LOCUS       40924_48138        5229 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pYL1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5229)
  AUTHORS   Liu Y, Charpin-El Hamri G, Ye H, Fussenegger M.
  TITLE     A synthetic free fatty acid-regulated transgene switch in mammalian 
            cells and mice
  JOURNAL   Nucleic Acids Res. (2018) In press
  PUBMED    30219861
REFERENCE   2  (bases 1 to 5229)
  AUTHORS   Liu Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-JUL-2018) Department of Biosystems Science and 
            Engineering, ETH Zurich, Mattenstrasse 26, basel 4058, Switzerland
REFERENCE   3  (bases 1 to 5229)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5229)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res. (2018) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (09-JUL-2018) Department of Biosystems Science and Engineering, ETH 
            Zurich, Mattenstrasse 26, basel 4058, Switzerland"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5229
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    14..21
                     /label=cAMP-response element (CRE)
                     /note="cAMP-response element (CRE)"
     misc_feature    40..44
                     /label=CRE
                     /note="CRE"
     misc_feature    50..54
                     /label=CRE
                     /note="CRE"
     misc_feature    72..79
                     /label=CRE
                     /note="CRE"
     misc_feature    109..118
                     /label=serum response factor binding element (SRE)
                     /note="serum response factor binding element (SRE)"
     misc_feature    132..141
                     /label=SRE
                     /note="SRE"
     misc_feature    155..164
                     /label=SRE
                     /note="SRE"
     protein_bind    189..218
                     /label=NFAT binding site
                     /note="NFAT binding site from the human interleukin 2
                     (IL-2) gene (Mattila et al., 1990)"
     protein_bind    219..248
                     /label=NFAT binding site
                     /bound_moiety="NFAT transcription factor"
                     /note="NFAT binding site from the human interleukin 2
                     (IL-2) gene (Mattila et al., 1990)"
     misc_feature    221..229
                     /label=NFAT
                     /note="NFAT"
     protein_bind    249..278
                     /label=NFAT binding site
                     /bound_moiety="NFAT transcription factor"
                     /note="NFAT binding site from the human interleukin 2
                     (IL-2) gene (Mattila et al., 1990)"
     misc_feature    251..259
                     /label=NFAT
                     /note="NFAT"
     promoter        291..322
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     CDS             344..1852
                     /codon_start=1
                     /label=SEAP
                     /note="secreted alkaline phosphatase from human placenta"
                     /translation="PTMLLLLLLLGLRLQLSLGIIPVEEENPDFWNREAAEALGAAKKL
                     QPAQTAAKNLIIFLGDGMGVSTVTAARILKGQKKDKLGPEIPLAMDRFPYVALSKTYNV
                     DKHVPDSGATATAYLCGVKGNFQTIGLSAAARFNQCNTTRGNEVISVMNRAKKAGKSVG
                     VVTTTRVQHASPAGTYAHTVNRNWYSDADVPASARQEGCQDIATQLISNMDIDVILGGG
                     RKYMFRMGTPDPEYPDDYSQGGTRLDGKNLVQEWLAKRQGARYVWNRTELMQASLDPSV
                     THLMGLFEPGDMKYEIHRDSTLDPSLMEMTEAALRLLSRNPRGFFLFVEGGRIDHGHHE
                     SRAYRALTETIMFDDAIERAGQLTSEEDTLSLVTADHSHVFSFGGYPLRGSSIFGLAPG
                     KARDRKAYTVLLYGNGPGYVLKDGARPDVTESESGSPEYRQQSAVPLDEETHAGEDVAV
                     FARGPQAHLVHGVQEQTFIAHVMAFAACLEPYTACDLAPPAGTTD"
     polyA_signal    complement(1910..2031)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2696..3284)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3458..4315)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4316..4420)
                     /label=AmpR promoter
     rep_origin      4447..4902
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     polyA_signal    5033..5081
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     misc_feature    5095..5186
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from
                     the human alpha-2 globin gene"
ORIGIN
        1 gcgcaccaga cagtgacgtc agctgccaga tcccatggcc gtcatactgt gacgtctttc
       61 agacacccca ttgacgtcaa tgggagaacc ccggtaccgg gaggatgtcc atattaggac
      121 atctaggatg tccatattag gacatctagg atgtccatat taggacatct cccggggagc
      181 tcgctagcgg aggaaaaact gtttcataca gaaggcgtgg aggaaaaact gtttcataca
      241 gaaggcgtgg aggaaaaact gtttcataca gaaggcgtag atctagactc tagagggtat
      301 ataatggaag ctcgaattcc agaagcttcg aatcgcgaat tcgcccacca tgctgctgct
      361 gctgctgctg ctgggcctga ggctacagct ctccctgggc atcatcccag ttgaggagga
      421 gaacccggac ttctggaacc gcgaggcagc cgaggccctg ggtgccgcca agaagctgca
      481 gcctgcacag acagccgcca agaacctcat catcttcctg ggcgatggga tgggggtgtc
      541 tacggtgaca gctgccagga tcctaaaagg gcagaagaag gacaaactgg ggcctgagat
      601 acccctggcc atggaccgct tcccatatgt ggctctgtcc aagacataca atgtagacaa
