LOCUS       sequence-238098-        7414 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Express M.EcoGII with detabilization domain - inducible.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7414)
  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 7414)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-linker-M.EcoGII
FEATURES             Location/Qualifiers
     source          1..7414
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          78..281
                     /label=EF-1-alpha intron A
                     /note="truncated intron upstream of the start codon of 
                     human EF-1-alpha"
     primer_bind     238..258
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     exon            282..311
                     /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(371..373,374..694)
                     /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             809..850
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    join(1918..2492,2493..2495,2496..2504)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2085..2102)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2312..2331
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2524..3120
                     /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(2524..2543)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3020..3040
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3180..3773
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3789..3805)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3789..3805)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3802..3824)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3813..3829
                     /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(3837..3843,3844..3861,3862..3867))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3882..3903
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4020..4037)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4191..4779)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4271..4290)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4950..5741,5742..5810))
                     /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(5811..5915)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5983..6001
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6039..6061)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6161..6180
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6374..6396
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6388..6405
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6389..6405
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             6538..7007
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    join(7072..7414,1..15)
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
ORIGIN
        1 gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt
       61 gtctgaaaat acgcgtgtcg agcttttgga gtacgtcgtc tttaggttgg ggggaggggt
      121 tttatgcgat ggagtttccc cacactgagt gggtggagac tgaagttagg ccagcttggc
      181 acttgatgta attctccttg gaatttgccc tttttgagtt tggatcttgg ttcattctca
      241 agcctcagac agtggttcaa agtttttttc ttccatttca gccgtcgtga aaactacccc
      301 taaaagccaa aaaaatcgaa ttcccaaact taagcttggt accgagctcg gatctcacgt
      361 gggcccgcgg atgggagtgc aggtggaaac catctcccca ggagacgggc gcaccttccc
      421 caagcgcggc cagacctgtg tggtgcacta caccgggatg cttgaagatg gaaagaaagt
      481 cgattcctcc cgggacagaa acaagccctt taagtttatg ctaggcaagc aggaggtgat
      541 ccgaggctgg gaagaagggg ttgcccagat gagtgtgggt cagagagcca aactgactat
      601 atctccagat tatgcctatg gtgccactgg gcacccaggc atcatcccac cacatgccac
      661 tctcgtcttc gatgtggagc ttctaaaacc ggaaccgcgg ccgctatccg gactcagatc
      721 tcgagctcaa gcttcgaatt ctgcagtcga cggtaccgcg ggcagcgctg ccaacctgcc
      781 cggcggccgc agatctatcg atggatccgg taagcctatc cctaaccctc tcctcggtct
      841 cgattctacg cgtaccggca tgcttaatac tgtaaaaata tccagttgtg agttaatcaa
      901 cgccgactgc ctggaattta tccggtcgtt acccgaaaat tctgttgacc tgatagtcac
      961 ggacccgccg tactttaaag tgaagcccga gggctgggat aaccagtgga agggcgacga
     1021 tgattacctg aagtggctgg accagtgtct ggcgcagttc tggcgggtgc tgaaacctgc
     1081 cggaagtctt tacctgttct gtggccatcg cctggcatct gacattgaaa tcatgatgcg
     1141 tgaacgcttc agtgtgctga accatattat ctgggcaaag ccgtccggac gctggaacgg
     1201 gtgcaacaag gaaagcctgc gggcgtattt ccccgccaca gagcgcattc tgttcgcgga
     1261 acattatcag gggccgtatc gcccgaaaga tgccgggtat gaggcgaagg gcagggcact
     1321 gaaacagcat gtgatggccc cgctgattgc ttactttcgt gatgcgcgcg ctgccctggg
     1381 gataacggca aaacagattg cagatgccac aggaaagaaa aacatggtgc cgcactggtt
     1441 cagtgccagt cagtggcagc taccgaacga aagcgattat ctgaaattac agtcgctgtt
     1501 tgcccgggtg gcagaagaga aacatcagcg cggggaactg gaaaagccac accaccagct
     1561 ggtcagcaca tacagtgagc tgaaccggaa gtatatggaa ctgctgagtg aatataaaaa
     1621 tttgcggcgg tatttcggtg tgacggtgca ggtgccgtac accgatgtgt ggacgtataa
     1681 accggtgcag tactatccag ggaaacatcc gtgcgaaaaa ccggcagaaa tgctgcagca
     1741 gataatcagc gcaagtagtc gtcctggtga tctggttgcg gattttttca tggggtcggg
     1801 ttcaacggta aaagcggcga tggcactggg gcgtcgtgcg attggtgttg agctggagac
     1861 cggacgtttt gagcagacag tcagggaagt tcaggattta atcgtttaac catggcgccc
     1921 ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc
     1981 gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa
     2041 acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat
     2101 gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac
     2161 aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg
     2221 cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt
     2281 tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg
     2341 gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac
     2401 atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga
     2461 cgtggttttc ctttgaaaaa cacgatgata agcttgccac aacccacaag gagacgacct
     2521 tccatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc
     2581 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgacccg
     2641 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
     2701 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
     2761 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
     2821 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
     2881 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg
     2941 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
     3001 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
     3061 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
     3121 accggtgcgg cttcgaggat aaaataaaag attttattta gtctccagaa aaagggggga
     3181 atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca
     3241 tggaaaaata cataactgag aatagagaag ttcagatcaa ggtcaggaac agatggaaca
     3301 gctgaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
     3361 agaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc
     3421 cccggctcag ggccaagaac agatggtccc cagatgcggt ccagccctca gcagtttcta
     3481 gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt
     3541 gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa
     3601 taaaagagcc cacaacccct cactcggggc gccagtcctc cgattgactg agtcgcccgg
     3661 gtacccgtgt atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc
     3721 ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtctttc attgaattcg
     3781 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     3841 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     3901 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     3961 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4021 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4081 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4141 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4201 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4261 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4321 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4381 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4441 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4501 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4561 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4621 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4681 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4741 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4801 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4861 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4921 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4981 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5041 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5101 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5161 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5221 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5281 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5341 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5401 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5461 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5521 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     5581 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     5641 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5701 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5761 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     5821 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     5881 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     5941 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     6001 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     6061 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     6121 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     6181 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     6241 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     6301 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     6361 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gcttatgtga
     6421 aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga
     6481 aaaatacata actgagaata gaaaagttca gatcaaggtc aggaacagat ggaacagctg
     6541 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     6601 cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
     6661 gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga
     6721 accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac
     6781 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccgg gctcaataaa
     6841 agagcccaca acccctcact cggcgcgcca gtcctccgat tgactgagtc gcccgggtac
     6901 ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggcctcgc tgttccttgg
     6961 gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt gggggctcgt
     7021 ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg taagctggcc
     7081 agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta tgcgcctgcg
     7141 tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt
     7201 cggaacaccc ggccgcaacc ctgggagacg tcccagggac ttcgggggcc gtttttgtgg
     7261 cccgacctga gtccaaaaat cccgatcgtt ttggactctt tggtgcaccc cccttagagg
     7321 agggatatgt ggttctggta ggagacgaga acctaaaaca gttcccgcct ccgtctgaat
     7381 ttttgctttc ggtttgggac cgaagccgcg ccgc
//
