LOCUS       sequence-215904-        5708 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Homology arms and linker-mEGFP sequence for C-terminus tagging of 
            human TNNI1, via excisable Cas9-excisable hPGK-mCherry selection 
            cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5708)
  AUTHORS   Roberts B, Hendershott MC, Arakaki J, Gerbin KA, Malik H, Nelson A, 
            Gehring J, Hookway C, Ludmann SA, Yang R, Haupt A, Grancharova T, 
            Valencia V, Fuqua MA, Tucker A, Rafelski SM, Gunawardane RN
  TITLE     Fluorescent Gene Tagging of Transcriptionally Silent Genes in 
            hiPSCs.
  JOURNAL   Stem Cell Reports. 2019 May 14;12(5):1145-1158. doi: 
            10.1016/j.stemcr.2019.03.001. Epub 2019 Apr 4.
  PUBMED    30956114
REFERENCE   2  (bases 1 to 5708)
  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"; doi: 
            "10.1016/j.stemcr.2019.03.001"; journalName: "Stem Cell Reports"; 
            date: "2019-05-14- 14"; volume: "12"; issue: "5"; pages: "1145-1158"
COMMENT     Sequence Label: AICSDP-34: TNNI1-mEGFP
FEATURES             Location/Qualifiers
     source          1..5708
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     polyA_signal    1088..1209
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1125..1144)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1179..1198
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     enhancer        1224..1603
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1605..1882
                     /label=chicken beta-actin promoter
     intron          1884..2901
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2825..2847
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2909..2928
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(3129..3147)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3397..3416
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3702..3926
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             join(3995..3997,3998..4711)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(4037..4058)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4298..4317)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4645..4666
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 caccctctgc cctaccagct ttccccttgt caacccctca catggcctca ggctggccca
       61 ggagctcctg ttggagacca gtcagcttga cagacagaac attctagaat gctttgggag
      121 accagtgtgg gaagaaagaa agaagatttg ctcccttcct tcaggctgtt cccagtctga
      181 cgaagaaact aggacataaa gaaagctaga ccccagacgg agaaatgaag taaaaatctt
      241 gttcccagtg tgaagataat gtggagattt agggtaatgg tggagatgac tgcctggtca
      301 ccacagggaa aggagacagc agctcatctt ggttatagcc acacacgagc atagacatta
      361 gagaactgag ttcaaccacc cagcaacaaa ttattcaaag ttggttgaat tcacagtata
      421 tgtgccccaa attgataggc tctgtaagga acagggatga aggaacagtc tctggcccaa
      481 gaagcttcag aggagaagag ataagccaga caatatactg aggaaatgga aaagcagccg
      541 catttgcagg ggaagacttc ttggaagagg tggccttgag cctgctgagt gccagaccag
      601 gagggtggca tggagccttg gaggaggttt tgctgaggtg cactgaggtg ggaactagcc
      661 ccatagctgc cactttcacc cagtgctccc aagccacaca gcttcctttt tcctgtgggg
      721 cctcggcttg gcctgtgtcc acctgcctgg ctgagtccta ggaccttacc agctggggag
      781 gggcatcagg atttcttatc atccgaggag actggtttcc tatcacccag cttctcttga
      841 gctggggggc cgtccagcct ggtaagtccc tgatgtgtgt gccacttgtc tacaggagcg
      901 gcctgtggag gtgggtgact ggaggaagaa cgtggaggcc atgtctggca tggaaggccg
      961 gaagaagatg tttgatgccg ccaagtctcc gacctcacaa tccgggtccg ggtcccccgg
     1021 agaggttccc gacataccaa taacttcgta tagtgtagga tatacgaagt tatattagta
     1081 ttccgataac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     1141 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     1201 tgtatcttat catgtctggt ctcgacattg attattgact agttattaat agtaatcaat
     1261 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     1321 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     1381 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     1441 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     1501 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     1561 tacttggcag tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac
     1621 gttctgcttc actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat
     1681 tttttaatta ttttgtgcag cgatgggggc gggggggggg ggggggcgcg cgccaggcgg
     1741 ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg cagccaatca
     1801 gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa
     1861 aaagcgaagc gcgcggcggg cggggagtcg ctgcgacgct gccttcgccc cgtgccccgc
