LOCUS       sequence-243409-        8737 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express M.EcoGII fused to TRF1 with detabilization domain - 
            inducible.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8737)
  AUTHORS   Sobecki M, Souaid C, Boulay J, Guerineau V, Noordermeer D, Crabbe L
  TITLE     MadID, a Versatile Approach to Map Protein-DNA Interactions, 
            Highlights Telomere-Nuclear Envelope Contact Sites in Human Cells.
  JOURNAL   Cell Rep. 2018 Dec 4;25(10):2891-2903.e5. doi: 
            10.1016/j.celrep.2018.11.027.
  PUBMED    30517874
REFERENCE   2  (bases 1 to 8737)
  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"; doi: 
            "10.1016/j.celrep.2018.11"; journalName: "Cell Rep"; date: 
            "2018-12-4- 4"; volume: "25"; issue: "10"; pages: "2891-2903.e5"
COMMENT     Sequence Label: pRetroX-PTuner DD-M.EcoGII-v5-Telomeric
            repeat...
FEATURES             Location/Qualifiers
     source          1..8737
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     37..59
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     51..68
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     52..68
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             201..670
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    735..1092
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     intron          1155..1358
                     /label=EF-1-alpha intron A
                     /note="truncated intron upstream of the start codon of 
                     human EF-1-alpha"
     primer_bind     1315..1335
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     exon            1359..1388
                     /label=EF-1-alpha exon
                     /note="truncated exon 2 from human EF-1-alpha, beginning 
                     with a GT-to-CC mutation that reduces splicing efficiency"
     CDS             join(1448..1450,1451..1771)
                     /codon_start=1
                     /product="destabilization domain that can be stabilized by 
                     Shield1 in the ProteoTuner(TM) system"
                     /label=DD
                     /note="L106P mutant of FKBP12"
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKPE"
     CDS             2939..2980
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    join(4318..4892,4893..4895,4896..4904)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4485..4502)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4712..4731
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4924..5520
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4924..4943)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5420..5440
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5580..6173
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6189..6205)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6189..6205)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6202..6224)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6213..6229
                     /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(6237..6243,6244..6261,6262..6267))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6282..6303
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6420..6437)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6591..7179)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6671..6690)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7350..8141,8142..8210))
                     /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"
     promoter        complement(8211..8315)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8383..8401
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8439..8461)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8561..8580
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
       61 cggccagtgc caagcttatg tgaaagaccc cacctgtagg tttggcaagc tagcttaagt
      121 aacgccattt tgcaaggcat ggaaaaatac ataactgaga atagaaaagt tcagatcaag
      181 gtcaggaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc
      241 ctgccccggc tcagggccaa gaacagatgg aacagctgaa tatgggccaa acaggatatc
      301 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc
      361 cagccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa
      421 atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc
      481 ttctgctccc cgggctcaat aaaagagccc acaacccctc actcggcgcg ccagtcctcc
      541 gattgactga gtcgcccggg tacccgtgta tccaataaac cctcttgcag ttgcatccga
      601 cttgtggcct cgctgttcct tgggagggtc tcctctgagt gattgactac ccgtcagcgg
      661 gggtctttca tttgggggct cgtccgggat cgggagaccc ctgcccaggg accaccgacc
