LOCUS       40924_40262        4687 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSI-DST2cx, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4687)
  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 4687)
  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 4687)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4687)
  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..4687
                     /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             complement(2989..3777)
                     /codon_start=1
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      complement(4240..4628)
                     /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(4649..4682)
                     /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 tccctaggag
     2941 tccaagcgag ctcgatatcc gtcggcttga acgaattgtt agacattatt tgccgactac
     3001 cttggtgatc tcgcctttca cgtagtggac aaattcttcc aactgatctg cgcgcgaggc
     3061 caagcgatct tcttcttgtc caagataagc ctgtctagct tcaagtatga cgggctgata
     3121 ctgggccggc aggcgctcca ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc
     3181 ggttactgcg ctgtaccaaa tgcgggacaa cgtaagcact acatttcgct catcgccagc
     3241 ccagtcgggc ggcgagttcc atagcgttaa ggtttcattt agcgcctcaa atagatcctg
     3301 ttcaggaacc ggatcaaaga gttcctccgc cgctggacct accaaggcaa cgctatgttc
     3361 tcttgctttt gtcagcaaga tagccagatc aatgtcgatc gtggctggct cgaagatacc
     3421 tgcaagaatg tcattgcgct gccattctcc aaattgcagt tcgcgcttag ctggataacg
     3481 ccacggaatg atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga gaatctcgct
     3541 ctctccaggg gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc
     3601 atcaagcctt acggtcaccg taaccagcaa atcaatatca ctgtgtggct tcaggccgcc
     3661 atccactgcg gagccgtaca aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat
     3721 gacgccaact acctctgata gttgagtcga tacttcggcg atcaccgctt ccctcatgat
     3781 gtttaacttt gttttagggc gactgccctg ctgcgtaaca tcgttgctgc tccataacat
     3841 caaacatcga cccacggcgt aacgcgcttg ctgcttggat gcccgaggca tagactgtac
     3901 cccaaaaaaa cagtcataac aagccatgaa aaccgccact gcgccgttac caccgctgcg
     3961 ttcggtcaag gttctggacc agttgcgtga gcgcatacgc tacttaccgg tttattgact
     4021 accggaagca gtgtgaccgt gtgcttctca aatgcctgag gccagtttgc tcaggctctc
     4081 cccgtggagg taataattga cgatatgatc atttattctg cctcccagct gacattcatc
     4141 cggggtcagc accgtttctg cggactggct ttctacgtgt tccgcttcct ttagcagccc
     4201 ttgcgccctg agtgcttgcg gcagcgtgaa gctgaattct gtcagccgtt aagtgttcct
     4261 gtgtcactca aaattgcttt gagaggctct aagggcttct cagtgcgtta catccctggc
     4321 ttgttgtcca caaccgttaa accttaaaag ctttaaaagc cttatatatt cttttttttc
     4381 ttataaaact taaaacctta gaggctattt aagttgctga tttatattaa ttttattgtt
     4441 caaacatgag agcttagtac gtgaaacatg agagcttagt acgttagcca tgagagctta
     4501 gtacgttagc catgagggtt tagttcgtta aacatgagag cttagtacgt taaacatgag
     4561 agcttagtac gtgaaacatg agagcttagt acgtactatc aacaggttga actgctgatc
     4621 aacagatcct ctacgcggcc gcggtaccat aacttcgtat agcatacatt atacgaagtt
     4681 atctggt
//
