LOCUS       sequence-207250-        4860 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses Mb crRNA and mCherry in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4860)
  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 4860)
  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: pTE3179
FEATURES             Location/Qualifiers
     source          1..4860
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          138..365
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      377..386
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             383..1093
                     /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(528..546)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    1222..1343
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1259..1278)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1313..1332
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1473..1492
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1606..1710
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1712..2069
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1712..1903
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1735..1755)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1920..2055
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2104..2898
                     /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             2104..2898
                     /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(2158..2177)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2768..2787
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3086..3105)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3130..3177
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3506..4094
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3995..4014
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4156..4404
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     4156..4176
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     4327..4346
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     enhancer        4458..4743
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 ggcggggcga ggggcggggc ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg
       61 gcgcgctccg aaagtttcct tttatggcga ggcggcggcg gcggcggccc tataaaaagc
      121 gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg
      181 ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc
      241 gggacggccc ttctcctccg ggctgtaatt agctgagcaa gaggtaaggg tttaagggat
      301 ggttggttgg tggggtatta atgtttaatt acctggagca cctgcctgaa atcacttttt
      361 ttcaggttgg accggtgcca ccatggtgag caagggcgag gaggataaca tggccatcat
      421 caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga
      481 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
      541 gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg
      601 ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc
      661 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac
      721 ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa
      781 cttcccctcc gacggccccg taatgcagaa aaagaccatg ggctgggagg cctcctccga
      841 gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa
      901 ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca
      961 gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta
     1021 caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga
     1081 gctgtacaag taggatccac cggatctaga taactgatca taatcagcca taccacattt
     1141 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     1201 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     1261 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     1321 tccaaactca tcaatgtatc ttaacgcggt gaaccatcac cctaatcaag ttttttgggg
     1381 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     1441 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     1501 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     1561 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     1621 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     1681 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     1741 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     1801 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     1861 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     1921 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     1981 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     2041 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     2101 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     2161 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     2221 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     2281 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     2341 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     2401 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     2461 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     2521 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     2581 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     2641 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     2701 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     2761 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     2821 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     2881 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     2941 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     3001 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     3061 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     3121 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     3181 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     3241 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     3301 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     3361 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     3421 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     3481 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     3541 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     3601 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     3661 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3721 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3781 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3841 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3901 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3961 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4021 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4081 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4141 cttttgctca catgtgaggg cctatttccc atgattcctt catatttgca tatacgatac
     4201 aaggctgtta gagagataat tggaattaat ttgactgtaa acacaaagat attagtacaa
     4261 aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt
     4321 aaaatggact atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat
     4381 atcttgtgga aaggacgaaa caccgaaatt tctactgttt gtagatggag acgttcgaac
     4441 gtctcctttt ttgtacccgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     4501 aacgaccccc gcccattgac gtcaatagta acgccaatag ggactttcca ttgacgtcaa
     4561 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     4621 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcattg tgcccagtac
     4681 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     4741 atggtcgagg tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc
     4801 ccaattttgt atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggggg
//
