LOCUS       sequence-333157-        9015 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 9015)
  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 9015)
  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..9015
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     LTR             382..639
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    686..811
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1420
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    1780..2013
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2198..2242
                     /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             2495..3011
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    3075..3416
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             3483..3899
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     3829..3851
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     3867..3883
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             join(3949..3951,3952..4272)
                     /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             4288..4314
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(6033..6052)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6098..6153
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6152..6171)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6514..6530)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6514..6530)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6527..6549)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6538..6554
                     /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(6562..6568,6569..6586,6587..6592))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6607..6628
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6687..6882
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6733..6868
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6888..7022
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6925..6944)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6979..6998
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7090..7107)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7261..7849)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7341..7360)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8020..8811,8812..8880))
                     /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     8643..8662
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8881..8985)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg ggaggcgtgg cctgggcggg actggggagt ggcgagccct
      421 cagatcctgc atataagcag ctgctttttg cctgtactgg gtctctctgg ttagaccaga
      481 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
      541 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
      601 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt
      661 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg
      721 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
      781 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg
      841 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt
      901 atgggcaagc agggagctag aacgattcgc agttaatact ggcctgttag aaacatcaga
      961 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact
     1021 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa
     1081 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaaaaaagc
     1141 acagcaagca gcaggatctt cagacctgga aattccctac aatccccaaa gtcaaggagt
     1201 agtagaatct atgaataaag aattaaagaa aattatagga caggtaagag atcaggctga
     1261 acatcttaag acagcagtac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     1321 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     1381 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     1441 cagaaatcca ctttggaaag gaccagcaaa gctcctctgg aaaggtgaag gggcagtagt
     1501 aatacaagat aatagtgaca taaaagtagt gccaagaaga aaagcaaaga tcattaggga
     1561 ttatggaaaa cagatggcag gtgatgattg tgtggcaagt agacaggatg aggattagaa
     1621 catggaaaag tttagtaaaa caccataagg aggagatatg agggacaatt ggagaagtga
     1681 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
     1741 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
     1801 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag
     1861 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
     1921 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc
     1981 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
     2041 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
     2101 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat
     2161 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga
     2221 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat
     2281 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
     2341 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc
     2401 aaccccgagg ggaccgagct caagcttcga acgcgttaac gggcccagct tcgataaaat
     2461 aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg
     2521 gcaagctagc ttaagtaacg ccattttgca aggcatggaa aatacataac tgagaataga
     2581 gaagttcaga tcaaggttag gaacagagag acagcagaat atgggccaaa caggatatct
     2641 gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc
     2701 cgccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa
     2761 tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct
     2821 tctgctcccc gagctcaata aaagagccca caacccctca ctcggcgcgc cagtcctccg
     2881 atagactgcg tcgcccgggt acccgtattc ccaataaagc ctcttgctgt ttgcatccga
     2941 atcgtggact cgctgatcct tgggagggtc tcctcagatt gattgactgc ccacctcggg
     3001 ggtctttcat ttggaggttc caccgagatt tggagacccc tgcccaggga ccaccgaccc
     3061 ccccgccggg aggtaagctg gccagcggtc gtttcgtgtc tgtctctgtc tttgtgcgtg
     3121 tttgtgccgg catctaatgt ttgcgcctgc gtctgtacta gttagctaac tagctctgta
     3181 tctggcggac ccgtggtgga actgacgagt tctgaacacc cggccgcaac cctgggagac
     3241 gtcccaggga ctttgggggc cgtttttgtg gcccgacctg aggaagggag tcgatgtgga
     3301 atccgacccc gtcaggatat gtggttctgg taggagacga gaacctaaaa cagttcccgc
     3361 ctccgtctga atttttgctt tcggtttgga accgaagccg cgcgtcttgt ctgctgcagc
     3421 gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg tctgaaaatt
     3481 agggccagac tgttaccact cccttaagtt tgaccttagg tcactggaaa gatgtcgagc
     3541 ggatcgctca caaccagtcg gtagatgtca agaagagacg ttgggttacc ttctgctctg
