LOCUS       V016017                 7190 bp    DNA     circular SYN 15-FEB-2020
DEFINITION  Exported.
ACCESSION   V016017
VERSION     V016017
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7190)
  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 7190)
  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 7190)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "Nat
            Commun"; date: "2020"; volume: "11"; issue: "1"; pages: "608"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (09-DEC-2019) D-BSSE, ETH Zurich, Mattenstrasse 26, Basel 4058,
            Switzerland"
FEATURES             Location/Qualifiers
     source          1..7190
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             128..919
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    1096..1229
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1266..1282)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(1290..1306)
                     /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(1314..1344)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1359..1380)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1668..2253)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(2427..3284)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(3285..3389)
                     /label="AmpR promoter"
     rep_origin      3495..3630
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     misc_feature    3668..4024
                     /gene="GEAR SMAD2"
                     /label="MCP"
                     /note="MCP"
     CDS             4042..4755
                     /label="YPet"
                     /note="yellow fluorescent protein optimized for FRET with
                     CyPet (Nguyen and Daugherty, 2005)"
     CDS             4768..6165
                     /gene="SMAD2"
                     /label="Mothers against decapentaplegic homolog 2"
                     /note="Mothers against decapentaplegic homolog 2 from Homo
                     sapiens. Accession#: Q15796"
     polyA_signal    6210..6434
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6480..6908
                     /direction=RIGHT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6922..7190
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
       61 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
      121 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
      181 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
      241 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
      301 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
      361 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
      421 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
      481 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
      541 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
      601 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
      661 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
      721 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
      781 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
      841 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
      901 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
      961 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     1021 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     1081 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     1141 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     1201 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     1261 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     1321 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     1381 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     1441 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     1501 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     1561 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     1621 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     1681 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     1741 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     1801 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     1861 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     1921 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     1981 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2041 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2101 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2161 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt tgtttgcaag
     2221 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2281 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2341 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     2401 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     2461 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     2521 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     2581 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     2641 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     2701 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     2761 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     2821 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     2881 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     2941 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     3001 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     3061 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     3121 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     3181 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     3241 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     3301 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     3361 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     3421 gacggatcgg gagatctcca cgcgtggtac cctcgaggat ctgcgatctg catctcaatt
     3481 agtcagcaac catagtcccg cccctaactc cgcccagttc cgcccattct ccgccccatc
     3541 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     3601 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaattcga agcggaattc
     3661 accatgacta gtgcttctaa ctttactcag ttcgttctcg tcgacaatgg cggaactggc
     3721 gacgtgactg tcgccccaag caacttcgct aacgggatcg ctgaatggat cagctctaac
     3781 tcgcgttcac aggcttacaa agtaacctgt agcgttcgtc agagctctgc gcagaatcgc
     3841 aaatacacca tcaaagtcga ggtgcctaaa ggcgcctggc gttcgtactt aaatatggaa
     3901 ctaaccattc caattttcgc cacgaattcc gactgcgagc ttattgttaa ggcaatgcaa
     3961 ggtctcctaa aagatggaaa cccgattccc tcagcaatcg cagcaaactc cggcatctac
     4021 gcggctagtg gtggttctgg tgtgagcaaa ggcgaagagc tgttcaccgg cgtggtgccc
     4081 atcctggtgg agctggacgg cgacgtgaac ggccacaagt tcagcgtgag cggcgagggc
     4141 gagggcgacg ccacctacgg caagctgacc ctgaagctgc tgtgcaccac cggcaagctg
     4201 cccgtgccct ggcccaccct ggtgaccacc ctgggctacg gcgtgcagtg cttcgcccgg
     4261 taccccgacc acatgaagca gcacgacttc ttcaagagcg ccatgcccga gggctacgtg
     4321 caggagcgga ccatcttctt caaggacgac ggcaactaca agacccgggc cgaggtgaag
     4381 ttcgagggcg acaccctggt gaaccggatc gagctgaagg gcatcgactt caaggaggac
     4441 ggcaacatcc tgggccacaa gctggagtac aactacaaca gccacaacgt gtacatcacc
     4501 gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccggcacaa catcgaggac
     4561 ggcggcgtgc agctggccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     4621 ctgctgcccg acaaccacta cctgagctac cagagcgccc tgttcaagga ccccaacgag
     4681 aagcgggacc acatggtgct gctggagttc ctgaccgccg ccggcatcac cgagggcatg
     4741 gacgagctgt acaaggctag tggtggttcg tccatcttgc cattcacgcc gccagttgtg
     4801 aagagactgc tgggatggaa gaagtcagct ggtgggtctg gaggagcagg cggaggagag
     4861 cagaatgggc aggaagaaaa gtggtgtgag aaagcagtga aaagtctggt gaagaagcta
     4921 aagaaaacag gacgattaga tgagcttgag aaagccatca ccactcaaaa ctgtaatact
     4981 aaatgtgtta ccataccaag cacttgctct gaaatttggg gactgagtac accaaatacg
     5041 atagatcagt gggatacaac aggcctttac agcttctctg aacaaaccag gtctcttgat
     5101 ggtcgtctcc aggtatccca tcgaaaagga ttgccacatg ttatatattg ccgattatgg
     5161 cgctggcctg atcttcacag tcatcatgaa ctcaaggcaa ttgaaaactg cgaatatgct
     5221 tttaatctta aaaaggatga agtatgtgta aacccttacc actatcagag agttgagaca
     5281 ccagttttgc ctccagtatt agtgccccga cacaccgaga tcctaacaga acttccgcct
     5341 ctggatgact atactcactc cattccagaa aacactaact tcccagcagg aattgagcca
     5401 cagagtaatt atattccaga aacgccacct cctggatata tcagtgaaga tggagaaaca
     5461 agtgaccaac agttgaatca aagtatggac acaggctctc cagcagaact atctcctact
     5521 actctttccc ctgttaatca tagcttggat ttacagccag ttacttactc agaacctgca
     5581 ttttggtgtt cgatagcata ttatgaatta aatcagaggg ttggagaaac cttccatgca
     5641 tcacagccct cactcactgt agatggcttt acagacccat caaattcaga gaggttctgc
     5701 ttaggtttac tctccaatgt taaccgaaat gccacggtag aaatgacaag aaggcatata
     5761 ggaagaggag tgcgcttata ctacataggt ggggaagttt ttgctgagtg cctaagtgat
     5821 agtgcaatct ttgtgcagag ccccaattgt aatcagagat atggctggca ccctgcaaca
     5881 gtgtgtaaaa ttccaccagg ctgtaatctg aagatcttca acaaccagga atttgctgct
     5941 cttctggctc agtctgttaa tcagggtttt gaagccgtct atcagctaac tagaatgtgc
     6001 accataagaa tgagttttgt gaaagggtgg ggagcagaat accgaaggca gacggtaaca
     6061 agtactcctt gctggattga acttcatctg aatggacctc tacagtggtt ggacaaagta
     6121 ttaactcaga tgggatcccc ttcagtgcgt tgctcaagca tgtcatagtc tagaaagctt
     6181 tgaggccggc ctgcagggat cagcctcgac tgtgccttct agttgccagc catctgttgt
     6241 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     6301 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     6361 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     6421 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     6481 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     6541 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     6601 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     6661 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     6721 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     6781 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     6841 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     6901 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     6961 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     7021 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     7081 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     7141 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
//
