LOCUS       62056_20115        5784 bp DNA     circular SYN 15-FEB-2020
DEFINITION  Expression vector pSP20, complete sequence.
ACCESSION   MN811117
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5784)
  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 5784)
  AUTHORS   Saxena P.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-DEC-2019) D-BSSE, ETH Zurich, Mattenstrasse 26, Basel 
            4058, Switzerland
REFERENCE   3  (bases 1 to 5784)
  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..5784
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    37..928
                     /label=human insulin promoter fragment
                     /note="human insulin promoter fragment"
     CDS             935..2443
                     /label=SEAP
                     /note="secreted alkaline phosphatase from human placenta"
     polyA_signal    complement(2501..2622)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3287..3875)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4049..4906)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4907..5011)
                     /label=AmpR promoter
     rep_origin      5038..5493
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     misc_feature    complement(5495..5563)
                     /label=LacZ alpha
                     /note="LacZ alpha"
     polyA_signal    5624..5672
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     misc_feature    5686..5777
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from
                     the human alpha-2 globin gene"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagtcc ctcactccca ctctcccacc
       61 cccaccacct tggcccatcc atggcggcat cttgggccat ccgggactgg ggacaggggt
      121 cctggggaca ggggtgtggg gacaggggtc ctggggacag gggtctgggg acaggggtcc
      181 tggggacagg ggtgtgggga caggggtgtg gggacagggg tgtggggaca ggggtcctgg
      241 ggacaggggt ctggggacag gggtctgagg acaggggtgt ggggacaggg gtgtggggac
      301 aggggtgtgg ggacaggggt gtggggacag gggtctgggg acaggggtcc gggggacagg
      361 ggtgtgggga caggggtgtg gggacagggg tgtggggaca ggggtctggg gacaggggtg
      421 tggggacagg ggtcctgggg acaggggtgt ggggataggg gtgtggggac aggggtgtgg
      481 ggacaggggt gtggggacag gggtctgggg acagcagcgc aaagagcccc gccctgcagc
      541 ctccagctct cctggtctaa tgtggaaagt ggcccaggtg agggctttgc tctcctggag
      601 acatttgccc ccagctgtga gcagggacag gtctggccac cgggcccctg gttaagactc
      661 taatgacccg ctggtcctga ggaagaggtg ctgacgacca aggagatctt cccacagacc
      721 cagcaccagg gaaatggtcc ggaaattgca gcctcagccc ccagccatct gccgaccccc
      781 ccaccccagg ccctaatggg ccaggcggca ggggttgaga ggtaggggag atgggctctg
      841 agactataaa gccagcgggg gcccagcagc cctcagccct ccaggacagg ctgcatcaga
      901 agaggccatc aagcaggtct gttccaagaa ttcgcccacc atgctgctgc tgctgctgct
      961 gctgggcctg aggctacagc tctccctggg catcatccca gttgaggagg agaacccgga
     1021 cttctggaac cgcgaggcag ccgaggccct gggtgccgcc aagaagctgc agcctgcaca
     1081 gacagccgcc aagaacctca tcatcttcct gggcgatggg atgggggtgt ctacggtgac
     1141 agctgccagg atcctaaaag ggcagaagaa ggacaaactg gggcctgaga tacccctggc
     1201 catggaccgc ttcccatatg tggctctgtc caagacatac aatgtagaca aacatgtgcc
     1261 agacagtgga gccacagcca cggcctacct gtgcggggtc aagggcaact tccagaccat
     1321 tggcttgagt gcagccgccc gctttaacca gtgcaacacg acacgcggca acgaggtcat
     1381 ctccgtgatg aatcgggcca agaaagcagg gaagtcagtg ggagtggtaa ccaccacacg
     1441 agtgcagcac gcctcgccag ccggcaccta cgcccacacg gtgaaccgca actggtactc
     1501 ggacgccgac gtgcctgcct cggcccgcca ggaggggtgc caggacatcg ctacgcagct
     1561 catctccaac atggacattg acgtgatcct aggtggaggc cgaaagtaca tgtttcgcat
     1621 gggaacccca gaccctgagt acccagatga ctacagccaa ggtgggacca ggctggacgg
     1681 gaagaatctg gtgcaggaat ggctggcgaa gcgccagggt gcccggtatg tgtggaaccg
     1741 cactgagctc atgcaggctt ccctggaccc gtctgtgacc catctcatgg gtctctttga
     1801 gcctggagac atgaaatacg agatccaccg agactccaca ctggacccct ccctgatgga
     1861 gatgacagag gctgccctgc gcctgctgag caggaacccc cgcggcttct tcctcttcgt
     1921 ggagggtggt cgcatcgacc atggtcatca tgaaagcagg gcttaccggg cactgactga
     1981 gacgatcatg ttcgacgacg ccattgagag ggcgggccag ctcaccagcg aggaggacac
     2041 gctgagcctc gtcactgccg accactccca cgtcttctcc ttcggaggct accccctgcg
     2101 agggagctcc atcttcgggc tggcccctgg caaggcccgg gacaggaagg cctacacggt
     2161 cctcctatac ggaaacggtc caggctatgt gctcaaggac ggcgcccggc cggatgttac
     2221 cgagagcgag agcgggagcc ccgagtatcg gcagcagtca gcagtgcccc tggacgaaga
     2281 gacccacgca ggcgaggacg tggcggtgtt cgcgcgcggc ccgcaggcgc acctggttca
     2341 cggcgtgcag gagcagacct tcatagcgca cgtcatggcc ttcgccgcct gcctggagcc
     2401 ctacaccgcc tgcgacctgg cgccccccgc cggcaccacc gacgccgcgc acccgggtta
     2461 ctctagagtc ggggcggccg gccgcttcga gcagacatga taagatacat tgatgagttt
     2521 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     2581 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     2641 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc
     2701 tacaaatgtg gtaaaatcga taaggatctg aacgatggag cggagaatgg gcggaactgg
     2761 gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga
     2821 gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg
     2881 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
     2941 cacaccctaa ctgacacaca ttccacagcg gatccgtcga ccgatgccct tgagagcctt
     3001 caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac
     3061 tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctcttcc gcttcctcgc
     3121 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     3181 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     3241 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3301 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3361 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3421 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3481 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3541 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3601 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3661 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3721 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3781 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3841 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3901 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3961 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4021 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     4081 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4141 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     4201 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     4261 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4321 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     4381 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     4441 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     4501 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4561 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4621 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4681 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     4741 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     4801 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     4861 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     4921 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4981 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     5041 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     5101 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     5161 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     5221 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     5281 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     5341 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     5401 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     5461 cgaattttaa caaaatatta acgtttacaa tttcccattc gccattcagg ctgcgcaact
     5521 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagcccaag ctaccatgat
     5581 aagtaagtaa tattaaggta cgggaggtac ttggagcggc cgcaataaaa tatctttatt
     5641 ttcattacat ctgtgtgttg gttttttgtg tgaatcgata gtactaacat acgctctcca
     5701 tcaaaacaaa acgaaacaaa acaaactagc aaaataggct gtccccagtg caagtgcagg
     5761 tgccagaaca tttctctatc gata
//
