LOCUS       sequence-68419-e        7889 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TALE-mRuby2 against Mus spretus minor satellite sequence containing 
            two SNPs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7889)
  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 7889)
  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-01
FEATURES             Location/Qualifiers
     source          1..7889
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    50..71
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(86..91,92..109,110..116)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    124..140
                     /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     129..151
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     148..164
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     148..164
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        184..563
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        565..842
                     /label=chicken beta-actin promoter
     intron          843..1851
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1775..1797
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1864..1883
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1864..1882
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1902..1922
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4053..4763
                     /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             4963..4992
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5008..5025
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5048..5065)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5054..5278
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5288..5305)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5288..5304)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5297..5319)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5446..5901
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5533..5552)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5743..5764
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5927..6031
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6032..6100,6101..6892)
                     /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(6250..6269)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7063..7651
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7552..7571
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7805..7822
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
       61 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
      121 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgag gcgcgccgga
      181 ttcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
      241 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      301 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      361 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      421 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      481 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      541 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      601 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      661 cgatgggggc gggggggggg ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc
      721 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      781 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      841 cgggagtcgc tgcgttgcct tcgccccgtg ccccgctccg cgccgcctcg cgccgcccgc
      901 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
      961 cgggctgtaa ttagcgcttg gtttaatgac ggctcgtttc ttttctgtgg ctgcgtgaaa
     1021 gccttaaagg gctccgggag ggccctttgt gcggggggga gcggctcggg gggtgcgtgc
     1081 gtgtgtgtgt gcgtggggag cgccgcgtgc ggcccgcgct gcccggcggc tgtgagcgct
     1141 gcgggcgcgg cgcggggctt tgtgcgctcc gcgtgtgcgc gaggggagcg cggccggggg
     1201 cggtgccccg cggtgcgggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc
     1261 gtgggggggt gagcaggggg tgtgggcgcg gcggtcgggc tgtaaccccc ccctgcaccc
     1321 ccctccccga gttgctgagc acggcccggc ttcgggtgcg gggctccgtg cggggcgtgg
     1381 cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg
     1441 gccgcctcgg gccggggagg gctcggggga ggggcgcggc ggccccggag cgccggcggc
     1501 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
     1561 ggacttcctt tgtcccaaat ctggcggagc cgaaatctgg gaggcgccgc cgcaccccct
     1621 ctagcgggcg cgggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct
     1681 tcgtgcgtcg ccgcgccgcc gtccccttct ccatctccag cctcggggct gccgcagggg
     1741 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
     1801 ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca gatccttaat
     1861 taataatacg actcactata ggggccgcca ccatgagatc tcctaagaaa aagaggaagg
     1921 tgcaggtgga tctacgcacg ctcggctaca gtcagcagca gcaagagaag atcaaaccga
     1981 aggtgcgttc gacagtggcg cagcaccacg aggcactggt gggccatggg tttacacacg
     2041 cgcacatcgt tgcgctcagc caacacccgg cagcgttagg gaccgtcgct gtcacgtatc
     2101 agcacataat cacggcgttg ccagaggcga cacacgaaga catcgttggc gtcggcaaac
     2161 agtggtccgg cgcacgcgcc ctggaggcct tgctcacgga tgcgggggag ttgagaggtc
     2221 cgccgttaca gttggacaca ggccaacttg tgaagattgc aaaacgtggc ggcgtgaccg
     2281 caatggaggc agtgcatgca tcgcgcaatg cactgacggg tgcccccctg aacctgaccc
     2341 cggaccaagt ggtggctatc gccagcaacg gtggcggcaa gcaagcgctc gaaacggtgc
     2401 agcggctgtt gccggtgctg tgccaggacc atggcctgac tccggaccaa gtggtggcta
     2461 tcgccagcca cgatggcggc aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc
     2521 tgtgccagga ccatggcctg accccggacc aagtggtggc tatcgccagc aacattggcg
     2581 gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag gaccatggcc
     2641 tgaccccgga ccaagtggtg gctatcgcca gcaacggtgg cggcaagcaa gcgctcgaaa
     2701 cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg cctgactccg gaccaagtgg
     2761 tggctatcgc cagccacgat ggcggcaagc aagcgctcga aacggtgcag cggctgttgc
