LOCUS       pLCN_DSB_Repair        8444 bp DNA     circular SYN 13-MAY-2021
DEFINITION  DSB Repair Reporter (DRR). Expresses GFP after cut of two inverted 
            ISce1 sites and repair by NHEJ..
ACCESSION   .
VERSION     .
KEYWORDS    pLCN DSB Repair Reporter (DRR).
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8444)
  AUTHORS   Arnoult N, Correia A, Ma J, Merlo A, Garcia-Gomez S, Maric M, 
            Tognetti M, Benner CW, Boulton SJ, Saghatelian A, Karlseder J
  TITLE     Regulation of DNA repair pathway choice in S and G2 phases by the 
            NHEJ inhibitor CYREN.
  JOURNAL   Nature. 2017 Sep 20;549(7673):548-552. doi: 10.1038/nature24023.
  PUBMED    28959974
REFERENCE   2  (bases 1 to 8444)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8444)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName: 
            "Nature"; date: "2017-09-20- 20"; volume: "549"; issue: "7673"; 
            pages: "548-552"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8444
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..22
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        37..67
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    75..91
                     /label=lac operator
                     /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     99..115
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        136..154
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        182..408
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             409..589
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    636..761
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1254..1487
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1672..1716
                     /label=gp41 peptide
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et 
                     al., 2013)"
     CDS             1865..1906
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
     misc_feature    1983..2099
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     enhancer        2122..2425
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2426..2629
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     primer_bind     2626..2650
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2687..3487
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
     CDS             3488..3541
                     /label=T2A
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             3584..3610
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
     CDS             3630..4346
                     /label=EGFP
                     /note="enhanced GFP"
     misc_feature    4458..5046
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             5144..5377
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5449..5583
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      5610..5745
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(5766..5784)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5794..5810)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      5952..6407
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6433..6537
                     /label=AmpR promoter
     CDS             6538..7395
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      7569..8157
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8311..8328
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
       61 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
      121 ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc tgcaagctta
      181 atgtagtctt atgcaatact cttgtagtct tgcaacatgg taacgatgag ttagcaacat
      241 gccttacaag gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag gtggtacgat
      301 cgtgccttat taggaaggca acagacgggt ctgacatgga ttggacgaac cactgaattg
      361 ccgcattgca gagatattgt atttaagtgc ctagctcgat acataaacgg gtctctctgg
      421 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
      481 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
      541 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga
      601 acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg
      661 ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga
      721 ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa
      781 ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa
      841 aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag
      901 aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat
      961 cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga
     1021 tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta
     1081 agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac
     1141 aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca
     1201 cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct
     1261 ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg
     1321 acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg
     1381 gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag
     1441 gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt
     1501 tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa
     1561 tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat
     1621 tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa
     1681 caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat
     1741 tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata
     1801 gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt
     1861 cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt
     1921 ggagagagag acagagacag atccattcga ttagtgaacg gatctcgacg gtatcggtta
     1981 acttttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata
     2041 atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaatt caaaatttta
     2101 tcgataagct tgggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     2161 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     2221 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     2281 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     2341 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     2401 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
     2461 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     2521 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     2581 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa
     2641 ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccacatgg gatcggccat
     2701 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     2761 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     2821 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     2881 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     2941 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     3001 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     3061 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     3121 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     3181 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     3241 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     3301 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     3361 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     3421 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     3481 gttcttcgag ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atccgggccc
     3541 cattagggat aacagggtaa tggcggcggc ggatccggta tgatacccat acgatgttcc
     3601 tgactatgcg attaccctgt tatccctaca tggtgagcaa gggcgaagaa ctctttacag
     3661 gcgtcgttcc catcttggtt gagcttgacg gggacgtcaa cggccataaa ttcagcgttt
     3721 ccggggaggg agagggcgat gcaacatacg ggaagctgac tttgaagttt atctgtacga
     3781 ctggaaaact gccggtgcct tggccgacgc tggtgactac tctgacatat ggtgtgcagt
     3841 gtttttccag atatcctgat catatgaagc agcatgattt cttcaagagc gcgatgccag
     3901 agggctacgt gcaggaaaga acgatcttct tcaaagacga tggtaattac aaaactcggg
     3961 cagaggtcaa attcgaggga gatacgctgg taaaccggat cgagctgaag ggcatcgatt
     4021 ttaaggaaga tggtaacatc ctgggacaca aactggagta taattataac tcccataatg
     4081 tctatattat ggctgacaag cagaagaacg ggatcaaggt gaactttaag atccggcaca
     4141 atatcgaaga tggctccgtt cagctggccg atcattatca acaaaacaca cctattggag
     4201 atgggccagt cctgttgcca gataatcact atctgtccac acagtccgcc ttgtccaaag
     4261 acccgaacga gaagagagac cacatggtgc tgctggagtt tgttacggcg gccgggatca
     4321 cgttgggcat ggatgagctt tacaagtgaa gcggccgcga ctcataatca gccataccac
     4381 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     4441 taaaatgaat gcaattgaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     4501 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     4561 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     4621 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     4681 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     4741 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     4801 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt
     4861 cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     4921 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     4981 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     5041 ccccgcttgt tgttaggcgt cgacagtact aagctttctt taagaccaat gacttacaag
     5101 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctagctcac
     5161 tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc
     5221 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     5281 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     5341 ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt
     5401 attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt
     5461 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     5521 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     5581 ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     5641 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     5701 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggg acgtacccaa
     5761 ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga
     5821 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     5881 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     5941 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     6001 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     6061 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     6121 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     6181 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     6241 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     6301 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     6361 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
     6421 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     6481 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     6541 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     6601 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     6661 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     6721 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     6781 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     6841 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     6901 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     6961 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     7021 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     7081 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     7141 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     7201 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     7261 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     7321 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     7381 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     7441 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     7501 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     7561 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     7621 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     7681 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     7741 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     7801 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     7861 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     7921 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     7981 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     8041 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     8101 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     8161 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     8221 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     8281 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     8341 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     8401 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caat
//
