LOCUS       62056_6295        7146 bp DNA     circular SYN 15-MAY-2019
DEFINITION  Cloning vector pCHX181, complete sequence.
ACCESSION   MK453497
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7146)
  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 7146)
  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 7146)
  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..7146
                     /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=UbM
                     /note="UbM"
     CDS             961..1962
                     /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             1975..2661
                     /label=Dendra2
                     /note="monomeric photoswitchable green-to-red fluorescent 
                     protein from Dendronephthya"
     misc_feature    2693..2705
                     /label=Activation Domain
                     /note="Activation Domain"
     misc_feature    2699..2705
                     /label=BB2
                     /note="BB2"
     polyA_signal    2746..2970
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3016..3444
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3458..3787
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3854..4645
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    4822..4955
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4992..5008)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5016..5032)
                     /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(5040..5070)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5085..5106)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5394..5979)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6153..7010)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7011..7115)
                     /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 tgagaggtgg catggctagt
      961 tccagattag ataaaagtaa agtgattaac agcgcattag agctgcttaa tgaggtcgga
     1021 atcgaaggtt taacaacccg taaactcgcc cagaagctag gtgtagagca gcctacattg
     1081 tattggcatg taaaaaataa gcgggctttg ctcgacgcct tagccattga gatgttagat
     1141 aggcaccata ctcacttttg ccctttagaa ggggaaagct ggcaagattt tttacgtaat
     1201 aacgctaaaa gttttagatg tgctttacta agtcatcgcg atggagcaaa agtacattta
     1261 ggtacacggc ctacagaaaa acagtatgaa actctcgaaa atcaattagc ctttttatgc
     1321 caacaaggtt tttcactaga gaatgcatta tatgcactca gcgctgtggg gcattttact
     1381 ttaggttgcg tattggaaga tcaagagcat caagtcgcta aagaagaaag ggaaacacct
     1441 actactgata gtatgccgcc attattacga caagctatcg aattatttga tcaccaaggt
     1501 gcagagccag ccttcttatt cggccttgaa ttgatcatat gcggattaga aaaacaactt
     1561 aaatgtgaaa gtgggtccgc gtacagccgc gcgcgtacga aaaacaatta cgggtctacc
     1621 atcgagggcc tgctcgatct cccggacgac gacgcccccg aagaggcggg gctggcggct
     1681 ccgcgcctgt cctttctccc cgcgggacac acgcgcagac tgtcgacggc ccccccgacc
     1741 gatgtcagcc tgggggacga gctccactta gacggcgagg acgtggcgat ggcgcatgcc
     1801 gacgcgctag acgatttcga tctggacatg ttgggggacg gggattcccc gggtccggga
     1861 tttacccccc acgactccgc cccctacggc gctctggata tggccgactt cgagtttgag
     1921 cagatgttta ccgatgccct tggaattgac gagtacggtg gggctagtgc tagtaacacc
     1981 ccgggaatta acctgatcaa ggaggacatg cgcgtgaagg tgcacatgga gggcaacgtg
     2041 aacggccacg ccttcgtgat cgagggcgag ggcaagggca agccctacga gggcacccag
     2101 accgccaacc tgaccgtgaa ggagggcgcc cccctgccct tcagctacga catcctgacc
     2161 accgccgtgc actacggcaa ccgggtgttc accaagtacc ccgaggacat ccccgactac
     2221 ttcaagcaga gcttccccga gggctacagc tgggagcgca ccatgacctt cgaggacaag
     2281 ggcatctgca ccatccgcag cgacatcagc ctggagggcg actgcttctt ccagaacgtg
     2341 cgcttcaagg gcaccaactt cccccccaac ggccccgtga tgcagaagaa gaccctgaag
     2401 tgggagccca gcaccgagaa gctgcacgtg cgcgacggcc tgctggtggg caacatcaac
     2461 atggccctgc tgctggaggg cggcggccac tacctgtgcg acttcaagac cacctacaag
     2521 gccaagaagg tggtgcagct gcccgacgcc cacttcgtgg accaccgcat cgagatcctg
     2581 ggcaacgaca gcgactacaa caaggtgaag ctgtacgagc acgccgtggc ccgctacagc
     2641 cccctgccca gccaggtgtg gtccggcgac agcggcgtgt acaaggctag cggatccacc
     2701 ggtgtctaga aagctttgag gccggcctgc agggatcagc ctcgactgtg ccttctagtt
     2761 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     2821 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     2881 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     2941 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     3001 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3061 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     3121 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3181 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     3241 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3301 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     3361 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     3421 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     3481 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3541 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3601 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3661 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3721 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3781 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
     3841 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     3901 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     3961 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4021 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4081 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4141 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     4201 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4261 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     4321 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     4381 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     4441 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     4501 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     4561 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     4621 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     4681 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4741 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4801 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     4861 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4921 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     4981 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5041 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5101 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     5161 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5221 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5281 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5341 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5401 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5461 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5521 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5581 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5641 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5701 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5761 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5821 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5881 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tttttttgtt
     5941 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6001 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6061 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6121 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6181 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6241 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6301 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6361 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6421 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6481 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6541 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6601 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6661 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6721 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6781 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6841 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6901 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6961 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     7021 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7081 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     7141 gacgtc
//