      721 caccaccggg aggtaagctg gccagcaact tatctgtgtc tgtccgattg tctagtgtct
      781 atgactgatt ttatgcgcct gcgtcggtac tagttagcta actagctctg tatctggcgg
      841 acccgtggtg gaactgacga gttcggaaca cccggccgca accctgggag acgtcccagg
      901 gacttcgggg gccgtttttg tggcccgacc tgagtccaaa aatcccgatc gttttggact
      961 ctttggtgca ccccccttag aggagggata tgtggttctg gtaggagacg agaacctaaa
     1021 acagttcccg cctccgtctg aatttttgct ttcggtttgg gaccgaagcc gcgccgcgcg
     1081 tcttgtctgc tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc
     1141 tgaaaatacg cgtgtcgagc ttttggagta cgtcgtcttt aggttggggg gaggggtttt
     1201 atgcgatgga gtttccccac actgagtggg tggagactga agttaggcca gcttggcact
     1261 tgatgtaatt ctccttggaa tttgcccttt ttgagtttgg atcttggttc attctcaagc
     1321 ctcagacagt ggttcaaagt ttttttcttc catttcagcc gtcgtgaaaa ctacccctaa
     1381 aagccaaaaa aatcgaattc ccaaacttaa gcttggtacc gagctcggat ctcacgtggg
     1441 cccgcggatg ggagtgcagg tggaaaccat ctccccagga gacgggcgca ccttccccaa
     1501 gcgcggccag acctgtgtgg tgcactacac cgggatgctt gaagatggaa agaaagtcga
     1561 ttcctcccgg gacagaaaca agccctttaa gtttatgcta ggcaagcagg aggtgatccg
     1621 aggctgggaa gaaggggttg cccagatgag tgtgggtcag agagccaaac tgactatatc
     1681 tccagattat gcctatggtg ccactgggca cccaggcatc atcccaccac atgccactct
     1741 cgtcttcgat gtggagcttc taaaaccgga accgcggccg cagatctatc gatggatcct
     1801 atccggactc agatctcgag ctcaagcttc gaattctgca gtcgacggta ccgcgggcag
     1861 cgctgccaac ctgccgggga tccgtatgct taatactgta aaaatatcca gttgtgagtt
     1921 aatcaacgcc gactgcctgg aatttatccg gtcgttaccc gaaaattctg ttgacctgat
     1981 agtcacggac ccgccgtact ttaaagtgaa gcccgagggc tgggataacc agtggaaggg
     2041 cgacgatgat tacctgaagt ggctggacca gtgtctggcg cagttctggc gggtgctgaa
     2101 acctgccgga agtctttacc tgttctgtgg ccatcgcctg gcatctgaca ttgaaatcat
     2161 gatgcgtgaa cgcttcagtg tgctgaacca tattatctgg gcaaagccgt ccggacgctg
     2221 gaacgggtgc aacaaggaaa gcctgcgggc gtatttcccc gccacagagc gcattctgtt
     2281 cgcggaacat tatcaggggc cgtatcgccc gaaagatgcc gggtatgagg cgaagggcag
     2341 ggcactgaaa cagcatgtga tggccccgct gattgcttac tttcgtgatg cgcgcgctgc
     2401 cctggggata acggcaaaac agattgcaga tgccacagga aagaaaaaca tggtgccgca
     2461 ctggttcagt gccagtcagt ggcagctacc gaacgaaagc gattatctga aattacagtc
     2521 gctgtttgcc cgggtggcag aagagaaaca tcagcgcggg gaactggaaa agccacacca
     2581 ccagctggtc agcacataca gtgagctgaa ccggaagtat atggaactgc tgagtgaata
     2641 taaaaatttg cggcggtatt tcggtgtgac ggtgcaggtg ccgtacaccg atgtgtggac
     2701 gtataaaccg gtgcagtact atccagggaa acatccgtgc gaaaaaccgg cagaaatgct
     2761 gcagcagata atcagcgcaa gtagtcgtcc tggtgatctg gttgcggatt ttttcatggg
     2821 gtcgggttca acggtaaaag cggcgatggc actggggcgt cgtgcgattg gtgttgagct
     2881 ggagaccgga cgttttgagc agacagtcag ggaagttcag gatttaatcg ttccatgggg
     2941 taagcctatc cctaaccctc tcctcggtct cgattctacg cgtaccggca tggcggagga
     3001 tgtttcctca gcggccccga gcccgcgggg ctgtgcggat ggtagggatg ccgaccctac
     3061 tgaggagcag atggcagaaa cagagagaaa cgacgaggag cagttcgaat gccaggaact
     3121 gctcgagtgc caggtgcagg tgggggcccc cgaggaggag gaggaggagg aggaggacgc
     3181 gggcctggtg gccgaggccg aggccgtggc tgccggctgg atgctcgatt tcctctgcct
     3241 ctctctttgc cgagctttcc gcgacggccg ctccgaggac ttccgcagga cccgcaacag
     3301 cgcagaggct attattcatg gactatccag tctaacagct tgccagttga gaacgatata
     3361 catatgtcag tttttgacaa gaattgcagc aggaaaaacc cttgatgcac agtttgaaaa
     3421 tgatgaacga attacaccct tggaatcagc cctgatgatt tggggttcaa ttgaaaagga
     3481 acatgacaaa cttcatgaag aaatacagaa tttaattaaa attcaggcta tagctgtttg
     3541 tatggaaaat ggcaacttta aagaagcaga agaagtcttt gaaagaatat ttggtgatcc
     3601 aaattctcat atgcctttca aaagcaaatt gcttatgata atctctcaga aagatacatt
     3661 tcattccttt tttcaacact tcagctacaa ccacatgatg gagaaaatta agagttatgt
     3721 gaattatgtg ctaagtgaaa aatcatcaac ctttctaatg aaggcagcgg caaaagtagt
     3781 agaaagcaaa aggacaagaa caataacttc tcaagataaa cctagtggta atgatgttga
     3841 aatggaaact gaagctaatt tggatacaag aaaaagtgtt agtgacaaac agtctgcggt
     3901 aactgaatcc tcagagggta cagtatcctt attgaggtct cacaagaatc ttttcttatc
     3961 taagttgcaa catggaaccc agcaacaaga ccttaataag aaagaaagaa gagtaggaac
     4021 tcctcaaagt acaaaaaaga aaaaagaaag cagaagagcc actgaaagca gaatacctgt
     4081 ttcaaagagt cagccggtaa ctcctgaaaa acatcgagct agaaaaagac aggcatggct
     4141 ttgggaagaa gacaagaatt tgagatctgg cgtgaggaaa tatggagagg gaaactggtc
     4201 taaaatactg ttgcattata aattcaacaa ccggacaagt gtcatgttaa aagacagatg
     4261 gaggaccatg aagaaactaa aactgatttc ctcagacagc gaagactgac catggcgccc
     4321 ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc
     4381 gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa
