LOCUS       sequence-105766-        7416 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a 3xFLAG-tagged TAL protein recognizing a telomere 
            repeat..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7416)
  AUTHORS   Fujita T, Asano Y, Ohtsuka J, Takada Y, Saito K, Ohki R, Fujii H
  TITLE     Identification of telomere-associated molecules by engineered 
            DNA-binding molecule-mediated chromatin immunoprecipitation 
            (enChIP).
  JOURNAL   Sci Rep. 2013 Nov 8;3:3171. doi: 10.1038/srep03171.
  PUBMED    24201379
REFERENCE   2  (bases 1 to 7416)
  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: "Sci Rep."; 
            date: "2013-11-8"; volume: "3:3171. doi"; pages: " 
            10.1038/srep03171"
COMMENT     Sequence Label: 3xFN-Tel-TAL/pCMV-7.1
FEATURES             Location/Qualifiers
     source          1..7416
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             931..996
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      1012..1021
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1021..1062
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1075..1095
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3757..4379
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3956..3975)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(4403..4423)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4408..4737
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4588..4723
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4775..4794)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4776..4794)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4808..4824)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4808..4824)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4821..4843)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4832..4848
                     /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(4856..4862,4863..4880,4881..4886))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4901..4922
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5039..5056)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5210..5798)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5290..5309)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5969..6760,6761..6829))
                     /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     6592..6611
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6830..6934)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6961..7389
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7048..7067)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7258..7279
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa
      961 gatcatgaca tcgattacaa ggatgacgat gacaagcttg cggccggccg cgccaccatg
     1021 ggaaaaccta ttcctaatcc tctgctgggc ctggattcta ccggaggcat ggcccctaag
     1081 aaaaagcgga aggtggacgg cggagtggac ctgagaacac tgggatattc tcagcagcag
     1141 caggagaaga tcaagcccaa ggtgagatcc acagtggccc agcaccacga agccctggtg
     1201 ggacacggat ttacacacgc ccacattgtg gccctgtctc agcaccctgc cgccctggga
     1261 acagtggccg tgaaatatca ggatatgatt gccgccctgc ctgaggccac acacgaagcc
     1321 attgtgggag tgggaaaaca gtggtctgga gccagagccc tggaagccct gctgacagtg
     1381 gccggagaac tgagaggacc tcctctgcag ctggatacag gacagctgct gaagattgcc
     1441 aaaaggggcg gagtgaccgc ggtggaagcc gtgcacgcct ggagaaatgc cctgacagga
     1501 gcccctctga acctgacccc cgaacaggtg gtggccattg ccagcaacat cggcggcaag
     1561 caggccctgg aaaccgtgca gagactgctg cccgtgctgt gccaggccca tggcctgaca
     1621 cctgaacagg tggtggctat cgcctctaac aacggaggaa aacaggctct ggaaacagtg
     1681 cagcggctgc tgcctgtgct gtgtcaggct cacggcttga ctccagaaca ggtggtggct
     1741 attgcttcca acaacggggg gaaacaggcc ctggaaactg tgcagcgcct gctgccagtg
     1801 ctgtgccagg ctcacggact gacccccgaa caggtggtgg ccattgccag caacaacggc
     1861 ggcaagcagg ccctggaaac cgtgcagaga ctgctgcccg tgctgtgcca ggcccatggc
     1921 ctgacacctg aacaggtggt ggctatcgcc tctaacggcg gaggaaaaca ggctctggaa
     1981 acagtgcagc ggctgctgcc tgtgctgtgt caggctcacg gcttgactcc agaacaggtg
     2041 gtggctattg cttccaacgg cggggggaaa caggccctgg aaactgtgca gcgcctgctg
     2101 ccagtgctgt gccaggctca cgggctgacc cccgaacagg tggtggccat tgccagcaac
     2161 atcggcggca agcaggccct ggaaaccgtg cagagactgc tgcccgtgct gtgccaggcc
     2221 catggcctga cacctgaaca ggtggtggct atcgcctcta acaacggagg aaaacaggct
     2281 ctggaaacag tgcagcggct gctgcctgtg ctgtgtcagg ctcacggctt gactccagaa
     2341 caggtggtgg ctattgcttc caacaacggg gggaaacagg ccctggaaac tgtgcagcgc
     2401 ctgctgccag tgctgtgcca ggctcacggc ctcactcccg aacaggtggt ggccattgcc
     2461 agcaacaacg gcggcaagca ggccctggaa accgtgcaga gactgctgcc cgtgctgtgc
     2521 caggcccatg gcctgacacc tgaacaggtg gtggctatcg cctctaacgg cggaggaaaa
     2581 caggctctgg aaacagtgca gcggctgctg cctgtgctgt gtcaggctca cggcttgact
     2641 ccagaacagg tggtggctat tgcttccaac ggcgggggga aacaggccct ggaaactgtg
     2701 cagcgcctgc tgccagtgct gtgccaggct cacggactga cccccgaaca ggtggtggcc
     2761 attgccagca acatcggcgg caagcaggcc ctggaaaccg tgcagagact gctgcccgtg
