LOCUS       sequence-68417-e        7931 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TALE-mClover against mouse major satellite sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7931)
  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 7931)
  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: pTALYM3B15
FEATURES             Location/Qualifiers
     source          1..7931
                     /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             join(4053..4055,4056..4058,4059..4769)
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from mClover2, with improved photostability (Bajar et al., 
                     2003)"
                     /label=mClover3
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(4098..4119)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     4706..4727
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(4785..4803)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4786..4803)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             5005..5034
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5050..5067
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5090..5107)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5096..5320
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5330..5347)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5330..5346)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5339..5361)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5488..5943
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5575..5594)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5785..5806
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5969..6073
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6074..6142,6143..6934)
                     /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(6292..6311)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7105..7693
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7594..7613
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7847..7864
                     /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 cccggaccaa gtggtggcta
     2461 tcgccagcaa caagggcggc aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc
     2521 tgtgccagga ccatggcctg actccggacc aagtggtggc tatcgccagc cacgatggcg
     2581 gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag gaccatggcc
     2641 tgactccgga ccaagtggtg gctatcgcca gccacgatgg cggcaagcaa gcgctcgaaa
     2701 cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg cctgaccccg gaccaagtgg
     2761 tggctatcgc cagcaacatt 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 aacggtggcg gcaagcaagc gctcgaaacg gtgcagcggc
     3121 tgttgccggt gctgtgccag gaccatggcc tgaccccgga ccaagtggtg gctatcgcca
     3181 gcaacggtgg cggcaagcaa gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc
     3241 aggaccatgg cctgactccg gaccaagtgg tggctatcgc cagccacgat ggcggcaagc
     3301 aagcgctcga aacggtgcag cggctgttgc cggtgctgtg ccaggaccat ggcctgaccc
     3361 cggaccaagt ggtggctatc gccagccacg atggcggcaa gcaagcgctc gaaacggtgc
     3421 agcggctgtt gccggtgctg tgccaggacc atggcctgac cccggaccaa gtggtggcta
     3481 tcgccagcaa cattggcggc aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc
     3541 tgtgccagga ccatggcctg actccggacc aagtggtggc tatcgccagc cacgatggcg
     3601 gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag gaccatggcc
     3661 tgaccccgga ccaagtggtg gctatcgcca gcaacaaggg cggcaagcaa gcgctcgaaa
     3721 cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg cctgaccccg gaccaagtgg
     3781 tggctatcgc cagcaacggt 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 ccatggtgag caagggcgag gagctgttca
     4081 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     4141 tccgcggcga gggcgagggc gatgccacca acggcaagct gaccctgaag ttcatctgca
     4201 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccttcggc tacggcgtgg
     4261 cctgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     4321 ccgaaggcta cgtccaggag cgcaccatct ctttcaagga cgacggtacc tacaagaccc
     4381 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     4441 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaacttc aacagccaca
     4501 acgtctatat cacggccgac aagcagaaga acggcatcaa ggctaacttc aagatccgcc
     4561 acaacgttga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     4621 gcgacggccc cgtgctgctg cccgacaacc actacctgag ccatcagtcc aagctgagca
     4681 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     4741 ttacacatgg catggacgag ctgtacaagt ctagagggcc ctattctata gtgtcaccta
     4801 aatgctaaga attctgtaca tatgggcacc aaagaacctg taaacgttat ctttttaaat
     4861 tgaatgtgca caaataaaag tttggaaaag aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     4921 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaat acgcatcata cgctgtggcg
     4981 tacctcgagt ctagagggcc cttcgaacaa aaactcatct cagaagagga tctgaatatg
     5041 cataccggtc atcatcacca tcaccattga gtttaaaccc gctgatcagc ctcgactgtg
     5101 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     5161 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     5221 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     5281 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg gcggccgcca ctggccgtcg
     5341 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     5401 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     5461 agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
     5521 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     5581 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     5641 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     5701 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     5761 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     5821 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     5881 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     5941 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     6001 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     6061 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     6121 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     6181 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     6241 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     6301 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     6361 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     6421 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     6481 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     6541 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     6601 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     6661 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     6721 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     6781 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     6841 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     6901 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     6961 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     7021 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     7081 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     7141 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     7201 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     7261 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     7321 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     7381 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     7441 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     7501 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     7561 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     7621 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     7681 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     7741 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     7801 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     7861 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     7921 ttaatgcagc t
//