     1921 tccgccgccg cctcgcgccg cccgccccgg ctctgactga ccgcgttact cccacaggtg
     1981 agcgggcggg acggcccttc tcctccgggc tgtaattagc gcttggttta atgacggctt
     2041 gtttcttttc tgtggctgcg tgaaagcctt gaggggctcc gggagggccc tttgtgcggg
     2101 gggagcggct cggggggtgc gtgcgtgtgt gtgtgcgtgg ggagcgccgc gtgcggctcc
     2161 gcgctgcccg gcggctgtga gcgctgcggg cgcggcgcgg ggctttgtgc gctccgcagt
     2221 gtgcgcgagg ggagcgcggc cgggggcggt gccccgcggt gcgggggggg ctgcgagggg
     2281 aacaaaggct gcgtgcgggg tgtgtgcgtg ggggggtgag cagggggtgt gggcgcgtcg
     2341 gtcgggctgc aaccccccct gcacccccct ccccgagttg ctgagcacgg cccggcttcg
     2401 ggtgcggggc tccgtacggg gcgtggcgcg gggctcgccg tgccgggcgg ggggtggcgg
     2461 caggtggggg tgccgggcgg ggcggggccg cctcgggccg gggagggctc gggggagggg
     2521 cgcggcggcc cccggagcgc cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt
     2581 ttatggtaat cgtgcgagag ggcgcaggga cttcctttgt cccaaatctg tgcggagccg
     2641 aaatctggga ggcgccgccg caccccctct agcgggcgcg gggcgaagcg gtgcggcgcc
     2701 ggcaggaagg aaatgggcgg ggagggcctt cgtgcgtcgc cgcgccgccg tccccttctc
     2761 cctctccagc ctcggggctg tccgcggggg gacggctgcc ttcggggggg acggggcagg
     2821 gcggggttcg gcttctggcg tgtgaccggc ggctctagag cctctgctaa ccatgttcat
     2881 gccttcttct ttttcctaca gctcctgggc aacgtgctgg ttattgtgct gtctcatcat
     2941 tttggcaaag aattccgcca ccggggatcc accggagctt accatggtct ccaaaggaga
     3001 agaagataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc
     3061 cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
     3121 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct
     3181 gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga
     3241 ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
     3301 cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa
     3361 ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat
     3421 gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat
     3481 caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta
     3541 caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat
     3601 cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca
     3661 ctccaccggc ggaatggatg aactctataa atgaggtact cctgtgcctt ctagttgcca
     3721 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     3781 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     3841 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     3901 tgctggggat gcggtgggct ctatggaaat aacttcgtat agtgtaggat atacgaagtt
     3961 atataggtat gtcgggaacc tctccgggtc cggggtgagc aagggcgagg agctgttcac
     4021 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     4081 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     4141 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     4201 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     4261 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     4321 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     4381 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     4441 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     4501 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     4561 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtcca agctgagcaa
     4621 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     4681 cactctcggc atggacgagc tgtacaagta gaggtaccta gagaccagct cccttctctg
     4741 gggtccatca gagtctagag gataatgagg gagtccacaa tgggcaggcg ggttgtactt
     4801 agtgtggtgc acggatgcac ggactaaggc accgatgaga ctgatttgag tcggctctcc
     4861 tgggctcagt cctggctcca gcaccatgat acttgggcat gtcatttctg tttggcggcc
     4921 ctttttgttc ccattatcta tgaaattggg ggctgtatca aatcagtggt tctccaagtg
     4981 tggtcctggg accagcagcg tctacatcac ctgggatgca aatgatcagc ccccacccca
     5041 gatccactaa attggaaact ctggggtgag gccagccttc taggtgattc tgatgcacat
     5101 gagagtttga gaaccactag gttagagggt ccctgggatc ccttcatctc tgactttctg
     5161 agaatctatg cacttttaaa tacctcctgg tgatgtcctt tatttgtgct ccctgcgaag
     5221 tacccagcac tcacagtacc tgttcccaag gagctaccac tcacagaagc taagacttcc
     5281 caccatctga ggaaatagct ggagaaactg ggcaaagcaa ggaaatcagg ccacgtaaaa
     5341 ccctctacgg aagtgctgag agaatgtgca atagggcagg catccaccag acagcactgg
     5401 tcagagcagg ctttctggag aggaggagct ctctgaggat gcgacgagcg agtgttagct
     5461 gataaaggga agggcatgtt gaagggctga tggtgcaaca gaagcatgga ggtgggaatt
     5521 aagcagccac agagtagcat ggcaaggcta ggtaagagct gagggccgca gtgtggagga
     5581 gcggtgtgag gggtcccaaa aggggagcaa ctgtgcccga gggccgagaa gctgaggttt
     5641 tgtaggagcc cttatgaatg aagatccagt ccggggtgcg ctagggctga gaagccctgg
     5701 ggactgca
//
