LOCUS       sequence-364410-        6436 bp    DNA     circular SYN 15-DEC-2025
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)
  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"; journalName: 
            "Nature"; date: "2017-09-20- 20"; volume: "549"; issue: "7673"; 
            pages: "548-552"
COMMENT     Sequence Label: pCAGGS DRR mCherry Donor EF1a BFP
FEATURES             Location/Qualifiers
     source          1..6436
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     133..152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     386..403
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    471..605
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(495..514)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     549..568
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(611..806)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      625..760
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    865..886
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(901..906,907..924,925..931)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    939..955
                     /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     944..966
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     963..979
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     963..979
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1322..1341
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    1340..1395
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     1441..1460
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(1503..2204)
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     promoter        complement(2267..2478)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             complement(join(2606..3316,3317..3319))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             complement(4036..4089)
                     /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             complement(4090..4887)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="GSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQG
                     RPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEE
                     HQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQD
                     IALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4198..4217)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4808..4827
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        4944..5048
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5049..5117,5118..5909)
                     /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(5267..5286)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6080..6436,1..232)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
       61 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      241 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      301 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
      361 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
      421 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagcg gatccagaca
      481 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
      541 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
      601 aagttagctt tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata
      661 gctcagaggc cgaggcggcc tcggcctctg tataaataaa aaaaattagt cagccatggg
      721 gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga
      781 ctatggttgc tgactaattg agatgcggat ccgctggcac gacaggtttc ccgactggaa
      841 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
      901 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
      961 cacaggaaac agctatgacc atgattacgc caagcttggg ctgcaggtcg agggatctcc
     1021 ataagagaag agggacagct atgactggga gtagtcagga gaggaggaaa aatctggcta
     1081 gtaaaacatg taaggaaaat tttagggatg ttaaagaaaa aaataacaca aaacaaaata
     1141 taaaaaaaat ctaacctcaa gtcaaggctt ttctatggaa taaggaatgg acagcagggg
     1201 gctgtttcat atactgatga cctctttata gccaaccttt gttcatggca gccagcatat
     1261 gggcatatgt tgccaaactc taaaccaaat actcattctg atgttttaaa tgatttgccc
     1321 tcccatatgt ccttccgagt gagagacaca aaaaattcca acacactatt gcaatgaaaa
     1381 taaatttcct ttattagcca gaagtcagat gctcaagggg cttcatgatg tccccataat
     1441 ttttggcaga gggaaaaaga tctcagtggt atttgtgagc cagggcaact ccagcatgag
     1501 attcaattaa gcttgtgccc cagtttgcta gggaggtcgc agtatctggc cactgccacc
     1561 tcgtgctgct cgacgtaggt ctcgttgttg gcctccttga ttctttccag tctgtagtcc
     1621 acatagtaga cgccaggcat cttgaggttc ttagcgggtt tcttggatct atatgtggtc
     1681 ttgatgtttg cgatcagatg gctcccgccc acgagcttca gggccatgtc gtttctgcct
     1741 tccaggccgc cgtcagcggg gtacagcgtc tcggtgaagg cctcccagcc gagtgttttc
     1801 ttctgcatca cagggccgtt ggatgtgaag ttcacccctc tgatcttgac gttgtagatg
     1861 aggcagccgt cctggaggct ggtgtcctgg gtagcggtca gcacgccccc gtcttcgtat
     1921 gtggtgactc tctcccatgt gaagccctca gggaaggact gcttgaagaa gtcggggatg
     1981 ccctgggtgt ggttgatgaa ggtcttgctg ccgtagagga agctagtagc caggatgtcg
     2041 aaggcgaagg ggagagggcc gccctcgacc accttgattc tcatggtctg ggtgccctcg
