LOCUS       62056_6305        7632 bp DNA     circular SYN 15-MAY-2019
DEFINITION  Cloning vector pCHX50, complete sequence.
ACCESSION   MK448012
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7632)
  AUTHORS   Chassin H, Muller M, Tigges M, Scheller L, Lang M, Fussenegger M.
  TITLE     A modular degron library for synthetic circuits in mammalian cells
  JOURNAL   Nat Commun 10 (1), 2013 (2019)
  PUBMED    31043592
REFERENCE   2  (bases 1 to 7632)
  AUTHORS   Chassin H, Mueller M, Tigges M, Scheller L, Lang M, Fussenegger M.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-JAN-2019) D-BSSE, ETH Zurich, Mattenstrasse 26, Basel 
            4058, Switzerland
REFERENCE   3  (bases 1 to 7632)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun"; date: "2019"; volume: "10"; issue: "1"; pages: "2013"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (28-JAN-2019) D-BSSE, ETH Zurich, Mattenstrasse 26, Basel 4058, 
            Switzerland"
FEATURES             Location/Qualifiers
     source          1..7632
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        40..419
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        420..623
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        668..686
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    727..954
                     /label=UbVR
                     /note="UbVR"
     misc_feature    967..1194
                     /label=UbVR
                     /note="UbVR"
     misc_feature    1207..1434
                     /label=UbVR
                     /note="UbVR"
     CDS             1447..2448
                     /label=tTA
                     /note="tetracycline-controlled transactivator, comprising a
                     fusion of the tetracycline repressor TetR with the 
                     C-terminal activation domain of herpes simplex virus VP16"
     CDS             2461..3147
                     /label=Dendra2
                     /note="monomeric photoswitchable green-to-red fluorescent 
                     protein from Dendronephthya"
     misc_feature    3179..3191
                     /label=Activation Domain
                     /note="Activation Domain"
     misc_feature    3185..3191
                     /label=BB2
                     /note="BB2"
     polyA_signal    3232..3456
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3502..3930
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3944..4273
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4340..5131
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    5308..5441
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5478..5494)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5502..5518)
                     /label=lac operator
                     /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(5526..5556)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5571..5592)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5880..6465)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6639..7496)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7497..7601)
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctcca cgcgtggtac cctcgagttg acattgatta ttgagtagtt
       61 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
      121 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      181 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      241 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      301 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      361 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
      421 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
      481 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      541 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      601 gggaggtcta tataagcaga gctctctggc taactagaga acccactgct tactggctta
      661 tcgaaattaa tacgactcac tatagggaga cccaagctgt tcgaagcgga attcaccatg
      721 actagtcaga ttttcgtgaa gaccctgacc ggcaagacca tcaccctaga ggtggagccc
      781 agtgacacca tcgagaacgt gaaggccaag atccaggata aagagggcat cccccctgac
      841 cagcagaggc tgatctttgc cggcaagcag ctggaagatg gccgcaccct ctctgattac
      901 aacatccaga aggagtcaac cctgcacctg gtccttcgcc tgagaggtgt cagggctagt
      961 gctagtcaga ttttcgtgaa gaccctgacc ggcaagacca tcaccctaga ggtggagccc
     1021 agtgacacca tcgagaacgt gaaggccaag atccaggata aagagggcat cccccctgac
     1081 cagcagaggc tgatctttgc cggcaagcag ctggaagatg gccgcaccct ctctgattac
     1141 aacatccaga aggagtcaac cctgcacctg gtccttcgcc tgagaggtgt cagggctagt
     1201 gctagtcaga ttttcgtgaa gaccctgacc ggcaagacca tcaccctaga ggtggagccc
     1261 agtgacacca tcgagaacgt gaaggccaag atccaggata aagagggcat cccccctgac
     1321 cagcagaggc tgatctttgc cggcaagcag ctggaagatg gccgcaccct ctctgattac
     1381 aacatccaga aggagtcaac cctgcacctg gtccttcgcc tgagaggtgt cagggctagt
     1441 gctagttcca gattagataa aagtaaagtg attaacagcg cattagagct gcttaatgag
     1501 gtcggaatcg aaggtttaac aacccgtaaa ctcgcccaga agctaggtgt agagcagcct
     1561 acattgtatt ggcatgtaaa aaataagcgg gctttgctcg acgccttagc cattgagatg
     1621 ttagataggc accatactca cttttgccct ttagaagggg aaagctggca agatttttta
     1681 cgtaataacg ctaaaagttt tagatgtgct ttactaagtc atcgcgatgg agcaaaagta
     1741 catttaggta cacggcctac agaaaaacag tatgaaactc tcgaaaatca attagccttt
     1801 ttatgccaac aaggtttttc actagagaat gcattatatg cactcagcgc tgtggggcat
     1861 tttactttag gttgcgtatt ggaagatcaa gagcatcaag tcgctaaaga agaaagggaa
     1921 acacctacta ctgatagtat gccgccatta ttacgacaag ctatcgaatt atttgatcac
     1981 caaggtgcag agccagcctt cttattcggc cttgaattga tcatatgcgg attagaaaaa
     2041 caacttaaat gtgaaagtgg gtccgcgtac agccgcgcgc gtacgaaaaa caattacggg
     2101 tctaccatcg agggcctgct cgatctcccg gacgacgacg cccccgaaga ggcggggctg
     2161 gcggctccgc gcctgtcctt tctccccgcg ggacacacgc gcagactgtc gacggccccc
