LOCUS       sequence-199271-        5871 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for IvT reaction with T3. Encodes LINE-1 specific TALE 
            sequence followed by VP64 activator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5871)
  AUTHORS   Jachowicz JW, Bing X, Pontabry J, Boskovic A, Rando OJ, 
            Torres-Padilla ME
  TITLE     LINE-1 activation after fertilization regulates global chromatin 
            accessibility in the early mouse embryo.
  JOURNAL   Nat Genet. 2017 Aug 28. doi: 10.1038/ng.3945.
  PUBMED    28846101
REFERENCE   2  (bases 1 to 5871)
  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 Genet. 
            2017 Aug 28. doi: 10.1038/ng.3945."
COMMENT     Sequence Label: TAL-L1-1.3-VP64
FEATURES             Location/Qualifiers
     source          1..5871
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..20
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     5'UTR           45..87
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     CDS             110..136
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             143..163
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2567..2590
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2594..2614
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2639..2788
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     3'UTR           2847..2972
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(3060..3079)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3061..3079)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3086..3103)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3086..3102)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3095..3117)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3244..3699)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3381..3402)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3593..3612
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3726..3830
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3831..3899,3900..4691)
                     /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(4049..4068)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4862..5450
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5351..5370
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5604..5621
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5738..5759
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5774..5779,5780..5797,5798..5804)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5812..5828
                     /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     5817..5839
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5836..5852
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5836..5852
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aaattaaccc tcactaaagg gaacaaaagc tggagctaag cttgcttgtt ctttttgcag
       61 aagctcagaa taaacgctca actttggcag atccgctacc gccgccatgt acccatacga
      121 tgttccagat tacgctagat ctcctaagaa aaagaggaag gtgcaggtgg atctacgcac
      181 gctcggctac agtcagcagc agcaagagaa gatcaaaccg aaggtgcgtt cgacagtggc
      241 gcagcaccac gaggcactgg tgggccatgg gtttacacac gcgcacatcg ttgcgctcag
      301 ccaacacccg gcagcgttag ggaccgtcgc tgtcacgtat cagcacataa tcacggcgtt
      361 gccagaggcg acacacgaag acatcgttgg cgtcggcaaa cagtggtccg gcgcacgcgc
      421 cctggaggcc ttgctcacgg atgcggggga gttgagaggt ccgccgttac agttggacac
      481 aggccaactt gtgaagattg caaaacgtgg cggcgtgacc gcaatggagg cagtgcatgc
      541 atcgcgcaat gcactgacgg gtgcccccct gaacctgacc ccggaccaag tggtggctat
      601 cgccagcaac cacggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct
      661 gtgccaggac catggcctga ctccggacca agtggtggct atcgccagcc acgatggcgg
      721 caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct
      781 gaccccggac caagtggtgg ctatcgccag caacggtggc ggcaagcaag cgctcgaaac
      841 ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc ctgactccgg accaagtggt
      901 ggctatcgcc agccacgatg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc
      961 ggtgctgtgc caggaccatg gcctgacccc ggaccaagtg gtggctatcg ccagcaacgg
     1021 tggcggcaag caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca
     1081 tggcctgacc ccggaccaag tggtggctat cgccagcaac cacggcggca agcaagcgct
     1141 cgaaacggtg cagcggctgt tgccggtgct gtgccaggac catggcctga ccccggacca
     1201 agtggtggct atcgccagca acattggcgg caagcaagcg ctcgaaacgg tgcagcggct
     1261 gttgccggtg ctgtgccagg accatggcct gaccccggac caagtggtgg ctatcgccag
     1321 caaccacggc ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca
     1381 ggaccatggc ctgactccgg accaagtggt ggctatcgcc agccacgatg gcggcaagca
     1441 agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgacccc
     1501 ggaccaagtg gtggctatcg ccagcaacat tggcggcaag caagcgctcg aaacggtgca
     1561 gcggctgttg ccggtgctgt gccaggacca tggcctgacc ccggaccaag tggtggctat
     1621 cgccagccac gatggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct
     1681 gtgccaggac catggcctga ccccggacca agtggtggct atcgccagca acggtggcgg
     1741 caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct
     1801 gaccccggac caagtggtgg ctatcgccag caaccacggc ggcaagcaag cgctcgaaac
     1861 ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc ctgaccccgg accaagtggt
     1921 ggctatcgcc agcaacattg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc
     1981 ggtgctgtgc caggaccatg gcctgacccc ggaccaagtg gtggctatcg ccagcaacat
     2041 tggcggcaag caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca
     2101 tggcctgacc ccggaccaag tggtggctat cgccagcaac attggcggca agcaagcgct
     2161 cgaaacggtg cagcggctgt tgccggtgct gtgccaggac catggcctga ctccggacca
     2221 agtggtggct atcgccagcc acgatggcgg caagcaagcg ctcgaaacgg tgcagcggct
     2281 gttgccggtg ctgtgccagg accatggcct gaccccggac caagtggtgg ctatcgccag
     2341 caacggtggc ggcaagcaag cgctcgaaag cattgtggcc cagctgagcc ggcctgatcc
     2401 ggcgttggcc gcgttgacca acgaccacct cgtcgccttg gcctgcctcg gcggacgtcc
     2461 tgccatggat gcagtgaaaa agggattgcc gcacgcgccg gaattgatca gaagagtcaa
     2521 tcgccgtatt ggcgaacgca cgtcccatcg cgttgcggga tccatggact acaaggacga
     2581 cgatgacaag agccccaaga agaagagaaa ggtggaggcc agcggttccg gacgggctga
     2641 cgcattggac gattttgatc tggatatgct gggaagtgac gccctcgatg attttgacct
     2701 tgacatgctt ggttcggatg cccttgatga ctttgacctc gacatgctcg gcagtgacgc
     2761 ccttgatgat ttcgacctgg acatgctgat taactctagt tgagaattct gtacagctac
     2821 cgcggccgcg gatctggtta ccactaaacc agcctcaaga acacccgaat ggagtctcta
     2881 agctacataa taccaactta cactttacaa aatgttgtcc cccaaaatgt agccattcgt
     2941 atctgctcct aataaaaaga aagtttcttc acattctaaa aaaaaaaaaa aaaaaaaaaa
     3001 aaaaaaaaaa mccccccccc cccccccctg cagggccaag tcggccggta cccaattcgc
     3061 cctatagtga gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     3121 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     3181 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     3241 ggcaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     3301 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     3361 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     3421 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     3481 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     3541 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     3601 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     3661 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     3721 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3781 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     3841 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     3901 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     3961 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     4021 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     4081 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     4141 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     4201 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     4261 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     4321 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     4381 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     4441 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     4501 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     4561 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     4621 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     4681 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     4741 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     4801 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4861 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4921 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4981 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     5041 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5101 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     5161 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     5221 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     5281 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5341 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5401 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5461 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     5521 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     5581 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     5641 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     5701 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     5761 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     5821 gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc g
//
