LOCUS       sequence-198277-        8390 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SNIPer plasmid, N terminal TEV fused to Frb.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8390)
  AUTHORS   Morgan CW, Diaz JE, Zeitlin SG, Gray DC, Wells JA
  TITLE     Engineered cellular gene-replacement platform for selective and 
            inducible proteolytic profiling.
  JOURNAL   Proc Natl Acad Sci U S A. 2015 Jul 7;112(27):8344-9. doi: 
            10.1073/pnas.1504141112. Epub 2015 Jun 23.
  PUBMED    26106156
REFERENCE   2  (bases 1 to 8390)
  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: 
            "10.1073/pnas.1504141112"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2015-07-7- 7"; volume: "112"; issue: "27"; pages: "8344-9"
COMMENT     Sequence Label: Frb-N-TEV
FEATURES             Location/Qualifiers
     source          1..8390
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        3..382
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        383..586
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             587..762
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    825..1024
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1225..1641
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1571..1593
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1609..1625
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     enhancer        1678..1981
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1982..2185
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2182..2206
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2332..2361
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2371..2640
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="EMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKETS
                     FNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISKQ"
     misc_feature    3131..3593
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     3414..3433
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3626..4225
                     /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(3626..3645)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4122..4142
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4463..5051
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4516..4536)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4934..4945)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5170..5595
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(5698..5720)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5856..5876)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5861..6190
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6041..6176
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6212..6231
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6312..6329)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6483..7071)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6563..6582)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7242..8033,8034..8102))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7865..7884
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8103..8207)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8275..8293
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
       61 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      121 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      181 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      241 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      301 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      361 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
      421 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
      481 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
      541 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcaat aaaagagccc
      601 acaacccctc actcggcgcg ccagtcttcc gatagactgc gtcgcccggg tacccgtatt
      661 cccaataaag cctcttgctg tttgcatccg aatcgtggtc tcgctgttcc ttgggagggt
      721 ctcctctgag tgattgacta cccacgacgg gggtctttca tttgggggct cgtccgggat
      781 ttggagaccc ctgcccaggg accaccgacc caccaccggg aggtaagctg gccagcaact
      841 tatctgtgtc tgtccgattg tctagtgtct atgtttgatg ttatgcgcct gcgtctgtac
      901 tagttagcta actagctctg tatctggcgg acccgtggtg gaactgacga gttctgaaca
      961 cccggccgca accctgggag acgtcccagg gactttgggg gccgtttttg tggcccgacc
     1021 tgaggaaggg agtcgatgtg gaatccgacc ccgtcaggat atgtggttct ggtaggagac
     1081 gagaacctaa aacagttccc gcctccgtct gaatttttgc tttcggtttg gaaccgaagc
     1141 cgcgcgtctt gtctgctgca gcgctgcagc atcgttctgt gttgtctctg tctgactgtg
     1201 tttctgtatt tgtctgaaaa ttagggccag actgttacca ctcccttaag tttgacctta
     1261 ggtcactgga aagatgtcga gcggatcgct cacaaccagt cggtagatgt caagaagaga
     1321 cgttgggtta ccttctgctc tgcagaatgg ccaaccttta acgtcggatg gccgcgagac
     1381 ggcaccttta accgagacct catcacccag gttaagatca aggtcttttc acctggcccg
     1441 catggacacc cagaccaggt cccctacatc gtgacctggg aagccttggc ttttgacccc
     1501 cctccctggg tcaagccctt tgtacaccct aagcctccgc ctcctcttcc tccatccgcc
     1561 ccgtctctcc cccttgaacc tcctcgttcg accccgcctc gatcctccct ttatccagcc
     1621 ctcactcctt ctctaggcgc cggaattgaa gatctggggg atcgatcctc tagagtccgt
     1681 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     1741 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     1801 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     1861 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     1921 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     1981 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     2041 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     2101 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     2161 gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcgcct ggagacgcca
     2221 tccacgctgt tttgacctcc atagaagaca ccgggaccga tccagcctcc gcggccggga
     2281 acggtgcatt ggaacggacc tgcaggaatt gatccgcggc cgcccaccat ggaacaaaag
     2341 ctgatctctg aggaggacct gctcgagatg gagatgtggc acgagggact cgaagaggcc
     2401 agcaggctgt actttggcga gaggaacgtc aagggcatgt tcgaagtgct ggagcccctc
     2461 catgcgatga tggaaagggg cccacagacc ctgaaggaga ccagcttcaa ccaggcttac
     2521 ggcagggacc tgatggaggc acaggaatgg tgcaggaagt acatgaagag cggcaacgtg
     2581 aaagacctga cccaggcgtg ggacctctac taccacgtgt tcaggaggat cagcaagcag
     2641 ggaggtggcg gaagcggcgg tgggggaagc ggcgagagcc ttttcaaggg cccgagggac
