LOCUS       40924_40257        4912 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSI-DST2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4912)
  AUTHORS   Kriz A, Schmid K, Baumgartner N, Ziegler U, Berger I, Ballmer-Hofer 
            K, Berger P.
  TITLE     A plasmid-based multigene expression system for mammalian cells
  JOURNAL   Nat Commun 1, 120 (2010)
  PUBMED    21081918
REFERENCE   2  (bases 1 to 4912)
  AUTHORS   Kriz A, Schmid K, Baumgartner N, Ziegler U, Berger I, Ballmer-Hofer 
            K, Berger P.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-OCT-2010) Laboratory of Biomolecular Research, Paul 
            Scherrer Institut, OFLC / 101, Villigen PSI CH-5232, Switzerland
REFERENCE   3  (bases 1 to 4912)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4912)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Commun 
            1, 120 (2010)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (11-OCT-2010) Laboratory of Biomolecular Research, Paul Scherrer 
            Institut, OFLC / 101, Villigen PSI CH-5232, Switzerland"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4912
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        71..374
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        375..578
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        623..641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             703..1410
                     /codon_start=1
                     /label=mTFP1
                     /note="monomeric teal (cyan) fluorescent protein (Ai et
                     al., 2006)"
                     /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
                     TNTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
                     EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLK
                     GDVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESA
                     VARNSTDGMDELYK"
     protein_bind    1469..1490
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        1505..1535
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1543..1559
                     /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)."
     primer_bind     1567..1583
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             1610..1632
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1757..2464
                     /codon_start=1
                     /label=GFP
                     /note="Aequorea victoria green fluorescent protein"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLCYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELY"
     polyA_signal    2657..2881
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             3357..4145
                     /codon_start=1
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      complement(4451..4832)
                     /direction=LEFT
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     protein_bind    complement(4874..4907)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 ttggcgcgcc tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
       61 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      121 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      181 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      241 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      301 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      361 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      421 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      481 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      541 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      601 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      661 gtttaaactt aagcttggta ccgagctcaa aggagatata ccatggtgag caagggcgag
      721 gagaccacaa tgggcgtaat caagcccgac atgaagatca agctgaagat ggagggcaac
      781 gtgaatggcc acgccttcgt gatcgagggc gagggcgagg gcaagcccta cgacggcacc
      841 aacaccatca acctggaggt gaaggaggga gcccccctgc ccttctccta cgacattctg
      901 accaccgcgt tcgcctacgg caacagggcc ttcaccaagt accccgacga catccccaac
      961 tacttcaagc agtccttccc cgagggctac tcttgggagc gcaccatgac cttcgaggac
     1021 aagggcatcg tgaaggtgaa gtccgacatc tccatggagg aggactcctt catctacgag
     1081 atacacctca agggcgagaa cttccccccc aacggccccg tgatgcagaa gaagaccacc
     1141 ggctgggacg cctccaccga gaggatgtac gtgcgcgacg gcgtgctgaa gggcgacgtc
     1201 aagcacaagc tgctgctgga gggcggcggc caccaccgcg ttgacttcaa gaccatctac
