LOCUS       40924_8606        6436 bp DNA     circular SYN 13-MAY-2021
DEFINITION  Homologous recombination donor for pDRR. Expresses mCherry after 
            repait by HR..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6436)
  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 6436)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6436)
  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..6436
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(87..103)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(111..127)
                     /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)."
     promoter        complement(135..165)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(180..201)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        259..455
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     polyA_signal    461..595
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(663..680)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(834..1422)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1596..2453)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2454..2558)
                     /label=AmpR promoter
     CDS             2615..3412
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
                     /translation="GSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQG
                     RPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEE
                     HQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQD
                     IALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3413..3466
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3473..4177
                     /codon_start=1
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     CDS             4183..4896
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     promoter        5024..5235
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             5298..5996
                     /codon_start=1
                     /label=TagBFP
                     /note="monomeric blue fluorescent protein"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     primer_bind     complement(6042..6061)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6107..6162
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6161..6180)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also
                     called rb-glob-pA-term-R"
ORIGIN
        1 tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga tccctcgacc
       61 tgcagcccaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
      121 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
      181 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
      241 ctgtcgtgcc agcggatccg catctcaatt agtcagcaac catagtcccg cccctaactc
      301 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
      361 ttttttttat ttatacagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
      421 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct aacttgttta ttgcagctta
      481 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
      541 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatccgctg
      601 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
      661 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
      721 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      781 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      841 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      901 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      961 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     1021 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     1081 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     1141 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     1201 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     1261 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     1321 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     1381 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     1441 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     1501 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     1561 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     1621 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     1681 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     1741 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     1801 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     1861 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     1921 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     1981 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     2041 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     2101 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     2161 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     2221 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     2281 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     2341 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     2401 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     2461 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     2521 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     2581 cctggtcgac attgattagt ccgactctca gtacggatcg gccattgaac aagatggatt
     2641 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     2701 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     2761 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     2821 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     2881 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     2941 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     3001 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     3061 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     3121 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     3181 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     3241 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     3301 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     3361 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tcgagggcag
     3421 aggaagtctt ctaacatgcg gtgacgtgga ggagaatccg ggccccatta gggtctctaa
     3481 aggagaggag gataatatgg caattatcaa ggagtttatg agatttaagg tgcacatgga
     3541 aggctccgtg aacggccacg agttcgagat tgagggagag ggcgagggac ggccctatga
     3601 gggaacacag accgccaagc tgaaggtcac caagggagga cccctgcctt tcgcatggga
     3661 catcctttca cctcagttca tgtacgggag taaagcgtat gtaaaacatc cagccgatat
     3721 tcccgactat cttaaactgt cctttcccga aggttttaag tgggagagag tgatgaattt
     3781 tgaagatggg ggcgttgtga cggttacaca ggattcttca ctccaagatg gcgagtttat
     3841 atacaaggtt aagctgcgag gcacaaattt tccttccgat ggcccagtca tgcagaaaaa
     3901 aactatgggg tgggaggcaa gctcagagcg aatgtacccc gaagatgggg ccctgaaagg
     3961 agaaattaag caaagactga aactgaagga tggcggacac tacgacgccg aggtcaagac
     4021 aacgtataag gccaaaaaac ctgtgcagct cccgggcgcc tataacgtga atatcaagct
     4081 tgacataacc tctcacaacg aagactacac tatcgtggag cagtacgagc gcgctgaggg
     4141 aaggcatagc accggaggga tggacgagtt gtataagtga gggtgagcaa gggcgaagaa
     4201 ctctttacag gcgtcgttcc catcttggtt gagcttgacg gggacgtcaa cggccataaa
     4261 ttcagcgttt ccggggaggg agagggcgat gcaacatacg ggaagctgac tttgaagttt
     4321 atctgtacga ctggaaaact gccggtgcct tggccgacgc tggtgactac tctgacatat
     4381 ggtgtgcagt gtttttccag atatcctgat catatgaagc agcatgattt cttcaagagc
     4441 gcgatgccag agggctacgt gcaggaaaga acgatcttct tcaaagacga tggtaattac
     4501 aaaactcggg cagaggtcaa attcgaggga gatacgctgg taaaccggat cgagctgaag
     4561 ggcatcgatt ttaaggaaga tggtaacatc ctgggacaca aactggagta taattataac
     4621 tcccataatg tctatattat ggctgacaag cagaagaacg ggatcaaggt gaactttaag
     4681 atccggcaca atatcgaaga tggctccgtt cagctggccg atcattatca acaaaacaca
     4741 cctattggag atgggccagt cctgttgcca gataatcact atctgtccac acagtccgcc
     4801 ttgtccaaag acccgaacga gaagagagac cacatggtgc tgctggagtt tgttacggcg
     4861 gccgggatca cgttgggcat ggatgagctt tacaagtgaa gcggccgcga ctcataatca
     4921 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     4981 acctgaaaca taaaatgaat gcaattgaat caacctctgg attgggcaga gcgcacatcg
     5041 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg
     5101 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt
     5161 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt
     5221 gccgccagaa cacagctctg gctaactaga gaacccactg tagtgaaccg tcagatcgcc
     5281 tggagacgcc atccacgatg tccgagctga ttaaggagaa catgcacatg aagctgtaca
     5341 tggagggcac cgtggacaac catcacttca agtgcacatc cgagggcgaa ggcaagccct
     5401 acgagggcac ccagaccatg agaatcaagg tggtcgaggg cggccctctc cccttcgcct
     5461 tcgacatcct ggctactagc ttcctctacg gcagcaagac cttcatcaac cacacccagg
     5521 gcatccccga cttcttcaag cagtccttcc ctgagggctt cacatgggag agagtcacca
     5581 catacgaaga cgggggcgtg ctgaccgcta cccaggacac cagcctccag gacggctgcc
     5641 tcatctacaa cgtcaagatc agaggggtga acttcacatc caacggccct gtgatgcaga
     5701 agaaaacact cggctgggag gccttcaccg agacgctgta ccccgctgac ggcggcctgg
     5761 aaggcagaaa cgacatggcc ctgaagctcg tgggcgggag ccatctgatc gcaaacatca
     5821 agaccacata tagatccaag aaacccgcta agaacctcaa gatgcctggc gtctactatg
     5881 tggactacag actggaaaga atcaaggagg ccaacaacga gacctacgtc gagcagcacg
     5941 aggtggcagt ggccagatac tgcgacctcc ctagcaaact ggggcacaag cttaattgaa
     6001 tctcatgctg gagttgccct ggctcacaaa taccactgag atctttttcc ctctgccaaa
     6061 aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt
     6121 attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg
     6181 agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc
     6241 catatgctgg ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag
     6301 ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta gatttttttt
     6361 atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta
     6421 ctagccagat ttttcc
//
