LOCUS       sequence-207216-        4875 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses Lb crRNA and mCherry in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4875)
  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 4875)
  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: pTE4561
FEATURES             Location/Qualifiers
     source          1..4875
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          153..380
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      392..401
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             398..1108
                     /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(543..561)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    1237..1358
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1274..1293)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1328..1347
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1488..1507
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1621..1725
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1727..2084
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1727..1918
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1750..1770)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1935..2070
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2119..2913
                     /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             2119..2913
                     /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(2173..2192)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2783..2802
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3101..3120)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3145..3192
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3521..4109
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4010..4029
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4171..4419
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     4171..4191
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     4342..4361
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     enhancer        4473..4758
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 cgccaggcgg ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg
       61 cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc
      121 ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg
      181 tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc
      241 cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagctg agcaagaggt
      301 aagggtttaa gggatggttg gttggtgggg tattaatgtt taattacctg gagcacctgc
      361 ctgaaatcac tttttttcag gttggaccgg tgccaccatg gtgagcaagg gcgaggagga
      421 taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa
      481 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
      541 cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc
      601 tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt
      661 gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
      721 cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa
      781 gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaaaaaga ccatgggctg
      841 ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca
      901 gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc
      961 caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc
     1021 ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac
     1081 cggcggcatg gacgagctgt acaagtagga tccaccggat ctagataact gatcataatc
     1141 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     1201 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     1261 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     1321 tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcggtgaacc atcaccctaa
     1381 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     1441 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     1501 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     1561 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     1621 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     1681 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
     1741 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     1801 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     1861 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     1921 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     1981 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     2041 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     2101 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     2161 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     2221 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     2281 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     2341 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     2401 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     2461 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     2521 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     2581 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     2641 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     2701 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     2761 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     2821 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     2881 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     2941 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     3001 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     3061 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     3121 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     3181 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     3241 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     3301 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     3361 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     3421 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     3481 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     3541 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     3601 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     3661 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3721 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3781 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3841 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     3901 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     3961 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     4021 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     4081 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     4141 tggccttttg ctggcctttt gctcacatgt gagggcctat ttcccatgat tccttcatat
     4201 ttgcatatac gatacaaggc tgttagagag ataattggaa ttaatttgac tgtaaacaca
     4261 aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt
     4321 ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat
     4381 ttcttggctt tatatatctt gtggaaagga cgaaacaccg taatttctac taagtgtaga
     4441 tggagacggg atcccgtctc cttttttgta cccgttacat aacttacggt aaatggcccg
     4501 cctggctgac cgcccaacga cccccgccca ttgacgtcaa tagtaacgcc aatagggact
     4561 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     4621 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     4681 cattgtgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     4741 gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc
     4801 cccccctccc cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg
     4861 ggggcggggg ggggg
//
