LOCUS       62056_6285        7152 bp DNA     circular SYN 15-MAY-2019
DEFINITION  Cloning vector pCHX130, complete sequence.
ACCESSION   MK453495
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7152)
  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 7152)
  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 7152)
  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..7152
                     /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=UbS
                     /note="UbS"
     CDS             967..1968
                     /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             1981..2667
                     /label=Dendra2
                     /note="monomeric photoswitchable green-to-red fluorescent 
                     protein from Dendronephthya"
     misc_feature    2699..2711
                     /label=Activation Domain
                     /note="Activation Domain"
     misc_feature    2705..2711
                     /label=BB2
                     /note="BB2"
     polyA_signal    2752..2976
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3022..3450
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3464..3793
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3860..4651
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    4828..4961
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4998..5014)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5022..5038)
                     /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(5046..5076)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5091..5112)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5400..5985)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6159..7016)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7017..7121)
                     /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 cagtgctagt
      961 gctagttcca gattagataa aagtaaagtg attaacagcg cattagagct gcttaatgag
     1021 gtcggaatcg aaggtttaac aacccgtaaa ctcgcccaga agctaggtgt agagcagcct
     1081 acattgtatt ggcatgtaaa aaataagcgg gctttgctcg acgccttagc cattgagatg
     1141 ttagataggc accatactca cttttgccct ttagaagggg aaagctggca agatttttta
     1201 cgtaataacg ctaaaagttt tagatgtgct ttactaagtc atcgcgatgg agcaaaagta
     1261 catttaggta cacggcctac agaaaaacag tatgaaactc tcgaaaatca attagccttt
     1321 ttatgccaac aaggtttttc actagagaat gcattatatg cactcagcgc tgtggggcat
     1381 tttactttag gttgcgtatt ggaagatcaa gagcatcaag tcgctaaaga agaaagggaa
     1441 acacctacta ctgatagtat gccgccatta ttacgacaag ctatcgaatt atttgatcac
     1501 caaggtgcag agccagcctt cttattcggc cttgaattga tcatatgcgg attagaaaaa
     1561 caacttaaat gtgaaagtgg gtccgcgtac agccgcgcgc gtacgaaaaa caattacggg
     1621 tctaccatcg agggcctgct cgatctcccg gacgacgacg cccccgaaga ggcggggctg
     1681 gcggctccgc gcctgtcctt tctccccgcg ggacacacgc gcagactgtc gacggccccc
     1741 ccgaccgatg tcagcctggg ggacgagctc cacttagacg gcgaggacgt ggcgatggcg
     1801 catgccgacg cgctagacga tttcgatctg gacatgttgg gggacgggga ttccccgggt
     1861 ccgggattta ccccccacga ctccgccccc tacggcgctc tggatatggc cgacttcgag
     1921 tttgagcaga tgtttaccga tgcccttgga attgacgagt acggtggggc tagtgctagt
     1981 aacaccccgg gaattaacct gatcaaggag gacatgcgcg tgaaggtgca catggagggc
     2041 aacgtgaacg gccacgcctt cgtgatcgag ggcgagggca agggcaagcc ctacgagggc
     2101 acccagaccg ccaacctgac cgtgaaggag ggcgcccccc tgcccttcag ctacgacatc
     2161 ctgaccaccg ccgtgcacta cggcaaccgg gtgttcacca agtaccccga ggacatcccc
     2221 gactacttca agcagagctt ccccgagggc tacagctggg agcgcaccat gaccttcgag
     2281 gacaagggca tctgcaccat ccgcagcgac atcagcctgg agggcgactg cttcttccag
     2341 aacgtgcgct tcaagggcac caacttcccc cccaacggcc ccgtgatgca gaagaagacc
     2401 ctgaagtggg agcccagcac cgagaagctg cacgtgcgcg acggcctgct ggtgggcaac
     2461 atcaacatgg ccctgctgct ggagggcggc ggccactacc tgtgcgactt caagaccacc
     2521 tacaaggcca agaaggtggt gcagctgccc gacgcccact tcgtggacca ccgcatcgag
     2581 atcctgggca acgacagcga ctacaacaag gtgaagctgt acgagcacgc cgtggcccgc
     2641 tacagccccc tgcccagcca ggtgtggtcc ggcgacagcg gcgtgtacaa ggctagcgga
     2701 tccaccggtg tctagaaagc tttgaggccg gcctgcaggg atcagcctcg actgtgcctt
     2761 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     2821 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     2881 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     2941 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     3001 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3061 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     3121 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     3181 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     3241 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3301 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     3361 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     3421 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     3481 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3541 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3601 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     3661 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     3721 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     3781 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
     3841 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     3901 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     3961 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     4021 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4081 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4141 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     4201 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     4261 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     4321 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4381 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4441 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     4501 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     4561 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     4621 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     4681 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     4741 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     4801 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     4861 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     4921 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     4981 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     5041 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     5101 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5161 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     5221 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     5281 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5341 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5401 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5461 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5521 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5581 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5641 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5701 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5761 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5821 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5881 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt
     5941 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6001 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6061 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6121 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6181 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6241 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6301 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6361 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6421 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6481 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6541 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6601 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6661 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6721 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6781 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6841 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6901 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6961 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7021 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7081 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7141 ccacctgacg tc
//
