LOCUS       sequence-72268-e        9014 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the restriction enzyme IPpoI under the control of Shield1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9014)
  AUTHORS   Goldstein M, Derheimer FA, Tait-Mulder J, Kastan MB
  TITLE     Nucleolin mediates nucleosome disruption critical for DNA 
            double-strand break repair.
  JOURNAL   Proc Natl Acad Sci U S A. 2013 Sep 30.
  PUBMED    24082117
REFERENCE   2  (bases 1 to 9014)
  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: "Proc Natl 
            Acad Sci U S A. 2013 Sep 30."
COMMENT     Sequence Label: pCL20C-ddIPpoI
FEATURES             Location/Qualifiers
     source          1..9014
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     LTR             385..642
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    689..814
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1306..1423
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    1783..2016
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2201..2245
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     LTR             2498..3014
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    3078..3419
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             3486..3902
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     3832..3854
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     3870..3886
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             join(3952..3954,3955..4275)
                     /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             4291..4317
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(6036..6055)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6101..6156
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6155..6174)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6517..6533)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6517..6533)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6530..6552)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6541..6557
                     /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(6565..6571,6572..6589,6590..6595))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6610..6631
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6690..6885
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6736..6871
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6891..7025
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6928..6947)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6982..7001
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7093..7110)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7264..7852)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7344..7363)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8023..8814,8815..8883))
                     /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     8646..8665
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8884..8988)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggaggcg tggcctgggc gggactgggg agtggcgagc
      421 cctcagatcc tgcatataag cagctgcttt ttgcctgtac tgggtctctc tggttagacc
      481 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
      541 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
      601 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga
      661 cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc
      721 gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg
      781 aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc
      841 gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat
      901 agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc
      961 agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga
     1021 acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat
     1081 aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaaaaa
     1141 agcacagcaa gcagcaggat cttcagacct ggaaattccc tacaatcccc aaagtcaagg
     1201 agtagtagaa tctatgaata aagaattaaa gaaaattata ggacaggtaa gagatcaggc
     1261 tgaacatctt aagacagcag tacaaatggc agtattcatc cacaatttta aaagaaaagg
     1321 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     1381 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     1441 cagcagaaat ccactttgga aaggaccagc aaagctcctc tggaaaggtg aaggggcagt
     1501 agtaatacaa gataatagtg acataaaagt agtgccaaga agaaaagcaa agatcattag
     1561 ggattatgga aaacagatgg caggtgatga ttgtgtggca agtagacagg atgaggatta
     1621 gaacatggaa aagtttagta aaacaccata aggaggagat atgagggaca attggagaag
     1681 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1741 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1801 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1861 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1921 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1981 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     2041 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     2101 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     2161 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     2221 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     2281 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     2341 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     2401 cccaaccccg aggggaccga gctcaagctt cgaacgcgtt aacgggccca gcttcgataa
     2461 aataaaagat tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt
     2521 ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaatacat aactgagaat
     2581 agagaagttc agatcaaggt taggaacaga gagacagcag aatatgggcc aaacaggata
     2641 tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg
     2701 tcccgccctc agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg
     2761 aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc
     2821 gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcggcg cgccagtcct
     2881 ccgatagact gcgtcgcccg ggtacccgta ttcccaataa agcctcttgc tgtttgcatc
     2941 cgaatcgtgg actcgctgat ccttgggagg gtctcctcag attgattgac tgcccacctc
     3001 gggggtcttt catttggagg ttccaccgag atttggagac ccctgcccag ggaccaccga
     3061 cccccccgcc gggaggtaag ctggccagcg gtcgtttcgt gtctgtctct gtctttgtgc
     3121 gtgtttgtgc cggcatctaa tgtttgcgcc tgcgtctgta ctagttagct aactagctct
     3181 gtatctggcg gacccgtggt ggaactgacg agttctgaac acccggccgc aaccctggga
     3241 gacgtcccag ggactttggg ggccgttttt gtggcccgac ctgaggaagg gagtcgatgt
     3301 ggaatccgac cccgtcagga tatgtggttc tggtaggaga cgagaaccta aaacagttcc
     3361 cgcctccgtc tgaatttttg ctttcggttt ggaaccgaag ccgcgcgtct tgtctgctgc
