LOCUS       sequence-307825-        7787 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Frame-shift fluorescent reporter for genomic DNA editing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7787)
  AUTHORS   Kumar A, Birnbaum MD, Moorthy BT, Singh J, Palovcak A, Patel DM, 
            Zhang F
  TITLE     Insertion/deletion-activated frame-shift fluorescence protein is a 
            sensitive reporter for genomic DNA editing.
  JOURNAL   BMC Genomics. 2019 Jul 24;20(1):609. doi: 10.1186/s12864-019-5963-z.
  PUBMED    31340764
REFERENCE   2  (bases 1 to 7787)
  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: "BMC 
            Genomics."; date: "2019-07-24"; volume: "20(1):609. doi"; pages: " 
            10.1186/s12864-019-5963-z"
COMMENT     Sequence Label: pQC-FsCFP-mCherryFP
FEATURES             Location/Qualifiers
     source          1..7787
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        2..381
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..585
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             586..761
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    824..1023
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1224..1640
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1570..1592
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1608..1624
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     enhancer        1677..1980
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1981..2184
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2181..2205
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(2366..2368,2369..3082)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2408..2429)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2669..2688)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3016..3037
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3098..3670
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3121..3672
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3265..3282)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3492..3511
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      3686..3695
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3692..4402
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3837..3855)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4105..4124
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     LTR             4566..4991
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(5094..5116)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5252..5272)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5257..5586
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5437..5572
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5608..5627
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5708..5725)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5879..6467)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5959..5978)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6638..7429,7430..7498))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7261..7280
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7499..7603)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7671..7689
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
       61 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
      121 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
      181 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      241 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
      301 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
      361 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
      421 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
      481 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
      541 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcaata aaagagccca
      601 caacccctca ctcggcgcgc cagtcttccg atagactgcg tcgcccgggt acccgtattc
      661 ccaataaagc ctcttgctgt ttgcatccga atcgtggtct cgctgttcct tgggagggtc
      721 tcctctgagt gattgactac ccacgacggg ggtctttcat ttgggggctc gtccgggatt
      781 tggagacccc tgcccaggga ccaccgaccc accaccggga ggtaagctgg ccagcaactt
      841 atctgtgtct gtccgattgt ctagtgtcta tgtttgatgt tatgcgcctg cgtctgtact
      901 agttagctaa ctagctctgt atctggcgga cccgtggtgg aactgacgag ttctgaacac
      961 ccggccgcaa ccctgggaga cgtcccaggg actttggggg ccgtttttgt ggcccgacct
     1021 gaggaaggga gtcgatgtgg aatccgaccc cgtcaggata tgtggttctg gtaggagacg
     1081 agaacctaaa acagttcccg cctccgtctg aatttttgct ttcggtttgg aaccgaagcc
     1141 gcgcgtcttg tctgctgcag cgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt
     1201 ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt ttgaccttag
     1261 gtcactggaa agatgtcgag cggatcgctc acaaccagtc ggtagatgtc aagaagagac
     1321 gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg
     1381 gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc
     1441 atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct tttgaccccc
     1501 ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttcct ccatccgccc
     1561 cgtctctccc ccttgaacct cctcgttcga ccccgcctcg atcctccctt tatccagccc
     1621 tcactccttc tctaggcgcc ggaattgaag atctggggga tcgatcctct agagtccgtt
     1681 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     1741 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     1801 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     1861 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     1921 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     1981 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     2041 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     2101 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     2161 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat
     2221 ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg cggccgggaa
     2281 cggtgcattg gaacggacct gcaggaattg atccgcggcc gccatatgtg ggtgggggga
     2341 gtttgctcct ggtgaactcg agagcgtgag caagggcgag gagctgttca ccggggtggt
     2401 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     2461 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     2521 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tggggcgtgc agtgcttcgc
     2581 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     2641 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     2701 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     2761 ggacggcaac atcctggggc acaagctgga gtacaacgcc atcagcgaca acgtctatat
     2821 caccgccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga
     2881 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     2941 cgtgctgctg cccgacaacc actacctgag cacccagtcc aagctgagca aagaccccaa
     3001 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     3061 catggacgag ctgtacaagt aaggatccgg aattccgccc ctctccctcc ccccccccta
     3121 acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt
     3181 ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga
     3241 cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg
     3301 tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt
     3361 gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat
     3421 aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg
     3481 aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg
     3541 taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt
     3601 cgaggttaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac
     3661 acgatgataa tatggccaca cgcgtgccac catggtgagc aagggcgagg aggataacat
     3721 ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca
     3781 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa
     3841 gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt
     3901 catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct
     3961 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt
     4021 gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg
     4081 cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc
     4141 ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct
     4201 gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa
     4261 gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa
     4321 cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg
     4381 catggacgag ctgtacaagt gattcgaatg atcaattaat taagaattat caagcttatc
     4441 gataccgtcg agatatcagt ggtccaggct ctagttttga ctcaacaata tcaccagctg
     4501 aagcctatag agtacgagcc atagataaaa taaaagattt tatttagtct ccagaaaaag
     4561 gggggaatga aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc
     4621 aaggcatgga aaaatacata actgagaata gagaagttca gatcaaggtc aggaacagat
     4681 ggaacagggt cgaccctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa
     4741 tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct
     4801 tctgctcccc gagctcaata aaagagccca caacccctca ctcggggcgc cagtcctccg
     4861 attgactgag tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac
     4921 ttgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg
     4981 ggtctttcat ttgggggctc gtccgggatc gggagacccc tgcccaggga ccaccgaccc
     5041 accaccggga ggtaagctgg ctgcctcgcg cgtttcggtg atgacggtga aaacctctga
     5101 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     5161 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca
     5221 cgtagcgata gcggagtgta gatccggctg tggaatgtgt gtcagttagg gtgtggaaag
     5281 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     5341 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     5401 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     5461 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     5521 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     5581 tgcaaaaagc ttactggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     5641 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt
     5701 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5761 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5821 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5881 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5941 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     6001 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     6061 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6121 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6181 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6241 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6301 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6361 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6421 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6481 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6541 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6601 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6661 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6721 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6781 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6841 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6901 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctgcaggca
     6961 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     7021 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     7081 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     7141 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     7201 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaacacggg
     7261 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     7321 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     7381 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     7441 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     7501 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7561 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     7621 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     7681 tcacgaggcc ctttcgtctt caagaattag cttggccatt gcatacgttg tatccatatc
     7741 ataatatgta catttatatt ggctcatgtc caacattacc gccatgt
//