     2821 cggtgctgtg ccaggaccat ggcctgaccc cggaccaagt ggtggctatc gccagcaacg
     2881 gtggcggcaa gcaagcgctc gaaacggtgc agcggctgtt gccggtgctg tgccaggacc
     2941 atggcctgac cccggaccaa gtggtggcta tcgccagcaa cattggcggc aagcaagcgc
     3001 tcgaaacggt gcagcggctg ttgccggtgc tgtgccagga ccatggcctg accccggacc
     3061 aagtggtggc tatcgccagc aacattggcg gcaagcaagc gctcgaaacg gtgcagcggc
     3121 tgttgccggt gctgtgccag gaccatggcc tgactccgga ccaagtggtg gctatcgcca
     3181 gccacgatgg cggcaagcaa gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc
     3241 aggaccatgg cctgaccccg gaccaagtgg tggctatcgc cagcaacatt ggcggcaagc
     3301 aagcgctcga aacggtgcag cggctgttgc cggtgctgtg ccaggaccat ggcctgaccc
     3361 cggaccaagt ggtggctatc gccagcaacg gtggcggcaa gcaagcgctc gaaacggtgc
     3421 agcggctgtt gccggtgctg tgccaggacc atggcctgac cccggaccaa gtggtggcta
     3481 tcgccagcaa tcacggcggc aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc
     3541 tgtgccagga ccatggcctg accccggacc aagtggtggc tatcgccagc aacggtggcg
     3601 gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag gaccatggcc
     3661 tgaccccgga ccaagtggtg gctatcgcca gcaacggtgg cggcaagcaa gcgctcgaaa
     3721 cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg cctgaccccg gaccaagtgg
     3781 tggctatcgc cagccacgat ggcggcaagc aagcgctcga aagcattgtg gcccagctga
     3841 gccggcctga tccggcgttg gccgcgttga ccaacgacca cctcgtcgcc ttggcctgcc
     3901 tcggcggacg tcctgccatg gatgcagtga aaaagggatt gccgcacgcg ccggaattga
     3961 tcagaagagt caatcgccgt attggcgaac gcacgtccca tcgcgttgcg ggatccactg
     4021 gaggcggagg tggctccggc accgcctcct ccatggtgtc taagggcgaa gagctgatca
     4081 aggaaaatat gcgtatgaag gtggtcatgg aaggttcggt caacggccac caattcaaat
     4141 gcacaggtga aggagaaggc aatccgtaca tgggaactca aaccatgagg atcaaagtca
     4201 tcgagggagg acccctgcca tttgcctttg acattcttgc cacgtcgttc atgtatggca
     4261 gccgtacttt tatcaagtac ccgaaaggca ttcctgattt ctttaaacag tcctttcctg
     4321 agggttttac ttgggaaaga gttacgagat acgaagatgg tggagtcgtc accgtcatgc
     4381 aggacaccag ccttgaggat ggctgtctcg tttaccacgt ccaagtcaga ggggtaaact
     4441 ttccctccaa tggtcccgtg atgcagaaga agaccaaggg ttgggagcct aatacagaga
     4501 tgatgtatcc agcagatggt ggtctgaggg gatacactca tatggcactg aaagttgatg
     4561 gtggtggcca tctgtcttgc tctttcgtaa caacttacag gtcaaaaaag accgtcggga
     4621 acatcaagat gcccggtatc catgccgttg atcaccgcct ggaaaggtta gaggaaagtg
     4681 acaatgaaat gttcgtagta caacgcgaac acgcagttgc caagttcgcc gggcttggtg
     4741 gtgggatgga cgagctgtac aagtaagaat tctgtacata tgggcaccaa agaacctgta
     4801 aacgttatct ttttaaattg aatgtgcaca aataaaagtt tggaaaagaa aaaaaaaaaa
     4861 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaatac
     4921 gcatcatacg ctgtggcgta cctcgagtct agagggccct tcgaacaaaa actcatctca
     4981 gaagaggatc tgaatatgca taccggtcat catcaccatc accattgagt ttaaacccgc
     5041 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     5101 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     5161 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     5221 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatgggc
     5281 ggccgccact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     5341 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     5401 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg
     5461 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     5521 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     5581 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     5641 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     5701 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     5761 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     5821 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     5881 aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat
     5941 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     6001 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     6061 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     6121 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     6181 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     6241 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     6301 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     6361 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     6421 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     6481 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     6541 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     6601 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     6661 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     6721 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     6781 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     6841 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     6901 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6961 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     7021 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     7081 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     7141 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     7201 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     7261 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     7321 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     7381 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     7441 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     7501 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     7561 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     7621 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     7681 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     7741 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     7801 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     7861 cgcgcgttgg ccgattcatt aatgcagct
//
