LOCUS       62056_13460        9948 bp DNA     circular SYN 15-FEB-2020
DEFINITION  Expression vector pKK172, complete sequence.
ACCESSION   MN811106
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9948)
  AUTHORS   Krawczyk K, Scheller L, Kim H, Fussenegger M.
  TITLE     Rewiring of endogenous signaling pathways to genomic targets for 
            therapeutic cell reprogramming
  JOURNAL   Nat Commun 11 (1), 608 (2020)
  PUBMED    32001704
REFERENCE   2  (bases 1 to 9948)
  AUTHORS   Krawczyk KK.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-DEC-2019) D-BSSE, ETH Zurich, Mattenstrasse 26, Basel 
            4058, Switzerland
REFERENCE   3  (bases 1 to 9948)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun"; date: "2020"; volume: "11"; issue: "1"; pages: "608"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (09-DEC-2019) D-BSSE, ETH Zurich, Mattenstrasse 26, Basel 4058, 
            Switzerland"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9948
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    complement(324..548)
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(662..1378)
                     /label=EGFP
                     /note="enhanced GFP"
     CDS             complement(2789..2815)
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
     CDS             complement(2822..2848)
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
     CDS             complement(2852..2878)
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
     misc_feature    complement(2894..2932)
                     /gene="GEAR NFAT"
                     /label=flexible linker
                     /note="flexible linker"
     misc_feature    complement(3924..4321)
                     /gene="GEAR NFAT"
                     /label=MCP
                     /note="MCP"
     promoter        complement(4353..5534)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     CDS             6167..6868
                     /label=dTomato
                     /note="dimeric variant of DsRed fluorescent protein (Shaner
                     et al., 2004)"
     CDS             6935..7330
                     /label=BSD
                     /note="blasticidin S deaminase"
     polyA_signal    7498..7553
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     rep_origin      complement(8116..8704)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8878..9735)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(9736..9840)
                     /label=AmpR promoter
ORIGIN
        1 ggatccctat acagttgaag tcggaagttt acatacactt aagttggagt cattaaaact
       61 cgtttttcaa ctactccaca aatttcttgt taacaaacaa tagttttggc aagtcagtta
      121 ggacatctac tttgtgcatg acacaagtca tttttccaac aattgtttac agacagatta
      181 tttcacttat aattcactgt atcacaattc cagtgggtca gaagtttaca tacactaagt
      241 tcgactcctc tgcagaatgc ggcgatgttt cggtaagggg tccgctatct agtaggccca
      301 gctggttctt tccgcctcag aagccataga gcccaccgca tccccagcat gcctgctatt
      361 gtcttcccaa tcctccccct tgctgtcctg ccccacccca ccccccagaa tagaatgaca
      421 cctactcaga caatgcgatg caatttcctc attttattag gaaaggacag tgggagtggc
      481 accttccagg gtcaaggaag gcacggggga ggggcaaaca acagatggct ggcaactaga
      541 aggcacagtc gaggctgatc agcgagctct agagaattga tccccaagct tggcctgaca
      601 ggccttaaac gggccctcta gactcgagcg gccgccactg tgctggatat ctgcagaatt
      661 ccttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga
      721 ccatgtgatc gcgcttctcg ttggggtctt tgctcagggc ggactgggtg ctcaggtagt
      781 ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg
      841 cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc ttgatgccgt