     3601 cagaatggcc aacctttaac gtcggatggc cgcgagacgg cacctttaac cgagacctca
     3661 tcacccaggt taagatcaag gtcttttcac ctggcccgca tggacaccca gaccaggtcc
     3721 cctacatcgt gacctgggaa gccttggctt ttgacccccc tccctgggtc aagccctttg
     3781 tacaccctaa gcctccgcct cctcttcctc catccgcccc gtctctcccc cttgaacctc
     3841 ctcgttcgac cccgcctcga tcctcccttt atccagccct cactccttct ctaggcgccg
     3901 gaattctgca gtcgacggta ccgcgggccc gggatccacc ggtggatcat gggagtgcag
     3961 gtggagacta tctccccagg agacgggcgc accttcccca agcgcggcca gacctgcgtg
     4021 gtgcactaca ccgggatgct tgaagatgga aagaaagttg attcctcccg ggacagaaac
     4081 aagcccttta agtttatgct aggcaagcag gaggtgatcc gaggctggga agaaggggtt
     4141 gcccagatga gtgtgggtca gagagccaaa ctgactatat ctccagatta tgcctatggt
     4201 gccactgggc acccaggcat catcccacca catgccactc tcgtcttcga tgtggagctt
     4261 ctaaaaccgg aagaattcac catggcatac ccatacgatg ttccagatta cgctagcgga
     4321 tctcgaaatg aaatgggtgc ttcaggagac atgagggctg ccaacctttg gccaagccct
     4381 cttgtgatta agcacactaa gaagaatagc cctgccttgt ccttgacagc tgaccagatg
     4441 gtcagtgcct tgttggatgc tgaaccgccc atgatctatt ctgaatatga tccttctaga
     4501 cccttcagtg aagcctcaat gatgggctta ttgaccaacc tagcagatag ggagctggtt
     4561 catatgatca actgggcaaa gagagtgcca ggctttgggg acttgaatct ccatgatcag
     4621 gtccaccttc tcgagtgtgc ctggctggag attctgatga ttggtctcgt ctggcgctcc
     4681 ttggaacacc cggggaagct cctgtttgct cctaacttgc tcctggacag gaatcaaggt
     4741 aaatgtgtgg aaggcatggt ggagatcttt gacatgttgc ttgctacgtc aagtcggttc
     4801 cgcatgatga acctgcaggg tgaagagttt gtgtgcctca aatccatcat tttgcttaat
     4861 tccggagtgt acacgtttct gtccagcacc ttgaagtctc tggaagagaa ggaccacatc
     4921 caccgtgtcc tggacaagat cacagacact ttgatccacc tgatggccaa agctggcctg
     4981 actctgcagc agcagcatcg ccgcctagct cagctccttc tcattctttc ccatatccgg
     5041 catatgagta acaaacgcat ggagcatctc tacaacatga aatgcaagaa cgtggtaccc
     5101 ctctatgacc tgctcctgga gatgttggat gcccaccgcc tccatgcccc agccagtcgc
     5161 atgggagtgc ccccagagga gcccagccag acccagctgg ccaccaccag ctccacttca
     5221 gcacattcct tacaaaccta ctacataccc ccggaagcag agggcttccc caacacgatc
     5281 ggatctatgg cgctcaccaa tgctcaaatc ttggctgtga ttgacagctg ggaagaaaca
     5341 gtcggtcagt ttccagtgat aacgcaccat gtaccattag gtggcggtct gcaaggaacg
     5401 ctccattgtt acgagatccc cctagcagct ccttatgggg ttggctttgc taagaatggg
     5461 cctacccgct ggcaatacaa acggacaatc aatcaagtcg tccacagatg gggatcccac
     5521 acagtcccct ttctattaga accggataac atcaacggca aaacctgcac agcatcgcac
     5581 ctatgtcata atactcgatg ccacaatccc ttgcacttgt gctgggagtc actagacgac
     5641 aacaaaggca gaaactggtg cccgggtccc aacgggggat gtgtccatgc ggtggtttgt
     5701 ttaaggcagg gtccgttgta cggcccgggg gcgactgtgg caggtcctca acaaaggggc
     5761 agtcactttg tggtataagt cggcggccgc atcgatgccg tatacggtac ctttaagacc
     5821 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga
     5881 agggctaatt cactcccaaa gaagacaaga tctgcttttt gcctgtactg ggtctctctg
     5941 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     6001 tcaataaagc ttcagctgct cgagctagca gatctttttc cctctgccaa aaattatggg
     6061 gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt
     6121 gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat
     6181 catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg
     6241 gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct
     6301 gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt
     6361 tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga
     6421 tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatcc
     6481 ctcgacctgc agcccaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     6541 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     6601 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     6661 gggaaacctg tcgtgccagc ggatccgcat ctcaattagt cagcaaccat agtcccgccc
     6721 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     6781 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     6841 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctaac ttgtttattg
     6901 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     6961 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     7021 tccgctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     7081 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     7141 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     7201 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     7261 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     7321 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     7381 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     7441 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     7501 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     7561 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     7621 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     7681 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     7741 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     7801 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     7861 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     7921 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     7981 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     8041 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     8101 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     8161 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     8221 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     8281 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     8341 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     8401 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     8461 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     8521 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     8581 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     8641 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     8701 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     8761 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     8821 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     8881 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     8941 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     9001 agtgccacct gggtc
//
