LOCUS       sequence-199279-        5517 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for IvT reaction with T3. Encodes LINE-1 specific TALE 
            sequence followed by multi cloning site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5517)
  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 5517)
  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-Tf1-Ctrl
FEATURES             Location/Qualifiers
     source          1..5517
                     /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             2363..2386
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2390..2410
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     3'UTR           2493..2618
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(2706..2725)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2707..2725)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2732..2749)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2732..2748)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2741..2763)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(2890..3345)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3027..3048)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3239..3258
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3372..3476
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3477..3545,3546..4337)
                     /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(3695..3714)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4508..5096
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4997..5016
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5250..5267
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5384..5405
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5420..5425,5426..5443,5444..5450)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5458..5474
                     /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     5463..5485
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5482..5498
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5482..5498
                     /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 ccccggacca agtggtggct atcgccagca acattggcgg
      721 caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct
      781 gactccggac caagtggtgg ctatcgccag ccacgatggc ggcaagcaag cgctcgaaac
      841 ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc ctgaccccgg accaagtggt
      901 ggctatcgcc agcaacattg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc
      961 ggtgctgtgc caggaccatg gcctgacccc ggaccaagtg gtggctatcg ccagcaacca
     1021 cggcggcaag caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca
     1081 tggcctgact ccggaccaag tggtggctat cgccagccac gatggcggca agcaagcgct
     1141 cgaaacggtg cagcggctgt tgccggtgct gtgccaggac catggcctga ccccggacca
     1201 agtggtggct atcgccagca acggtggcgg caagcaagcg ctcgaaacgg tgcagcggct
     1261 gttgccggtg ctgtgccagg accatggcct gaccccggac caagtggtgg ctatcgccag
     1321 caacggtggc ggcaagcaag cgctcgaaac ggtgcagcgg ctgttgccgg tgctgtgcca
     1381 ggaccatggc ctgactccgg accaagtggt ggctatcgcc agccacgatg gcggcaagca
     1441 agcgctcgaa acggtgcagc ggctgttgcc ggtgctgtgc caggaccatg gcctgacccc
     1501 ggaccaagtg gtggctatcg ccagcaacgg tggcggcaag caagcgctcg aaacggtgca
     1561 gcggctgttg ccggtgctgt gccaggacca tggcctgacc ccggaccaag tggtggctat
     1621 cgccagcaac cacggcggca agcaagcgct cgaaacggtg cagcggctgt tgccggtgct
     1681 gtgccaggac catggcctga ccccggacca agtggtggct atcgccagca accacggcgg
     1741 caagcaagcg ctcgaaacgg tgcagcggct gttgccggtg ctgtgccagg accatggcct
     1801 gaccccggac caagtggtgg ctatcgccag caacattggc ggcaagcaag cgctcgaaac
     1861 ggtgcagcgg ctgttgccgg tgctgtgcca ggaccatggc ctgaccccgg accaagtggt
     1921 ggctatcgcc agcaacattg gcggcaagca agcgctcgaa acggtgcagc ggctgttgcc
     1981 ggtgctgtgc caggaccatg gcctgactcc ggaccaagtg gtggctatcg ccagccacga
     2041 tggcggcaag caagcgctcg aaacggtgca gcggctgttg ccggtgctgt gccaggacca
     2101 tggcctgacc ccggaccaag tggtggctat cgccagcaac attggcggca agcaagcgct
     2161 cgaaagcatt gtggcccagc tgagccggcc tgatccggcg ttggccgcgt tgaccaacga
     2221 ccacctcgtc gccttggcct gcctcggcgg acgtcctgcc atggatgcag tgaaaaaggg
     2281 attgccgcac gcgccggaat tgatcagaag agtcaatcgc cgtattggcg aacgcacgtc
     2341 ccatcgcgtt gcgggatcca tggactacaa ggacgacgat gacaagagcc ccaagaagaa
     2401 gagaaaggtg gaggccagcg aattcgatat cggcggccgc gtcgactgat aattctgtac
     2461 agctaccgcg gccgcggatc tggttaccac taaaccagcc tcaagaacac ccgaatggag
     2521 tctctaagct acataatacc aacttacact ttacaaaatg ttgtccccca aaatgtagcc
     2581 attcgtatct gctcctaata aaaagaaagt ttcttcacat tctaaaaaaa aaaaaaaaaa
     2641 aaaaaaaaaa aaaaaamccc cccccccccc cccctgcagg gccaagtcgg ccggtaccca
     2701 attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa cgtcgtgact
     2761 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     2821 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     2881 gcgaatggca aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     2941 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     3001 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     3061 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     3121 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     3181 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     3241 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     3301 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     3361 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3421 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     3481 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     3541 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     3601 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3661 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     3721 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     3781 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3841 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     3901 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     3961 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     4021 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     4081 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     4141 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     4201 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     4261 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     4321 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     4381 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4441 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4501 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4561 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4621 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4681 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4741 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4801 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4861 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4921 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4981 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     5041 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     5101 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     5161 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     5221 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     5281 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     5341 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     5401 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     5461 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagctcg
//