      661 acatgtgcca gacagtggag ccacagccac ggcctacctg tgcggggtca agggcaactt
      721 ccagaccatt ggcttgagtg cagccgcccg ctttaaccag tgcaacacga cacgcggcaa
      781 cgaggtcatc tccgtgatga atcgggccaa gaaagcaggg aagtcagtgg gagtggtaac
      841 caccacacga gtgcagcacg cctcgccagc cggcacctac gcccacacgg tgaaccgcaa
      901 ctggtactcg gacgccgacg tgcctgcctc ggcccgccag gaggggtgcc aggacatcgc
      961 tacgcagctc atctccaaca tggacattga cgtgatccta ggtggaggcc gaaagtacat
     1021 gtttcgcatg ggaaccccag accctgagta cccagatgac tacagccaag gtgggaccag
     1081 gctggacggg aagaatctgg tgcaggaatg gctggcgaag cgccagggtg cccggtatgt
     1141 gtggaaccgc actgagctca tgcaggcttc cctggacccg tctgtgaccc atctcatggg
     1201 tctctttgag cctggagaca tgaaatacga gatccaccga gactccacac tggacccctc
     1261 cctgatggag atgacagagg ctgccctgcg cctgctgagc aggaaccccc gcggcttctt
     1321 cctcttcgtg gagggtggtc gcatcgacca tggtcatcat gaaagcaggg cttaccgggc
     1381 actgactgag acgatcatgt tcgacgacgc cattgagagg gcgggccagc tcaccagcga
     1441 ggaggacacg ctgagcctcg tcactgccga ccactcccac gtcttctcct tcggaggcta
     1501 ccccctgcga gggagctcca tcttcgggct ggcccctggc aaggcccggg acaggaaggc
     1561 ctacacggtc ctcctatacg gaaacggtcc aggctatgtg ctcaaggacg gcgcccggcc
     1621 ggatgttacc gagagcgaga gcgggagccc cgagtatcgg cagcagtcag cagtgcccct
     1681 ggacgaagag acccacgcag gcgaggacgt ggcggtgttc gcgcgcggcc cgcaggcgca
     1741 cctggttcac ggcgtgcagg agcagacctt catagcgcac gtcatggcct tcgccgcctg
     1801 cctggagccc tacaccgcct gcgacctggc gccccccgcc ggcaccaccg acgccgcgca
     1861 cccgggttac tctagagtcg gggcggccgg ccgcttcgag cagacatgat aagatacatt
     1921 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     1981 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     2041 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag
     2101 taaaacctct acaaatgtgg taaaatcgat aaggatctga acgatggagc ggagaatggg
     2161 cggaactggg cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg
     2221 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
     2281 acctggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg
     2341 gggactttcc acaccctaac tgacacacat tccacagcgg atccgtcgac cgatgccctt
     2401 gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc
     2461 acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctcttccg
     2521 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     2581 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     2641 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2701 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2761 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2821 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2881 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2941 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3001 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3061 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3121 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3181 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3241 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3301 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3361 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     3421 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     3481 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     3541 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     3601 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     3661 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     3721 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     3781 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     3841 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     3901 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     3961 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     4021 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     4081 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     4141 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     4201 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     4261 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     4321 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     4381 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     4441 cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     4501 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     4561 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     4621 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     4681 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     4741 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     4801 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     4861 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttgccattcg ccattcaggc
     4921 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagcccaagc
     4981 taccatgata agtaagtaat attaaggtac gggaggtact tggagcggcc gcaataaaat
     5041 atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag tactaacata
     5101 cgctctccat caaaacaaaa cgaaacaaaa caaactagca aaataggctg tccccagtgc
     5161 aagtgcaggt gccagaacat ttctctatcg ataggtaccg agctcttacg cgtgctagcc
     5221 cgggctcga
//