     3421 agcgctgcag catcgttctg tgttgtctct gtctgactgt gtttctgtat ttgtctgaaa
     3481 attagggcca gactgttacc actcccttaa gtttgacctt aggtcactgg aaagatgtcg
     3541 agcggatcgc tcacaaccag tcggtagatg tcaagaagag acgttgggtt accttctgct
     3601 ctgcagaatg gccaaccttt aacgtcggat ggccgcgaga cggcaccttt aaccgagacc
     3661 tcatcaccca ggttaagatc aaggtctttt cacctggccc gcatggacac ccagaccagg
     3721 tcccctacat cgtgacctgg gaagccttgg cttttgaccc ccctccctgg gtcaagccct
     3781 ttgtacaccc taagcctccg cctcctcttc ctccatccgc cccgtctctc ccccttgaac
     3841 ctcctcgttc gaccccgcct cgatcctccc tttatccagc cctcactcct tctctaggcg
     3901 ccggaattct gcagtcgacg gtaccgcggg cccgggatcc accggtggat catgggagtg
     3961 caggtggaga ctatctcccc aggagacggg cgcaccttcc ccaagcgcgg ccagacctgc
     4021 gtggtgcact acaccgggat gcttgaagat ggaaagaaag ttgattcctc ccgggacaga
     4081 aacaagccct ttaagtttat gctaggcaag caggaggtga tccgaggctg ggaagaaggg
     4141 gttgcccaga tgagtgtggg tcagagagcc aaactgacta tatctccaga ttatgcctat
     4201 ggtgccactg ggcacccagg catcatccca ccacatgcca ctctcgtctt cgatgtggag
     4261 cttctaaaac cggaagaatt caccatggca tacccatacg atgttccaga ttacgctagc
     4321 ggatctcgaa atgaaatggg tgcttcagga gacatgaggg ctgccaacct ttggccaagc
     4381 cctcttgtga ttaagcacac taagaagaat agccctgcct tgtccttgac agctgaccag
     4441 atggtcagtg ccttgttgga tgctgaaccg cccatgatct attctgaata tgatccttct
     4501 agacccttca gtgaagcctc aatgatgggc ttattgacca acctagcaga tagggagctg
     4561 gttcatatga tcaactgggc aaagagagtg ccaggctttg gggacttgaa tctccatgat
     4621 caggtccacc ttctcgagtg tgcctggctg gagattctga tgattggtct cgtctggcgc
     4681 tccttggaac acccggggaa gctcctgttt gctcctaact tgctcctgga caggaatcaa
     4741 ggtaaatgtg tggaaggcat ggtggagatc tttgacatgt tgcttgctac gtcaagtcgg
     4801 ttccgcatga tgaacctgca gggtgaagag tttgtgtgcc tcaaatccat cattttgctt
     4861 aattccggag tgtacacgtt tctgtccagc accttgaagt ctctggaaga gaaggaccac
     4921 atccaccgtg tcctggacaa gatcacagac actttgatcc acctgatggc caaagctggc
     4981 ctgactctgc agcagcagca tcgccgccta gctcagctcc ttctcattct ttcccatatc
     5041 cggcatatga gtaacaaacg catggagcat ctctacaaca tgaaatgcaa gaacgtggta
     5101 cccctctatg acctgctcct ggagatgttg gatgcccacc gcctccatgc cccagccagt
     5161 cgcatgggag tgcccccaga ggagcccagc cagacccagc tggccaccac cagctccact
     5221 tcagcacatt ccttacaaac ctactacata cccccggaag cagagggctt ccccaacacg
     5281 atcggatcta tggcgctcac caatgctcaa atcttggctg tgattgacag ctgggaagaa
     5341 acagtcggtc agtttccagt gataacgcac catgtaccat taggtggcgg tctgcaagga
     5401 acgctccatt gttacgagat ccccctagca gctccttatg gggttggctt tgctaagaat
     5461 gggcctaccc gctggcaata caaacggaca atcaatcaag tcgtccacag atggggatcc
     5521 cacacagtcc cctttctatt agaaccggat aacatcaacg gcaaaacctg cacagcatcg
     5581 cacctatgtc ataatactcg atgccacaat cccttgcact tgtgctggga gtcactagac
     5641 gacaacaaag gcagaaactg gtgcccgggt cccaacgggg gatgtgtcca tgcggtggtt
     5701 tgtttaaggc agggtccgtt gtacggcccg ggggcgactg tggcaggtcc tcaacaaagg
     5761 ggcagtcact ttgtggtata agtcggcggc cgcatcgatg ccgtatacgg tacctttaag
     5821 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact
     5881 ggaagggcta attcactccc aaagaagaca agatctgctt tttgcctgta ctgggtctct
     5941 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
     6001 gcctcaataa agcttcagct gctcgagcta gcagatcttt ttccctctgc caaaaattat
     6061 ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc
     6121 attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca
     6181 aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg
     6241 ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct
     6301 gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt
     6361 tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc
     6421 agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga
     6481 tccctcgacc tgcagcccaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     6541 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     6601 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     6661 gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt agtcagcaac catagtcccg
     6721 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     6781 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     6841 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct aacttgttta
     6901 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     6961 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     7021 ggatccgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     7081 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     7141 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     7201 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     7261 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     7321 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     7381 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     7441 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     7501 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     7561 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     7621 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     7681 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     7741 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     7801 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     7861 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     7921 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     7981 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     8041 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     8101 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     8161 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     8221 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     8281 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     8341 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     8401 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     8461 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     8521 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     8581 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     8641 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     8701 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     8761 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     8821 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     8881 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     8941 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     9001 aaaagtgcca cctg
//