      901 tcttctgctt gtcggccatg atatagacgt tgtggctgtt gtagttgtac tccagcttgt
      961 gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca
     1021 gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga
     1081 agatggtgcg ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct
     1141 tcatgtggtc ggggtagcgg ctgaagcact gcacgccgta ggtcagggtg gtcacgaggg
     1201 tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt
     1261 aggtggcatc gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt
     1321 ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatgg
     1381 tggcgaccgg tggatcgatc tgaagcccca gaggcttgtt ctccatgtag ccgtggagct
     1441 gcaccacagg gtgtcctcct gttggggctt tgacagcgcc acggctgccc tccgtctcat
     1501 agtgggcccg gtgatggggc ttgggttgga cctcaatccg tagctcatag gagcccgact
     1561 gattggagag tggccactcg agtggtggga gggatgcagt cacagggatg ctgcagatgg
     1621 gaatggccgg caccaactgc ttgggccaag taggtggtac cagcaggatg gactctggag
     1681 cggaattcct cctctcctcc tgctcacatg gccccaggaa ctccacagta gggtagatgt
     1741 ggcgggccag gccagccttg gacggagcgg tggacacagg cgtcgggtca ggactggtct
     1801 tccatatctt ggaggggatc ccgcagggcg agtcggtggc cagggtgttg agggcatcca
     1861 tgagaacagc agagccggcc gtgggggggt acccagcggg gatgctgtcg tcctgcggtg
     1921 ccacgtgagg cgagggctgt ggcgaggggc tccgggagcg ctggggtgag gctgcgggca
     1981 gaggagcaac caaagcctct gcgcacgaat gcctccgctt ggcaccgggt gaggaggagc
     2041 gggatgccgg acggggcacg ggcgagtgtc ggcccaggca gctgtcctcg gcgaggctgg
     2101 ttcgaggtga cattattgga gaggttctgg gggaataatg agcagggatg ttttgaaact
     2161 ggggacacag gtcgtcgggc ccggcgttat tgggtgagac gcagggcgag gtgtaggggg
     2221 agaaggtgtc agaaatgaag ctggcagagg agccgctgct agcggggctc aagcaaagcg
     2281 gctcgcggta gccctcgtag ccgggcacgg gcagtgtgaa cctcgggctg gcggccacgc
     2341 cggccagggc cagggccggc tgctcggggg acaggccggc gtctctccca cggagggccc
     2401 cccctgcctg catcagttcg tgggatggag tgatctcgat ccgagggctc gggcccgaag
     2461 cccctgctgg cttgaccggg ctcagaaagt tctggaaccc tatactatcc ggctctccga
     2521 accggccagg gggctcgcca gagggactgg caagggggtt gcatggcttg aggccatagt
     2581 ccaggacatc atccgggtat gcgagtccgg agggtgagct gatggcctta tgtgcgatcg
     2641 gttcttcttc gatagggttc agatagtcat aatcgaagag gatggaaaag tccagctcgt
     2701 cttggggact gcccccgggc tcgtggccgg ggccgtcccc gccatcgggg tcgggctgcg
     2761 gctccgggct agagcggccg cactgagcag cgtaatctgg aacgtcatat ggataggatc
     2821 ctgcatagtc cgggacgtca tagggatagc ccgcatagtc aggaacatcg tatgggtaaa
     2881 agatcatggt ggagcctgat ccaccgctgc ctcctgaacc gccgcttccg cctgtacagg
     2941 agacagtggg gtccttggct ttgggaggct ccgagcctgt cagcagggag atggtggggt
     3001 cctcggcgaa gccgtcccct tcggagaagt aggagccctc tcccagctca aacagcaccg
     3061 gcaggtcgtt gctcccggtg tccacggagc cggggtccag caggaacagc ggctgcgctg
     3121 tgtagtgcac cagctgcttc cctgaatccg ggctgctgtt ctctgcctcg ggaggcctgg
     3181 ggggctcctg gggagacagg agctcttgga tactggccag gctgctgtca aggtcaggca
     3241 ggctcatgtc gggcacggtc accgaggggc tgaacaggtc cagcagggca ctggtgtcca
     3301 cgctgaagcc gcttccacct cctccctgcc cactagagga aatctgtgac agcagggcac
     3361 taaagtccat atcagcgatg cttgagaagt cttcatctcc ggacagccca ttaggcaggc
     3421 cgctggttcc caggggagtt ggagcggggt cggggggccg ctggctgccg gtcaccagcc
     3481 gggtaatggc ttcggggtac tccatcagca ttggttcggc tgtactatga gacatggaca
     3541 cgccctgatt cagcagctgc tgaaactcag agttgtccac ggaggccaga tctgtgaaca
     3601 ctccgggatc ggtgctgttc cccagcagag ctcccaggtc ctcatcagcg tcgaactgca
     3661 ggtgcagcag agcttcactc agagtcccct cgccggcctg tgtagacttg ggcactggag
     3721 cgctcagtga ctggggtggt cctggggtca gcacaggggc tggagcaggt ggctgggcca
     3781 gaggcaccat agcactagag ggcaccatag tctgggccag cactggagcg gagctagggg