     2821 ctgtgccagg cccatggcct gacacctgaa caggtggtgg ctatcgcctc taacaacgga
     2881 ggaaaacaag cactcgagac agtgcagcgg ctgctgcctg tgctgtgtca ggctcacggc
     2941 ttgactccag aacaggtggt ggctattgct tccaacaacg gggggaaaca ggccctggaa
     3001 actgtgcagc gcctgctgcc agtgctgtgc caggctcacg ggctgacccc cgaacaggtg
     3061 gtggccattg ccagcaacaa cggcggcaag caggccctgg aaaccgtgca gagactgctg
     3121 cccgtgctgt gccaggccca tggcctgaca cctgaacagg tggtggctat cgcctctaac
     3181 ggcggaggaa aacaggctct ggaaacagtg cagcggctgc tgcctgtgct gtgtcaggct
     3241 cacggcttga ctccacagca ggtcgtggca attgctagca acggcggcgg acggcccgcc
     3301 ctggagagca ttgtggccca gctgtctaga cctgatcctg ccctggccgc cctgacaaat
     3361 gatcacctgg tggccctggc ctgtctggga ggcagacctg ccctggatgc cgtgaaaaaa
     3421 ggactgcctc acgcccctgc cctgatcaag agaacaaata gaagaatccc cgagcggacc
     3481 tctcacagag tggccgatca cgcccaggtg gtgagagtgc tgggattttt tcagtgtcac
     3541 tctcaccctg cccaggcctt tgatgatgcc atgacacagt ttggcatgag cagacacgga
     3601 ctgctgcagc tgtttagaag agtgggagtg acagaactgg aggccagatc cggaaccctg
     3661 cctcctgcct ctcagagatg ggataggatt ctgcagggtt cccgtttaat tcatcgatag
     3721 atctgatatc ggtaccagtc gactctagag gatcccgggt ggcatccctg tgacccctcc
     3781 ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata
     3841 aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg
     3901 ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg cggggtctat
     3961 tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc cgcctcctgg
     4021 gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc atgcatgacc
     4081 aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt ggccaggctg
     4141 gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta
     4201 caggcgtgaa ccactgctcc cttccctgtc cttctgattt taaaataact ataccagcag
     4261 gaggacgtcc agacacagca taggctacct ggccatgccc aaccggtggg acatttgagt
     4321 tgcttgcttg gcactgtcct ctcatgcgtt gggtccactc agtagatgcc tgttgaattg
     4381 ggtacgcggc cagcttggct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4441 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     4501 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4561 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4621 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     4681 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     4741 ctcctcgagg aactgaaaaa ccagaaagtt aattccctat agtgagtcgt attaaattcg
     4801 taatcatgtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4861 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4921 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4981 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5041 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5101 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5161 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5221 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5281 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5341 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5401 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5461 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5521 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5581 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5641 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5701 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5761 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5821 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5881 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5941 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6001 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6061 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6121 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6181 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6241 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6301 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6361 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6421 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6481 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6541 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6601 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6661 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6721 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6781 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6841 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6901 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     6961 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     7021 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     7081 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     7141 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     7201 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     7261 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     7321 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     7381 acgcgaattt taacaaaata ttaacgctta caattt
//
