LOCUS       sequence-362448-        5719 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Repair template for the N-terminal tagging of myosin heavy chain 10 
            with mRuby2 in human cells using CRISPR/Cas9..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5719)
  AUTHORS   Husser MC, Ozugergin I, Resta T, Martin VJJ, Piekny AJ
  TITLE     Cytokinetic diversity in mammalian cells is revealed by the 
            characterization of endogenous anillin, Ect2 and RhoA.
  JOURNAL   Open Biol. 2022 Nov;12(11):220247. doi: 10.1098/rsob.220247. Epub 
            2022 Nov 23.
  PUBMED    36416720
REFERENCE   2  (bases 1 to 5719)
  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: "Open 
            Biol."; date: "2022-11"; volume: "12(11):220247. doi"; pages: " 
            10.1098/rsob.220247. Epub 2022 Nov 23"
COMMENT     Sequence Label: pODN-mR2-MYH10
FEATURES             Location/Qualifiers
     source          1..5719
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(171..962,963..1031))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     794..813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1032..1136)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1204..1222
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     1528..1547
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1528..1546
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2592..3302
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     promoter        complement(join(4774..4780,4781..4798,4799..4804))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    4819..4840
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4960..4977)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5131..5719)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5211..5230)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1141 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     1201 aaaaataggc gtatcacgag gccgcccctg cagccgaatt atattatttt tgccaaataa
     1261 tttttaacaa aagctctgaa gtcttcttca tttaaattct tagatgatac ttcatctgga
     1321 aaattgtccc aattagtagc atcacgctgt gagtaagttc taaaccattt ttttattgtt
     1381 gtattatctc taatcttact actcgatgag ttttcggtat tatctctatt tttaacttgg
     1441 agcaggttcc attcattgtt tttttcatca tagtgaataa aatcaactgc tttaacactt
     1501 gtgcctgaac accatatcca tccggcgtaa tacgactcac tatagggaga gcggccgcca
     1561 gatcttccgg atggctcgag tttttcagca agataagtga tggagcttcc tctgctagcc
     1621 ctttgtgagc caatggtaaa tgggtgctaa ataaaacaac taggtcttga gatacattaa
     1681 ttgtaaatgt cacagaacca gtactttcct caatgtggct aagatagttg atggttcctt
     1741 tttcttctgc actggtcaga ccatatctgg gctatgatgt ttgcttctgg gccacacact
     1801 ttagagggaa gacaagcagc atgttggagt ctgtttaggc gagagatccg gcaggggaag
     1861 gagtcttggt gaatgagggg tggaggagct gcaggatggg aatagaggcc tgaactgcta
     1921 ccatgacata ttcaaaaggc tgccgtgtga agccaagtta tgcttgtctt ttgtggtccc
     1981 agttgatcac attaagacct catggggcca tagaaaagct agagggagac tgatttgggt
     2041 tattcataag aagaacttta agtctgttat ctgagggtag aatgagaggc atgttttaga
     2101 ttctttagat tctttactct tctgacaatc atgtgttttt gtagctgttt ccttgtggtc
     2161 atattaattc tggtaccact tcatgaacct tttattaccc atctttgttt tctttttttt
     2221 tttcccttct taactccctg tttaatttgt ggtgagggtg aaagaggaga taaagaaaaa
     2281 aaagggtcaa cttgtaactt tgccttttct tttcttttct tttctttttt ttgccctcag
     2341 taactgaggg caaacccatc agacaaccag agccataatt tgtggtcacc gctgaaattt
     2401 accttggaaa ctcggttagt atggctgtga agaggtatac cccagctcct taacacagag
     2461 ttaatgctta atctaaggtt ttaagtttct tagaaaagaa aaacgtgtac attcttttgt
     2521 ttcttaaaca tctaattttg ccctcctcct cttctcttag aggcaattgc ttttggatcg