     3841 ccagagccag ggcctggttg ctgatctgcc ctgaaggtcc gctacctcct cctccgcttc
     3901 ctcctcctcc gcttccacct cctccagcgc tgtagatacc tgagttagcg gcgatggcgg
     3961 aagggatagg attaccgtct ttgaggagcc cctgcattgc cttcacgatg agttcacagt
     4021 cagaattggt agcgaaaatt gggatagtga gctccatgtt caggtaggac ctccaagcgg
     4081 cgacaggcag ttcgactccg cccactgtct gggtagccac tttggggacc tccaccttga
     4141 tggtatactt tctcttctgg gcactagact gcctgacgct gcatgtcacc ttgtaggcct
     4201 ggctccgtga gttggagctg atccactctg ccaccccatt agcgaaatta gaaggagcca
     4261 ctgtcacatc ccctgtccca ccattgtcca cgagcacgaa ctgagtaaag tttgaagcca
     4321 tggtggcctc agaggccagc ttggggtagt tttcacgaca ccttaaatgg aagaaaaaaa
     4381 ctttgaacca ctgtctgagg cttgagaatg aaccaagatc caaactcaaa aagggcaaat
     4441 tccaaggaga attacatcaa gtgccaagct ggcctaactt cagtctccac ccactcagta
     4501 tggggaaact ccatcgcata aaacccctcc ccccaaccta aagacgacgt actccaaaag
     4561 ctcgagaact aatcgaggtg cctggacggc gcccggtact ccgtggagtc acatgaagcg
     4621 acggctgagg acggaaaggc ccttttcctt tgtgtgggtg actcacccgc ccgctctccc
     4681 gagcgccgcg tcctccattt tgagctccct gcagcagggc cgggaagcgg ccatctttcc
     4741 gctcacgcaa ctggtgccga ccgggccagc cttgccgccc agggcggggc gatacacggc
     4801 ggcgcgaggc caggcaccag agcaggccgg ccagcttgag actacccccg tccgattctc
     4861 ggtggccgcg ctcgcaggcc ccgcctcgcc gaacatgtgc gctgggacgc acgggccccg
     4921 tcgccgcccg cggccccaaa aaccgaaata ccagtgtgca gatcttggcc cgcatttaca
     4981 agactatctt gccagaaaaa aagcgtcgca gcaggtcatc aaaaatttta aatggctaga
     5041 gacttatcga aagcagcgag acaggcgcga aggtgccacc agattcgcac gcggcggccc
     5101 cagcgcccag gccaagcctc aactcaagca cgaggcgaag gggctcctta agcgcaaggc
     5161 ctcgaactct cccacccact tccaacccga agctcgggat caagaatcac gtactgcagc
     5221 caggggcgtg gaagtaattc aaggcacgca agggccataa cccgtaaaga ggccaggccc
     5281 gcgggaacca cacacggcac ttacctgtgt tctggcggca aacccgttgc gaaaaagaac
     5341 gttcacggcg actactgcac ttatatacgg ttctccccca ccctcgggaa aaaggcggag
     5401 ccagtacacg acatcacttt cccagtttac cccgcgccac cttctctagg caccggttca
     5461 attgccgacc cctcccccca acttctcggg gactgtgggc gatgtgcgct ctgcccactg
     5521 acgggcaccg gagcctctag gcgagaccct gtctcacaaa ataaagtaag cccggactga
     5581 gtgcggaaag gcgggcctgg cgggtctggt ctccccatgc gggccaccag aggccctgca
     5641 gccttcagtc gcttgaaggg gtaatggcgc ttccactcac aaacatggcg gacagagcgt
     5701 gtgaacgaga tgaacagccc ctcaaaaata tggccgccga ggctggacgg ccgtgcccca
     5761 gcagcaccgc ctccgcgccc cacgtgatct ctcgccgggc acagcgctga ccgcggaggt
     5821 ccaaccggaa gaatgtccgg attggacatt cggaagaggg cccgccttcc ctggggaatc
     5881 tctgcgcacg cgcagaacgc ttcgaccaat gaaaacacag gaagccgtcc gcgcaaccgc
     5941 gttgcgtcac ttctgccgcc cctgtttcaa ggtatatagc cgtagacgga acttcgcctt
     6001 tctctcggcc ttagcgccat ttttttgggt gagtgttttt tggttcctgc gttgggattc
     6061 cgtgtacaat ccatagacat ctgacctcgg cacttagcat catcacagca aactaactgt
     6121 agcctttctc tctttccctg tagaaacctc tgcacctgag gccaccatgt tgagcaaggg
     6181 cgaggaggtc atcaaagagt tcatgcgctt caaggtgcgc atggagggct ccatgaacgg
     6241 ccatgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
     6301 caagctgaag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccccca
     6361 gttcatgtac ggctccaagg cgtacgtgaa gcaccccgcc gacatccccg attacaagaa
     6421 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggtct
     6481 ggtgaccgtg acccaggact cctccctgca ggacggcacg ctgatctaca aggtgaagat
     6541 gcgcggcacc aacttccccc ccgacggccc cgtaatgcag aagaagacaa tgggctggga
     6601 ggcctccacc gagcgcctgt acccccgcga cggcgtgctg aagggcgaga tccaccaggc
     6661 cctgaagctg aaggacggcg gccactacct ggtggagttc aagaccatct acatggccaa
