatgctgag gggttaccga ggctacaacc
     1741 ccaacgtgaa tgcaggcatc attaactctt ttgctactgc agcctttaga tttggcgcca
     1801 cattaatcaa tcctattctt taccgactga atgccacctt aggtgaaatt tccgaaggcc
     1861 accttccgtt ccataaagcg ctcttttcac cgtccagaat aatcaaggaa ggtgggatag
     1921 acccggttct ccgggggctg tttggcgtgg ctgctaaatg gcgggcaccc tcctaccttc
     1981 tcagtcctga gctgacccag aggctcttct ccgcggctta ttctgcggcc gtggattcgg
     2041 ctgccaccat cattcaaagg ggtagagacc acgggatccc accatatgtt gacttcagag
     2101 ttttctgtaa tttgacttca gttaagaact ttgaggatct tcaaaatgaa attaaagatt
     2161 cagagattag acaaaaactg agaaagttgt acggctctcc aggtgacatt gacctctggc
     2221 ccgcccttat ggttgaagac ctgattcctg gtacaagagt gggaccaaca cttatgtgcc
     2281 tgtttgttac ccagtttcag cggctaagag atggagatag gttctggtat gaaaaccctg
     2341 gagtatttac cccggcacaa ctcactcagc tgaagcaggc gtccctgagc cgggtgcttt
     2401 gtgacaatgg tgacagcatt cagcaagtgc aggctgatgt ctttgtaaag gcagaatacc
     2461 cacaggatta cctgaactgc agcgagatcc cgaaggtgga cctgcgagtg tggcaagact
     2521 gctgtgcaga taaacaagct ggaggcacgc ctgaggcagg cagggtgtac agatgtgcgg
     2581 cggccgcgga gaatctgtac tttcaatcag agttaaaaac cgccgcacta gctcaacatg
     2641 acgaggctgc tggaaaggcc ggtgaaggtg aaatccctgc ccctcttgct ggtaccgttt
     2701 ctaagatact ggtaaaagaa ggtgacactg ttaaagctgg tcaaacagtt ctggtgctgg
     2761 aggctatgaa aatggagaca gaaattaacg ctcctactga cggaaaagtt gaaaaggtgt
     2821 tagttaagga aagagatgct gttcaaggtg gtcaaggtct aatcaagatc ggcgtttaat
     2881 gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     2941 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     3001 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     3061 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     3121 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     3181 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     3241 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     3301 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     3361 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     3421 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     3481 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     3541 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     3601 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3661 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3721 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3781 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3841 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     3901 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     3961 cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg
     4021 gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat ggccaagttg
     4081 accagtgccg ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga gttctggacc
     4141 gaccggctcg ggttctcccg ggacttcgtg gaggacgact tcgccggtgt ggtccgggac
     4201 gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg tgccggacaa caccctggcc
     4261 tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg
     4321 aacttccggg acgcctccgg gccggccatg accgagatcg gcgagcagcc gtgggggcgg
     4381 gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga ggagcaggac
     4441 tgacacgtgc tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4501 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     4561 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     4621 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     4681 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     4741 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     4801 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     4861 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     4921 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     4981 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5041 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5101 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5161 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5221 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5281 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5341 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5401 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5461 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5521 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5581 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5641 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag
     5701 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5761 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5821 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5881 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5941 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     6001 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     6061 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6121 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6181 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6241 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6301 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6361 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6421 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6481 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6541 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6601 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6661 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6721 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6781 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6841 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
