LOCUS       sequence-68420-e        7890 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TALE-mRuby2 against Mus spretus/musculus minor satellite sequence .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7890)
  AUTHORS   Miyanari Y, Ziegler-Birling C, Torres-Padilla ME
  TITLE     Live visualization of chromatin dynamics with fluorescent TALEs.
  JOURNAL   Nat Struct Mol Biol. 2013 Oct 6. doi: 10.1038/nsmb.2680.
  PUBMED    24096363
REFERENCE   2  (bases 1 to 7890)
  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: "Nat Struct 
            Mol Biol. 2013 Oct 6. doi: 10.1038/nsmb.2680."
COMMENT     Sequence Label: pTALYM4SpMi-05
FEATURES             Location/Qualifiers
     source          1..7890
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1870..2580
                     /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"
     CDS             2780..2809
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2825..2842
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2865..2882)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2871..3095
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3105..3122)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3105..3121)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3114..3136)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3263..3718
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3350..3369)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3560..3581
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3744..3848
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3849..3917,3918..4709)
                     /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     complement(4067..4086)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4880..5468
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5369..5388
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5622..5639
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5756..5777
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5792..5797,5798..5815,5816..5822)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5830..5846
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5835..5857
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5854..5870
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5854..5870
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        5890..6269
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6271..6548
                     /label=chicken beta-actin promoter
     intron          6549..7558
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     7482..7504
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     7571..7590
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7571..7589
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7609..7629
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 gaggccttgc tcacggatgc gggggagttg agaggtccgc cgttacagtt ggacacaggc
       61 caacttgtga agattgcaaa acgtggcggc gtgaccgcaa tggaggcagt gcatgcatcg
      121 cgcaatgcac tgacgggtgc ccccctgaac ctgaccccgg accaagtggt ggctatcgcc
      181 agccacgatg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc
      241 caggaccatg gcctgacccc ggaccaagtg gtggctatcg ccagcaacat tggcggcaag
      301 caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgacc
      361 ccggaccaag tggtggctat cgccagcaac ggtggcggca agcaagcgct cgaaacggtg
      421 cagcggctgt tgccggtgct gtgccaggac catggcctga ccccggacca agtggtggct
      481 atcgccagca acggtggcgg caagcaagcg ctcgaaacgg tgcaacggct gttgccggtg
      541 ctgtgccagg accatggcct gaccccggac caagtggtgg ctatcgccag caaccacggc
      601 ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc
      661 ctgaccccgg accaagtggt ggctatcgcc agcaacggtg gcggcaagca agcgctcgaa
      721 acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgacccc ggaccaagtg
      781 gtggctatcg ccagcaacat tggcggcaag caagcgctcg aaacggtgca gcggctgttg
      841 ccggtgctgt gccaggacca tggcctgacc ccggaccaag tggtggctat cgccagcaac
      901 attggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct gtgccaggac
      961 catggcctga ctccggacca agtggtggct atcgccagcc acgatggcgg caagcaagcg
     1021 ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct gaccccggac
     1081 caagtggtgg ctatcgccag caacggtggc ggcaagcaag cgctcgaaac ggtgcagcgg
     1141 ctgttgccgg tgctgtgcca ggaccatggc ctgaccccgg accaagtggt ggctatcgcc
     1201 agccacgatg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc
     1261 caggaccatg gcctgacccc ggaccaagtg gtggctatcg ccagcaacat tggcggcaag
     1321 caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca tggcctgacc
     1381 ccggaccaag tggtggctat cgccagcaac ggtggcggca agcaagcgct cgaaacggtg
     1441 cagcggctgt tgccggtgct gtgccaggac catggcctga ccccggacca agtggtggct
     1501 atcgccagca acggtggcgg caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg
     1561 ctgtgccagg accatggcct gaccccggac caagtggtgg ctatcgccag caaccacggc
     1621 ggcaagcaag cgctcgaaag cattgtggcc cagctgagcc ggcctgatcc ggcgttggcc
     1681 gcgttgacca acgaccacct cgtcgccttg gcctgcctcg gcggacgtcc tgccatggat
     1741 gcagtgaaaa agggattgcc gcacgcgccg gaattgatca gaagagtcaa tcgccgtatt
     1801 ggcgaacgca cgtcccatcg cgttgcggga tccactggag gcggaggtgg ctccggcacc
     1861 gcctcctcca tggtgtctaa gggcgaagag ctgatcaagg aaaatatgcg tatgaaggtg
     1921 gtcatggaag gttcggtcaa cggccaccaa ttcaaatgca caggtgaagg agaaggcaat
     1981 ccgtacatgg gaactcaaac catgaggatc aaagtcatcg agggaggacc cctgccattt
     2041 gcctttgaca ttcttgccac gtcgttcatg tatggcagcc gtacttttat caagtacccg
     2101 aaaggcattc ctgatttctt taaacagtcc tttcctgagg gttttacttg ggaaagagtt
     2161 acgagatacg aagatggtgg agtcgtcacc gtcatgcagg acaccagcct tgaggatggc
     2221 tgtctcgttt accacgtcca agtcagaggg gtaaactttc cctccaatgg tcccgtgatg
     2281 cagaagaaga ccaagggttg ggagcctaat acagagatga tgtatccagc agatggtggt
     2341 ctgaggggat acactcatat ggcactgaaa gttgatggtg gtggccatct gtcttgctct
     2401 ttcgtaacaa cttacaggtc aaaaaagacc gtcgggaaca tcaagatgcc cggtatccat
     2461 gccgttgatc accgcctgga aaggttagag gaaagtgaca atgaaatgtt cgtagtacaa