     2701 tacaacccga tctccagcac catctgtcac ctgaccaacg agagcgacgg tcacaccact
     2761 agtctgtacg gcatcggctt cggccccttc atcatcacca acaagcatct gttcaggagg
     2821 aataacggca cactgctggt gcaaagcctg cacggcgtgt tcaaagtgaa gaacacaacc
     2881 accctgcaac agcacctgat cgacggcagg gacatgatta tcatcaggat gcccaaggac
     2941 ttccccccct ttccccagaa actgaagttc agggagccac aaagggagga gcgaatctgc
     3001 ctggtgacca ccaacttcca gacctgagaa tccccccccc taacgttact ggccgaagcc
     3061 gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt
     3121 ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc
     3181 tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc
     3241 tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc
     3301 cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg
     3361 cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct
     3421 cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat
     3481 ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc
     3541 taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga taagcttgcc
     3601 acaacccaca aggagacgac cttccatgac cgagtacaag cccacggtgc gcctcgccac
     3661 ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     3721 cacgcgccac accgtcgacc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     3781 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     3841 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     3901 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     3961 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     4021 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     4081 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     4141 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     4201 catgacccgc aagcccggtg cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag
     4261 gagcgcacga ccccatggct ccgaccgaag ccacccgggg cggccccgcc gaccccgcac
     4321 ccgcccccga ggcccaccga ctctagtcga gggctgcagc gctgcagagg ccgagtgcag
     4381 aactgctcca aagggacctc aaggctttcc gagggacact aggctgactc catcgagcca
     4441 gtgtagagat aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     4501 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     4561 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     4621 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     4681 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     4741 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     4801 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     4861 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     4921 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     4981 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     5041 cgcctccccg catcggatat cagtggtcca ggctctagtt ttgactcaac aatatcacca
     5101 gctgaagcct atagagtacg agccatagat aaaataaaag attttattta gtctccagaa
     5161 aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt
     5221 ttgcaaggca tggaaaaata cataactgag aatagagaag ttcagatcaa ggtcaggaac
     5281 agatggaaca gggtcgaccc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
     5341 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
     5401 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggg gcgccagtcc
     5461 tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
     5521 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
     5581 cgggggtctt tcatttgggg gctcgtccgg gatcgggaga cccctgccca gggaccaccg
     5641 acccaccacc gggaggtaag ctggctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
     5701 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     5761 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca
     5821 gtcacgtagc gatagcggag tgtagatccg gctgtggaat gtgtgtcagt tagggtgtgg
     5881 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     5941 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     6001 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     6061 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     6121 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     6181 cttttgcaaa aagcttactg gcttaactat gcggcatcag agcagattgt actgagagtg
     6241 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     6301 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     6361 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     6421 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     6481 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     6541 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     6601 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     6661 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     6721 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     6781 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6841 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6901 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     6961 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     7021 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     7081 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     7141 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     7201 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     7261 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     7321 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     7381 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     7441 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     7501 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca
     7561 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     7621 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     7681 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     7741 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     7801 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca
     7861 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     7921 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     7981 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     8041 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     8101 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     8161 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     8221 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     8281 cgtatcacga ggccctttcg tcttcaagaa ttagcttggc cattgcatac gttgtatcca
     8341 tatcataata tgtacattta tattggctca tgtccaacat taccgccatg
//