     2101 tagggcttgc cttcgccctc ggatgtgcac ttgaagtgat ggttgtccac ggtgccctcc
     2161 atgtacagct tcatgtgcat gttctcctta atcagctcgg acatcgtgga tggcgtctcc
     2221 aggcgatctg acggttcact acagtgggtt ctctagttag ccagagctgt gttctggcgg
     2281 caaacccgtt gcgaaaaaga acgttcacgg cgactactgc acttatatac ggttctcccc
     2341 caccctcggg aaaaaggcgg agccagtaca cgacatcact ttcccagttt accccgcgcc
     2401 accttctcta ggcaccggtt caattgccga cccctccccc caacttctcg gggactgtgg
     2461 gcgatgtgcg ctctgcccaa tccagaggtt gattcaattg cattcatttt atgtttcagg
     2521 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg
     2581 ctgattatga gtcgcggccg cttcacttgt aaagctcatc catgcccaac gtgatcccgg
     2641 ccgccgtaac aaactccagc agcaccatgt ggtctctctt ctcgttcggg tctttggaca
     2701 aggcggactg tgtggacaga tagtgattat ctggcaacag gactggccca tctccaatag
     2761 gtgtgttttg ttgataatga tcggccagct gaacggagcc atcttcgata ttgtgccgga
     2821 tcttaaagtt caccttgatc ccgttcttct gcttgtcagc cataatatag acattatggg
     2881 agttataatt atactccagt ttgtgtccca ggatgttacc atcttcctta aaatcgatgc
     2941 ccttcagctc gatccggttt accagcgtat ctccctcgaa tttgacctct gcccgagttt
     3001 tgtaattacc atcgtctttg aagaagatcg ttctttcctg cacgtagccc tctggcatcg
     3061 cgctcttgaa gaaatcatgc tgcttcatat gatcaggata tctggaaaaa cactgcacac
     3121 catatgtcag agtagtcacc agcgtcggcc aaggcaccgg cagttttcca gtcgtacaga
     3181 taaacttcaa agtcagcttc ccgtatgttg catcgccctc tccctccccg gaaacgctga
     3241 atttatggcc gttgacgtcc ccgtcaagct caaccaagat gggaacgacg cctgtaaaga
     3301 gttcttcgcc cttgctcacc ctcacttata caactcgtcc atccctccgg tgctatgcct
     3361 tccctcagcg cgctcgtact gctccacgat agtgtagtct tcgttgtgag aggttatgtc
     3421 aagcttgata ttcacgttat aggcgcccgg gagctgcaca ggttttttgg ccttatacgt
     3481 tgtcttgacc tcggcgtcgt agtgtccgcc atccttcagt ttcagtcttt gcttaatttc
     3541 tcctttcagg gccccatctt cggggtacat tcgctctgag cttgcctccc accccatagt
     3601 ttttttctgc atgactgggc catcggaagg aaaatttgtg cctcgcagct taaccttgta
     3661 tataaactcg ccatcttgga gtgaagaatc ctgtgtaacc gtcacaacgc ccccatcttc
     3721 aaaattcatc actctctccc acttaaaacc ttcgggaaag gacagtttaa gatagtcggg
     3781 aatatcggct ggatgtttta catacgcttt actcccgtac atgaactgag gtgaaaggat
     3841 gtcccatgcg aaaggcaggg gtcctccctt ggtgaccttc agcttggcgg tctgtgttcc
     3901 ctcatagggc cgtccctcgc cctctccctc aatctcgaac tcgtggccgt tcacggagcc
     3961 ttccatgtgc accttaaatc tcataaactc cttgataatt gccatattat cctcctctcc
     4021 tttagagacc ctaatggggc ccggattctc ctccacgtca ccgcatgtta gaagacttcc
     4081 tctgccctcg aagaactcgt caagaaggcg atagaaggcg atgcgctgcg aatcgggagc
     4141 ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct cttcagcaat
     4201 atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc ggccacagtc
     4261 gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg catcgccatg
     4321 ggtcacgacg agatcctcgc cgtcgggcat gcgcgccttg agcctggcga acagttcggc
     4381 tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac cggcttccat
     4441 ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg
     4501 atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct cggcaggagc
     4561 aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc agtcccttcc
     4621 cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga
     4681 tagccgcgct gcctcgtcct gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa
     4741 aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt
     4801 ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag aacctgcgtg
     4861 caatccatct tgttcaatgg ccgatccgta ctgagagtcg gactaatcaa tgtcgaccag
     4921 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4981 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     5041 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     5101 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     5161 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     5221 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     5281 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     5341 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     5401 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     5461 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     5521 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     5581 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     5641 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     5701 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     5761 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     5821 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     5881 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     5941 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     6001 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     6061 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     6121 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     6181 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     6241 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     6301 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     6361 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     6421 ccagcttgga gcgaac
//
