LOCUS       sequence-237601-        7079 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  In vivo homologous recombination substrate composed of two 
            differentially mutated GFP genes oriented as direct repeats and 
            separated by a drug selection marker .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7079)
  AUTHORS   Pierce AJ, Johnson RD, Thompson LH, Jasin M
  TITLE     XRCC3 promotes homology-directed repair of DNA damage in mammalian 
            cells.
  JOURNAL   Genes Dev. 1999 Oct 15. 13(20):2633-8.
  PUBMED    10541549
REFERENCE   2  (bases 1 to 7079)
  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"; journalName: "Genes Dev. 
            1999 Oct 15. 13(20):2633-8."
COMMENT     Sequence Label: pDRGFP
FEATURES             Location/Qualifiers
     source          1..7079
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     69..88
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             146..166
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             188..208
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      422..431
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(428..430,431..433,434..1147)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSG*G*QGNTYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     1081..1102
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1270..1391
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(2052..2651)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2135..2155)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     2632..2651
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(2670..3167)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2718..2737)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     3109..3132
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     complement(3255..3274)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3320..3375
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3374..3393)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     CDS             3739..3759
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3781..3801
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      4015..4024
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4021..4023,4024..4026,4027..4485)
                     /codon_start=1
                     /product="improved N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN155(I152L)
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMA"
     primer_bind     complement(4541..4557)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4541..4557)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4554..4576)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4565..4581
                     /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(4589..4595,4596..4613,4614..4619))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4634..4655
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4714..4909
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4760..4895
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4915..5049
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5117..5134)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5288..5876)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5368..5387)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6047..6838,6839..6907))
                     /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     6670..6689
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6908..7012)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgtgaccggc ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca
       61 gctcctgggc aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattcagatc
      121 cgccgccact atgggatcaa gatcgccaaa aaagaagaga aaggtgccga agaagcatgc
      181 agcaccacca aaaaaaaaac gaaaagtaga agacccacga ggcaacacca gcggcgtgct
      241 gagcaccccc aaggccaaga gggccaagca cccccccggc accgagaagc ccaggagcag
      301 gagccagagc gagcagcccg ccacctgccc catctgctac gccgtgatca ggcagagcag
      361 gaacctgagg aggcacctgg agctgaggca cttcgccaag cccggcgtgg atccaccggt
      421 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
      481 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggctagggat aacagggtaa
      541 tacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
      601 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
      661 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
      721 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
      781 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
      841 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
      901 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
      961 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     1021 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     1081 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     1141 caagtaaagc ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta
     1201 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     1261 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     1321 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     1381 aatgtatctt aaggcgtaaa ttgtaagcgt taatctgaag cttctgatgg aattagaact
     1441 tggcaaaaca atactgagaa tgaagtgtat gtggaacaga ggctgctgat ctcgttcttc
     1501 aggctatgaa actgacacat ttggaaacca cagtacttag aaccacaaag tgggaatcaa
     1561 gagaaaaaca atgatcccac gagagatcta tagatctata gatcatgagt gggaggaatg
     1621 agctggccct taatttggtt ttgcttgttt aaattatgat atccaactat gaaacattat
     1681 cataaagcaa tagtaaagag ccttcagtaa agagcaggca tttatctaat cccaccccac
     1741 ccccaccccc gtagctccaa tccttccatt caaaatgtag gtactctgtt ctcacccttc
     1801 ttaacaaagt atgacaggaa aaacttccat tttagtggac atctttattg tttaatagat
