LOCUS       sequence-68414-e        6801 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination plasmid with mRuby2 for Golden Gate reaction of TALE 
            assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6801)
  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 6801)
  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: pTALYM4
FEATURES             Location/Qualifiers
     source          1..6801
                     /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     148..164
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     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"
     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             complement(2226..2633)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLQGG
                     TRQCIARCMHCLHCGHAATFCNLHKLACVQL"
     primer_bind     2498..2514
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2524..2542
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2590..2610)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2590..2608)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2629..2645)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2653..2669
                     /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)."
     promoter        complement(join(2677..2683,2684..2701,2702..2707))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2722..2743
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             2965..3675
                     /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             3875..3904
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3920..3937
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3960..3977)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3966..4190
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4200..4216)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4358..4813
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4445..4464)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4655..4676
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4839..4943
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4944..5012,5013..5804)
                     /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(5162..5181)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5975..6563
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6464..6483
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6717..6734
                     /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 gagacgggcg
     2341 ccgctacagg gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg
     2401 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     2461 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc
     2521 gcgtaatacg actcactata gggcgaattg ggtaccgggc cccccctcga ggtcctccag
     2581 cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt
     2641 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     2701 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     2761 gcccgctttc caccggtcgt ctccaacgac cacctcgtcg ccttggcctg cctcggcgga
     2821 cgtcctgcca tggatgcagt gaaaaaggga ttgccgcacg cgccggaatt gatcagaaga
     2881 gtcaatcgcc gtattggcga acgcacgtcc catcgcgttg cgggatccac tggaggcgga
     2941 ggtggctccg gcaccgcctc ctccatggtg tctaagggcg aagagctgat caaggaaaat
     3001 atgcgtatga aggtggtcat ggaaggttcg gtcaacggcc accaattcaa atgcacaggt
     3061 gaaggagaag gcaatccgta catgggaact caaaccatga ggatcaaagt catcgaggga
     3121 ggacccctgc catttgcctt tgacattctt gccacgtcgt tcatgtatgg cagccgtact
     3181 tttatcaagt acccgaaagg cattcctgat ttctttaaac agtcctttcc tgagggtttt
     3241 acttgggaaa gagttacgag atacgaagat ggtggagtcg tcaccgtcat gcaggacacc
     3301 agccttgagg atggctgtct cgtttaccac gtccaagtca gaggggtaaa ctttccctcc
     3361 aatggtcccg tgatgcagaa gaagaccaag ggttgggagc ctaatacaga gatgatgtat
     3421 ccagcagatg gtggtctgag gggatacact catatggcac tgaaagttga tggtggtggc
     3481 catctgtctt gctctttcgt aacaacttac aggtcaaaaa agaccgtcgg gaacatcaag
     3541 atgcccggta tccatgccgt tgatcaccgc ctggaaaggt tagaggaaag tgacaatgaa
     3601 atgttcgtag tacaacgcga acacgcagtt gccaagttcg ccgggcttgg tggtgggatg
     3661 gacgagctgt acaagtaaga attctgtaca tatgggcacc aaagaacctg taaacgttat
     3721 ctttttaaat tgaatgtgca caaataaaag tttggaaaag aaaaaaaaaa aaaaaaaaaa
     3781 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaat acgcatcata
     3841 cgctgtggcg tacctcgagt ctagagggcc cttcgaacaa aaactcatct cagaagagga
     3901 tctgaatatg cataccggtc atcatcacca tcaccattga gtttaaaccc gctgatcagc
     3961 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     4021 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     4081 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     4141 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg gcggccgcca
     4201 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     4261 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     4321 ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
     4381 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4441 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4501 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4561 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4621 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4681 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     4741 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     4801 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     4861 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4921 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     4981 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     5041 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     5101 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     5161 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     5221 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     5281 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5341 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     5401 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5461 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     5521 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     5581 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     5641 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     5701 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     5761 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     5821 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5881 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5941 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6001 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6061 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6121 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6181 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6241 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6301 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6361 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6421 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6481 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6541 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6601 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6661 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     6721 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     6781 ggccgattca ttaatgcagc t
//