     2521 cgcgaacacg cagttgccaa gttcgccggg cttggtggtg ggatggacga gctgtacaag
     2581 taagaattct gtacatatgg gcaccaaaga acctgtaaac gttatctttt taaattgaat
     2641 gtgcacaaat aaaagtttgg aaaagaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     2701 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaatacgca tcatacgctg tggcgtacct
     2761 cgagtctaga gggcccttcg aacaaaaact catctcagaa gaggatctga atatgcatac
     2821 cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga ctgtgccttc
     2881 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     2941 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     3001 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     3061 tagcaggcat gctggggatg cggtgggctc tatgggcggc cgccactggc cgtcgtttta
     3121 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     3181 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     3241 cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3301 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3361 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3421 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3481 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3541 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3601 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     3661 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
     3721 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3781 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3841 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     3901 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     3961 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     4021 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     4081 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     4141 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     4201 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     4261 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     4321 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     4381 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     4441 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     4501 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     4561 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     4621 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     4681 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     4741 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     4801 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     4861 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4921 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4981 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     5041 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5101 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5161 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5221 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5281 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5341 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5401 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     5461 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     5521 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     5581 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     5641 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     5701 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     5761 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     5821 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgaggcgc
     5881 gccggattcg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
     5941 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     6001 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     6061 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     6121 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     6181 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     6241 tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc
     6301 tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt
     6361 gtgcagcgat gggggcgggg gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg
     6421 aggggcgggg cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc
     6481 gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc
     6541 ggcgggcggg agtcgctgcg ttgccttcgc cccgtgcccc gctccgcgcc gcctcgcgcc
     6601 gcccgccccg gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt
     6661 ctcctccggg ctgtaattag cgcttggttt aatgacggct cgtttctttt ctgtggctgc
     6721 gtgaaagcct taaagggctc cgggagggcc ctttgtgcgg gggggagcgg ctcggggggt
     6781 gcgtgcgtgt gtgtgtgcgt ggggagcgcc gcgtgcggcc cgcgctgccc ggcggctgtg
     6841 agcgctgcgg gcgcggcgcg gggctttgtg cgctccgcgt gtgcgcgagg ggagcgcggc
     6901 cgggggcggt gccccgcggt gcgggggggc tgcgagggga acaaaggctg cgtgcggggt
     6961 gtgtgcgtgg gggggtgagc agggggtgtg ggcgcggcgg tcgggctgta acccccccct
     7021 gcacccccct ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtgcggg
     7081 gcgtggcgcg gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg
     7141 ggcggggccg cctcgggccg gggagggctc gggggagggg cgcggcggcc ccggagcgcc
     7201 ggcggctgtc gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg
     7261 gcgcagggac ttcctttgtc ccaaatctgg cggagccgaa atctgggagg cgccgccgca
     7321 ccccctctag cgggcgcggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     7381 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     7441 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     7501 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagat
     7561 ccttaattaa taatacgact cactataggg gccgccacca tgagatctcc taagaaaaag
     7621 aggaaggtgc aggtggatct acgcacgctc ggctacagtc agcagcagca agagaagatc
     7681 aaaccgaagg tgcgttcgac agtggcgcag caccacgagg cactggtggg ccatgggttt
     7741 acacacgcgc acatcgttgc gctcagccaa cacccggcag cgttagggac cgtcgctgtc
     7801 acgtatcagc acataatcac ggcgttgcca gaggcgacac acgaagacat cgttggcgtc
     7861 ggcaaacagt ggtccggcgc acgcgccctg
//