     2221 ccgaccgatg tcagcctggg ggacgagctc cacttagacg gcgaggacgt ggcgatggcg
     2281 catgccgacg cgctagacga tttcgatctg gacatgttgg gggacgggga ttccccgggt
     2341 ccgggattta ccccccacga ctccgccccc tacggcgctc tggatatggc cgacttcgag
     2401 tttgagcaga tgtttaccga tgcccttgga attgacgagt acggtggggc tagtgctagt
     2461 aacaccccgg gaattaacct gatcaaggag gacatgcgcg tgaaggtgca catggagggc
     2521 aacgtgaacg gccacgcctt cgtgatcgag ggcgagggca agggcaagcc ctacgagggc
     2581 acccagaccg ccaacctgac cgtgaaggag ggcgcccccc tgcccttcag ctacgacatc
     2641 ctgaccaccg ccgtgcacta cggcaaccgg gtgttcacca agtaccccga ggacatcccc
     2701 gactacttca agcagagctt ccccgagggc tacagctggg agcgcaccat gaccttcgag
     2761 gacaagggca tctgcaccat ccgcagcgac atcagcctgg agggcgactg cttcttccag
     2821 aacgtgcgct tcaagggcac caacttcccc cccaacggcc ccgtgatgca gaagaagacc
     2881 ctgaagtggg agcccagcac cgagaagctg cacgtgcgcg acggcctgct ggtgggcaac
     2941 atcaacatgg ccctgctgct ggagggcggc ggccactacc tgtgcgactt caagaccacc
     3001 tacaaggcca agaaggtggt gcagctgccc gacgcccact tcgtggacca ccgcatcgag
     3061 atcctgggca acgacagcga ctacaacaag gtgaagctgt acgagcacgc cgtggcccgc
     3121 tacagccccc tgcccagcca ggtgtggtcc ggcgacagcg gcgtgtacaa ggctagcgga
     3181 tccaccggtg tctagaaagc tttgaggccg gcctgcaggg atcagcctcg actgtgcctt
     3241 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     3301 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     3361 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     3421 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     3481 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3541 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     3601 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     3661 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     3721 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3781 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     3841 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     3901 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     3961 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4021 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4081 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     4141 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     4201 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     4261 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
     4321 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     4381 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     4441 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     4501 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4561 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4621 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     4681 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     4741 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     4801 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4861 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4921 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     4981 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     5041 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     5101 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     5161 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     5221 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     5281 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     5341 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     5401 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     5461 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     5521 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     5581 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5641 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     5701 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     5761 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5821 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5881 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5941 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     6001 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     6061 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     6121 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     6181 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     6241 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     6301 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     6361 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt
     6421 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6481 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6541 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6601 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6661 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6721 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6781 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6841 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6901 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6961 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     7021 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     7081 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     7141 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     7201 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     7261 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7321 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7381 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7441 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7501 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7561 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7621 ccacctgacg tc
//