     4441 acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat
     4501 gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac
     4561 aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg
     4621 cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt
     4681 tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg
     4741 gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac
     4801 atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga
     4861 cgtggttttc ctttgaaaaa cacgatgata agcttgccac aacccacaag gagacgacct
     4921 tccatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc
     4981 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgacccg
     5041 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
     5101 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
     5161 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
     5221 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
     5281 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg
     5341 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
     5401 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
     5461 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
     5521 accggtgcgg cttcgaggat aaaataaaag attttattta gtctccagaa aaagggggga
     5581 atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca
     5641 tggaaaaata cataactgag aatagagaag ttcagatcaa ggtcaggaac agatggaaca
     5701 gctgaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
     5761 agaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc
     5821 cccggctcag ggccaagaac agatggtccc cagatgcggt ccagccctca gcagtttcta
     5881 gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt
     5941 gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa
     6001 taaaagagcc cacaacccct cactcggggc gccagtcctc cgattgactg agtcgcccgg
     6061 gtacccgtgt atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc
     6121 ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtctttc attgaattcg
     6181 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     6241 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     6301 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     6361 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6421 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6481 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6541 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6601 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6661 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6721 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6781 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6841 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6901 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6961 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     7021 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     7081 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     7141 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     7201 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     7261 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     7321 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     7381 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     7441 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     7501 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     7561 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     7621 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     7681 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     7741 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     7801 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     7861 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     7921 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     7981 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     8041 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     8101 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     8161 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     8221 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     8281 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     8341 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     8401 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     8461 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     8521 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     8581 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     8641 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     8701 tcttcgctat tacgccagct ggcgaaaggg ggatgtg
//