     1861 catcaatttc gatccgctcc tgggcaccga actgcgccgc gtgttcagca gggtcggcgt
     1921 gttcggtgtg tcccccgcgg tgggcctcgg gggcgggtgc ggggtcggcg gggccgcccc
     1981 gggtggcttc ggtcggagcc atggggtcgt gcgctccttt cggtcgggcg ctgcgggtcg
     2041 tggggcgggc gtcaggcacc gggcttgcgg gtcatgcacc aggtgcgcgg tccttcgggc
     2101 acctcgacgt cggcggtgac ggtgaagccg agccgctcgt agaaggggag gttgcggggc
     2161 gcggaggtct ccaggaaggc gggcaccccg gcgcgctcgg ccgcctccac tccggggagc
     2221 acgacggcgc tgcccagacc cttgccctgg tggtcgggcg agacgccgac ggtggccagg
     2281 aaccacgcgg gctccttggg ccggtgcggc gccaggaggc cttccatctg ttgctgcgcg
     2341 gccagccggg aaccgctcaa ctcggccatg cgcgggccga tctcggcgaa caccgccccc
     2401 gcttcgacgc tctccggcgt ggtccagacc gccaccgcgg cgccgtcgtc cgcgacccac
     2461 accttgccga tgtcgagccc gacgcgcgtg aggaagagtt cttgcagctc ggtgacccgc
     2521 tcgatgtggc ggtccgggtc gacggtgtgg cgcgtggcgg ggtagtcggc gaacgcggcg
     2581 gcgagggtgc gtacggcccg ggggacgtcg tcgcgggtgg cgaggcgcac cgtgggcttg
     2641 tactcggtca tggtggcggc tggatcggtc gaaaggcccg gagatgagga agaggagaac
     2701 agcgcggcag acgtgcgctt ttgaagcgtg cagaatgccg ggcctccgga ggaccttcgg
     2761 gcgcccgccc cgcccctgag cccgcccctg agcccgcccc cggacccacc ccttcccagc
     2821 ctctgagccc agaaagcgaa ggagcaaagc tgctattggc cgctgcccca aaggcctacc
     2881 cgcttccatt gctcagcggt gctgtccatc tgcacgagac tagtgagacg tgctacttcc
     2941 atttgtcacg tcctgcacga cgcgagctgc ggggcggggg ggaacttcct gactagggga
     3001 ggagtagaag gtggcgcgaa ggggccacca aagaacggag ccggttggcg cctaccggtg
     3061 gatgtggaat gtgtgaggcc agaggccact tgtgtagcgc caagtgccca gcggggctgc
     3121 taaagcgcat gctccagact gccttgggaa aagcgcctcc cctacccggt agaattcact
     3181 cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac
     3241 aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt
     3301 gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat
     3361 tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat
     3421 gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt
     3481 tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat
     3541 agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct
     3601 ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga
     3661 ctactcccag tcatagctgt ccctcttctc ttatggagat ccctcgacct gcagcccaag
     3721 ctttagggat caagatcgcc aaaaaagaag agaaaggtgc cgaagaagca tgcagcacca
     3781 ccaaaaaaaa aacgaaaagt agaagaccca cgaggcaaca ccagcggcgt gctgagcacc
     3841 cccaaggcca agagggccaa gcaccccccc ggcaccgaga agcccaggag caggagccag
     3901 agcgagcagc ccgccacctg ccccatctgc tacgccgtga tcaggcagag caggaacctg
     3961 aggaggcacc tggagctgag gcacttcgcc aagcccggcg tggatccacc ggtcgccacc
     4021 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     4081 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     4141 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     4201 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     4261 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     4321 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     4381 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     4441 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     4501 ggcatcaggt gaacttcaat aaaagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     4561 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     4621 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     4681 tccagtcggg aaacctgtcg tgccagcgga tccgcatctc aattagtcag caaccatagt
     4741 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4801 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     4861 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctaacttg
     4921 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     4981 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     5041 gtctggatcc gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5101 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5161 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5221 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5281 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5341 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5401 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5461 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5521 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5581 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5641 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5701 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     5761 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5821 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5881 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5941 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6001 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6061 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6121 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6181 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6241 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6301 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6361 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6421 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6481 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6541 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6601 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6661 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6721 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6781 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6841 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6901 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6961 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7021 cccgaaaagt gccacctggg tcgacattga ttattgacta gttattaata gtaatcaat
//
