LOCUS       sequence-207706-        4856 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses As crRNA and mCherry in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4856)
  AUTHORS   Toth E, Czene BC, Kulcsar PI, Krausz SL, Talas A, Nyeste A, Varga E,
            Huszar K, Weinhardt N, Ligeti Z, Borsy AE, Fodor E, Welker E
  TITLE     Mb- and FnCpf1 nucleases are active in mammalian cells: activities 
            and PAM preferences of four wild-type Cpf1 nucleases and of their 
            altered PAM specificity variants.
  JOURNAL   Nucleic Acids Res. 2018 Sep 20. pii: 5103950. doi: 
            10.1093/nar/gky815.
  PUBMED    30239882
REFERENCE   2  (bases 1 to 4856)
  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: "Nucleic 
            Acids Res. 2018 Sep 20. pii: 5103950. doi: 10.1093/nar/gky815."
COMMENT     Sequence Label: pTE4560
FEATURES             Location/Qualifiers
     source          1..4856
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          134..361
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      373..382
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             379..1089
                     /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(524..542)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    1218..1339
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1255..1274)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1309..1328
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1469..1488
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1602..1706
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1708..2065
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1708..1899
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1731..1751)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1916..2051
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2100..2894
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2100..2894
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2154..2173)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2764..2783
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3082..3101)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3126..3173
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3502..4090
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3991..4010
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4152..4400
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     4152..4172
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     4323..4342
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     enhancer        4453..4738
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 gggcgagggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc
       61 gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag
      121 cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc
      181 ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga
      241 cggcccttct cctccgggct gtaattagct gagcaagagg taagggttta agggatggtt
      301 ggttggtggg gtattaatgt ttaattacct ggagcacctg cctgaaatca ctttttttca
      361 ggttggaccg gtgccaccat ggtgagcaag ggcgaggagg ataacatggc catcatcaag
      421 gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc
      481 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc
      541 aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc
      601 aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag
      661 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag
      721 gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc
      781 ccctccgacg gccccgtaat gcagaaaaag accatgggct gggaggcctc ctccgagcgg
      841 atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac
      901 ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg
      961 cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc
     1021 atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg
     1081 tacaagtagg atccaccgga tctagataac tgatcataat cagccatacc acatttgtag
     1141 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     1201 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     1261 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     1321 aactcatcaa tgtatcttaa cgcggtgaac catcacccta atcaagtttt ttggggtcga
     1381 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     1441 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     1501 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     1561 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     1621 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     1681 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg
     1741 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     1801 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     1861 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     1921 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     1981 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     2041 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca
     2101 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     2161 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     2221 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     2281 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     2341 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     2401 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     2461 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     2521 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     2581 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg
     2641 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     2701 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     2761 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     2821 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     2881 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     2941 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     3001 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     3061 ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     3121 tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg
     3181 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     3241 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     3301 gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact
     3361 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     3421 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     3481 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3541 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3601 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     3661 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3721 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3781 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3841 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3901 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3961 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4021 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4081 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     4141 tgctcacatg tgagggccta tttcccatga ttccttcata tttgcatata cgatacaagg
     4201 ctgttagaga gataattgga attaatttga ctgtaaacac aaagatatta gtacaaaata
     4261 cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa
     4321 tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct
     4381 tgtggaaagg acgaaacacc gtaatttcta ctcttgtaga tggagacggg atcccgtctc
     4441 cttttttgta cccgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     4501 cccccgccca ttgacgtcaa tagtaacgcc aatagggact ttccattgac gtcaatgggt
     4561 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
     4621 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattgtgccc agtacatgac
     4681 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     4741 cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc cacccccaat
     4801 tttgtattta tttatttttt aattattttg tgcagcgatg ggggcggggg gggggg
//