     6721 gaagcccgtg caactgcccg gctactacta cgtggacacc aagctggaca tcacctccca
     6781 caacgaggac tacaccatcg tggaacagta cgagcgctcc gagggccgcc accacctgtt
     6841 cctgtacggc atggacgagc tgtacaaggg cagtggagct actaacttca gcctgctgaa
     6901 gcaggctggt gacgtcgagg agaatcctgg ccccatggcc atgcctttgt ctcaagaaga
     6961 atccaccctc attgaaagag caacggctac aatcaacagc atccccatct ctgaagacta
     7021 cagcgtcgcc agcgcagctc tctctagcga cggccgcatc ttcactggtg tcaatgtata
     7081 tcattttact gggggacctt gtgcagaact cgtggtgctg ggcactgctg ctgctgcggc
     7141 agctggcaac ctgacttgta tcgtcgcgat cggaaatgag aacaggggca tcttgagccc
     7201 ctgcggacgg tgccgacagg tgcttctcga tctgcatcct gggatcaaag ccatagtgaa
     7261 ggacagtgat ggacagccga cggcagttgg gattcgtgaa ttgctgccct ctggttatgt
     7321 gtgggagggc taattcgaag gcctgtcgtg aattcactcc tcaggtgcag gctgcctatc
     7381 agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc
     7441 cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact tctggctaat
     7501 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
     7561 ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg
     7621 caacatatgc catatgctgg ctgccatgaa ctagctactc gggacccctt accgaaacat
     7681 cgccgcattc tgcagaggag tcgagtgtat gtaaacttct gacccactgg gaatgtgatg
     7741 aaagaaataa aagctgaaat gaatcattct ctctactatt attctgatat ttcacattct
     7801 taaaataaag tggtgatcct aactgaccta agacagggaa tttttactag gattaaatgt
     7861 caggaattgt gaaaaagtga gtttaaatgt atttggctaa ggtgtatgta aacttccgac
     7921 ttcaactgta tagggatccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     7981 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     8041 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     8101 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     8161 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     8221 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     8281 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     8341 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     8401 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     8461 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     8521 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     8581 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     8641 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     8701 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     8761 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     8821 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     8881 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     8941 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     9001 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     9061 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     9121 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     9181 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     9241 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     9301 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     9361 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     9421 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     9481 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     9541 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     9601 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     9661 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     9721 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     9781 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     9841 cacatttccc cgaaaagtgc cacctgatgc ggtgtgaaat accgcacaga tgcgtaagga
     9901 gaaaataccg catcaggaaa ttgtaagcgt taatattttg ttaaaatt
//