     1261 agggccaaga aggcggtgaa gctgcccgac tatcactttg tggaccaccg catcgagatc
     1321 ctgaaccacg acaaggacta caacaaggtg accgtttacg agagcgccgt ggcccgcaac
     1381 tccaccgacg gcatggacga gctgtacaag ggatcaggat caggatcaat gggaagagct
     1441 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     1501 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     1561 ttcacacagg aaacagctat gaccatgatt acgaatttct agaaataatt ttgtttaact
     1621 ttaagaagga gatatacata tggctagaat ccacagcaga tacaaagaac ttcttgccaa
     1681 aagagcagag cttcaaagaa aagtagagga acttcagaga gaaatttcca accgctcaac
     1741 ctcctcttca gagtctagca aaggagaaga actcttcact ggagttgtcc caattcttgt
     1801 tgaattagat ggtgatgtta acggccacaa gttctctgtc agtggagagg gtgaaggtga
     1861 tgcaacatac ggaaaactta ccctgaagtt catctgcact actggcaaac tgcctgttcc
     1921 atggccaaca ctagtcacta ctctgtgcta tggtgttcaa tgcttttcaa gatacccgga
     1981 tcatatgaaa cggcatgact ttttcaagag tgccatgccc gaaggttatg tacaggaaag
     2041 gaccatcttc ttcaaagatg acggcaacta caagacacgt gctgaagtca agtttgaagg
     2101 tgataccctt gttaatagaa tcgagttaaa aggtattgac ttcaaggaag atggcaacat
     2161 tctgggacac aaattggaat acaactataa ctcacacaat gtatacatca tggcagacaa
     2221 acaaaagaat ggaatcaaag tgaacttcaa gacccgccac aacattgaag atggaagcgt
     2281 tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg tccttttacc
     2341 agacaaccat tacctgtcca cacaatctgc cctttcgaaa gatcccaacg aaaagagaga
     2401 ccacatggtc cttcttgagt ttgtaacagc tgctgggatt acacatggca tggatgaact
     2461 gtacaactga gtaaattaca ttttatttac aatcactcga cgaagacttg atcacccggg
     2521 atctcgagcc atggtgctag cagctgatgc atagcatgcg gtaccgggag atgggggagg
     2581 ctaactgaaa cacggaaggg ctcttcctga ccgctcgagt ctagagggcc cgtttaaacc
     2641 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     2701 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     2761 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     2821 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     2881 gcttctgagg cggaaagaac cagctggggc tctaggttaa ttaagatcga tccctaggct
     2941 ttactaacta taacggtcct aaggtagcga ataccgtcga cctctagccc tgcaggaggt
     3001 ggcacttttc gggcagaata aatgatcata tcgtcaatta ttacctccac ggggagagcc
     3061 tgagcaaact ggcctcaggc atttgagaag cacacggtca cactgcttcc ggtagtcaat
     3121 aaaccggtaa gtagcgtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg
     3181 gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt ggggtacagt
     3241 ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt ttgatgttat
     3301 ggagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta aacatcatga
     3361 gggaagcggt gatcgccgaa gtatcgactc aactatcaga ggtagttggc gtcatcgagc
     3421 gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg gatggcggcc
     3481 tgaagccaca cagtgatatt gatttgctgg ttacggtgac cgtaaggctt gatgaaacaa
     3541 cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga gagagcgaga
     3601 ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg tggcgttatc
     3661 cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt gcaggtatct
     3721 tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca agagaacata
     3781 gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct gaacaggatc
     3841 tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac tgggctggcg
     3901 atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta accggcaaaa
     3961 tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc cagtatcagc
     4021 ccgtcatact tgaagctaga caggcttatc ttggacaaga agaagatcgc ttggcctcgc
     4081 gcgcagatca gttggaagaa tttgtccact acgtgaaagg cgagatcacc aaggtagtcg
     4141 gcaaataatg tctaacaatt cgttcaagcc gacggatatc gagctcgctt ggactcctag
     4201 gggttatgat agttattgct cagcggtggc agcagccaac tcagcttcct ttcgggcttt
     4261 gttagcagcc ggatcttcta ggctcaagca gtgatcagat ccagacatga taagatacat
     4321 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     4381 ttgtgtccct taacgtgagt ttaaagaaca tgtgagcacc tgagtgcttg cggcagcgtg
     4441 aagctaattc tgtcagccgt taagtgttcc tgtgtcactc aaaattgctt tgagaggctc
     4501 taagggcttc tcagtgcgtt acatccctgg cttgttgtcc acaaccgtta aaccttaaaa
     4561 gctttaaaag ccttatatat tctttttttt cttataaaac ttaaaacctt agaggctatt
     4621 taagttgctg atttatatta attttattgt tcaaacatga gagcttagta cgtgaaacat
     4681 gagagcttag tacgttagcc atgagagctt agtacgttag ccatgagggt ttagttcgtt
     4741 aaacatgaga gcttagtacg ttaaacatga gagcttagta cgtgaaacat gagagcttag
     4801 tacgtactat caacaggttg aactgctgat cgatcttttc tacggggcgg ccgcgtcagt
     4861 cagtcatcat accataactt cgtatagcat acattatacg aagttatctg gt
//
