LOCUS       40924_38398        5594 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Mammalian expression vector pSA89, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5594)
  AUTHORS   Auslaender S, Auslaender D, Mueller M, Wieland M, Fussenegger M.
  TITLE     Programmable single-cell mammalian biocomputers
  JOURNAL   Nature 487 (7405), 123-127 (2012)
  PUBMED    22722847
REFERENCE   2  (bases 1 to 5594)
  AUTHORS   Auslaender S, Auslaender D, Mueller M, Wieland M, Fussenegger M.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-FEB-2012) Department of Biosystems Science and 
            Engineering, D-BSSE, ETH Zurich, Mattenstrasse 26, Basel, BS 4058, 
            Switzerland
REFERENCE   3  (bases 1 to 5594)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5594)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature"; 
            date: "2012"; volume: "487"; issue: "7405"; pages: "123-127"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-FEB-2012) Department of Biosystems Science and Engineering, 
            D-BSSE, ETH Zurich, Mattenstrasse 26, Basel, BS 4058, Switzerland"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5594
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        188..226
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        348..366
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             422..1264
                     /codon_start=1
                     /label=d2EYFP
                     /note="EYFP destabilized by residues 422-461 of mouse
                     ornithine decarboxylase, giving an in vivo half-life of ~2 
                     hours"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKKLSHGFPPAVAAQDDGTLPMSCAQESGMDRHPAACASARINV
                     "
     polyA_signal    1316..1540
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1586..2014
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2028..2357
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2424..3215
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    3392..3525
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3562..3578)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3586..3602)
                     /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(3610..3640)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3655..3676)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3964..4549)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4723..5580)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(join(5581..5594,1..91))
                     /label=AmpR promoter
ORIGIN
        1 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
       61 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
      121 tcgattgaat ataaccgacg tgactgttac atttaggcct gcaggtcgag ctcggtaccc
      181 gggtcgaggt aggcgtgtac ggtgggaggc ctatataagc agagctcgtt tagtgaaccg
      241 tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagac accgggaccg
      301 atccagcctc cgcggaattc acccactgct tactggctta tcgaaattaa tacgactcac
      361 tatagggaga cccacctaga tctccgtgag gatcacccac ggggatccac cggtcgccac
      421 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
      481 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
      541 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
      601 cctcgtgacc accttcggct acggcctgca gtgcttcgcc cgctaccccg accacatgaa
      661 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
      721 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
      781 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
      841 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
      901 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
      961 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     1021 ctacctgagc taccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     1081 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagaa
     1141 gcttagccat ggcttcccgc cggcggtggc ggcgcaggat gatggcacgc tgcccatgtc
     1201 ttgtgcccag gagagcggga tggaccgtca ccctgcagcc tgtgcttctg ctaggatcaa
     1261 tgtgtaggcg gccgcgactc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg
     1321 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     1381 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     1441 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     1501 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     1561 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     1621 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     1681 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     1741 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     1801 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     1861 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     1921 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     1981 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     2041 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     2101 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2161 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     2221 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     2281 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     2341 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     2401 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     2461 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     2521 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     2581 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     2641 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     2701 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     2761 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     2821 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     2881 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     2941 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     3001 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     3061 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     3121 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     3181 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     3241 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     3301 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     3361 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     3421 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     3481 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     3541 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     3601 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     3661 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     3721 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     3781 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3841 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3901 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3961 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4021 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4081 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4141 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4201 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4261 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4321 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4381 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4441 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4501 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4561 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4621 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4681 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4741 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4801 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4861 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4921 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4981 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5041 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5101 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5161 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5221 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5281 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5341 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5401 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5461 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5521 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5581 actcttcctt tttc
//
