LOCUS       sequence-68413-e        6843 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination plasmid with mClover for Golden Gate reaction of TALE 
            assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6843)
  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 6843)
  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: pTALYM3
FEATURES             Location/Qualifiers
     source          1..6843
                     /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             join(2965..2967,2968..2970,2971..3681)
                     /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(3010..3031)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     3618..3639
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(3697..3715)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3698..3715)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             3917..3946
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3962..3979
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4002..4019)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4008..4232
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4242..4258)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4400..4855
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4487..4506)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4697..4718
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4881..4985
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4986..5054,5055..5846)
                     /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(5204..5223)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6017..6605
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6506..6525
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6759..6776
                     /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 agcaagggcg aggagctgtt caccggggtg
     3001 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtccgcggc
     3061 gagggcgagg gcgatgccac caacggcaag ctgaccctga agttcatctg caccaccggc
     3121 aagctgcccg tgccctggcc caccctcgtg accaccttcg gctacggcgt ggcctgcttc
     3181 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     3241 tacgtccagg agcgcaccat ctctttcaag gacgacggta cctacaagac ccgcgccgag
     3301 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     3361 gaggacggca acatcctggg gcacaagctg gagtacaact tcaacagcca caacgtctat
     3421 atcacggccg acaagcagaa gaacggcatc aaggctaact tcaagatccg ccacaacgtt
     3481 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     3541 cccgtgctgc tgcccgacaa ccactacctg agccatcagt ccaagctgag caaagacccc
     3601 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gattacacat
     3661 ggcatggacg agctgtacaa gtctagaggg ccctattcta tagtgtcacc taaatgctaa
     3721 gaattctgta catatgggca ccaaagaacc tgtaaacgtt atctttttaa attgaatgtg
     3781 cacaaataaa agtttggaaa agaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     3841 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa atacgcatca tacgctgtgg cgtacctcga
     3901 gtctagaggg cccttcgaac aaaaactcat ctcagaagag gatctgaata tgcataccgg
     3961 tcatcatcac catcaccatt gagtttaaac ccgctgatca gcctcgactg tgccttctag
     4021 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     4081 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     4141 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     4201 caggcatgct ggggatgcgg tgggctctat gggcggccgc cactggccgt cgttttacaa
     4261 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     4321 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     4381 agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4441 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     4501 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     4561 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     4621 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4681 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     4741 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     4801 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
     4861 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4921 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4981 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5041 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5101 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5161 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5221 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5281 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5341 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5401 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5461 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5521 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5581 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5641 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5701 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5761 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5821 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     5881 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5941 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6001 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6061 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6121 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     6181 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6241 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6301 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6361 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6421 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6481 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6541 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6601 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6661 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     6721 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     6781 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     6841 gct
//