     2581 ttccatttac aatggtgtct aagggcgaag agctgatcaa ggaaaatatg cgtatgaagg
     2641 tggtcatgga aggttcggtc aacggccacc aattcaaatg cacaggtgaa ggagaaggca
     2701 atccgtacat gggaactcaa accatgagga tcaaagtcat cgagggagga cccctgccat
     2761 ttgcctttga cattcttgcc acgtcgttca tgtatggcag ccgtactttt atcaagtacc
     2821 cgaaaggcat tcctgatttc tttaaacagt cctttcctga gggttttact tgggaaagag
     2881 ttacgagata cgaagatggt ggagtcgtca ccgtcatgca ggacaccagc cttgaggatg
     2941 gctgtctcgt ttaccacgtc caagtcagag gggtaaactt tccctccaat ggtcccgtga
     3001 tgcagaagaa aaccaagggt tgggagccta atacagagat gatgtatcca gcagatggtg
     3061 gtctgagggg atacactcat atggcactga aagttgatgg tggtggccat ctgtcttgct
     3121 ctttcgtaac aacttacagg tcaaaaaaga ccgtcgggaa catcaagatg cccggtatcc
     3181 atgccgttga tcaccgcctg gaaaggttag aggaaagtga caatgaaatg ttcgtagtac
     3241 aacgcgaaca cgcagttgcc aagttcgccg ggcttggtgg tgggatggac gagctgtaca
     3301 agtacagcga tctggagctg aagctgcgga tccctatggc acaaaggaca gggctggagg
     3361 atccagagag gtacctgttc gtcgatcgtg ctgtcatcta caaccctgcc actcaagctg
     3421 attggacagc taaaaagcta gtgtggattc catcagaacg ccatggtttt gaggcagcta
     3481 gtatcaaaga agaacgggga gatgaagtta tggtggagtt ggcagagaat ggaaagaaag
     3541 caatggtcaa caaagatgat attcagaaga tgaacccacc taagttttcc aaggtggagg
     3601 atatggcaga attgacatgc ttgaatgaag cttccgtttt acataatctg aaggatcgct
     3661 actattcagg actaatctat gtaagtattt cttccaaata atcatgtgaa gtggtagcta
     3721 ggaattaatg taaattatac atcttgtcat aatcaaatga gaatgtggaa tacccaaact
     3781 ctctgtttaa catttctatt tctctttaag atagaaagat ttgttgcttg cttacccatg
     3841 tcttgctttt ctttgaatct taacacatta agtttaaata atacaggctg caattacata
     3901 taataaaatg gcatttgaag acttttgtag tggtcttctg gagcataata aggtgggaga
     3961 gagcatgtaa caggaagacc agaaggttta ataaggtaaa gagagttgca ttaattggac
     4021 gcagacagca aaacggtcaa aaatcaagtg catacccaag agtaaagtgg aggggctgta
     4081 agctgagaaa tttctgtgga cagcatgaac agcttcactg gatgtagtag ggaagtagga
     4141 aagatgaatg ctgaggtttt taagaggaaa caattagggt aagattgagg ctggctgggg
     4201 tcgtcctgtg gttagcagct gacatgaatg ttggagtcac cgactttgtc actgaccatg
     4261 tagaagaagt tattgaaaat cataagggat aatgtagaga gggataatgt agagaggaaa
     4321 aatgtaagcc agatactaaa tctttctaga agatctccta caatattctc agctgccatg
     4381 gaaaatcgat gttcttcttt tattctctca agattttcag gctgtatatt aaaacttata
     4441 ttaagaacta tgctaaccac ctcatcagga accgttgtag gtggcgtggg ttttcttggc
     4501 aatcgactct catgaaaact acgagctaaa tattcaatat gttcctcttg accaacttta
     4561 ttctgcattt tttttgaacg aggtttagag caagcttcag gaaactgaga caggaatttt
     4621 attaaaaatt taaattttga agaaagttca gggttaatag catccatttt ttgctttgca
     4681 agttcctcag cattcttaac aaaagacgtc tcttttgaca tgtttaaagt ttaaacctcc
     4741 tgtgtgaaat tattatccgc tcataattcc acacattata cgagccggaa gcataaagtg
     4801 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     4861 aattgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4921 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4981 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5041 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5101 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5161 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5221 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5281 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5341 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5401 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5461 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5521 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5581 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5641 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5701 cagaaaaaaa ggatctcaa
//
