LOCUS       40924_40252        4308 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSI-DGB2x, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4308)
  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 4308)
  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 4308)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4308)
  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..4308
                     /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..1419
                     /codon_start=1
                     /label=EBFP2
                     /note="enhanced blue variant of GFP (Ai et al., 2007)"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     protein_bind    1478..1499
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        1514..1544
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1552..1568
                     /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     1576..1592
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             1619..1641
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1766..2473
                     /codon_start=1
                     /label=GFP
                     /note="Aequorea victoria green fluorescent protein"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLCYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELY"
     polyA_signal    2666..2890
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(3116..3646)
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      complement(3844..4225)
                     /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(4267..4300)
                     /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 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
      781 aagttcagcg tgaggggcga gggcgagggc gatgccacca acggcaagct gaccctgaag
      841 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgagc
      901 cacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
      961 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcacc
     1021 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tagtgaaccg catcgagctg
     1081 aagggcgtcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaacttc
     1141 aacagccaca acatctatat catggccgtc aagcagaaga acggcatcaa ggtgaacttc
     1201 aagatccgcc acaacgtgga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     1261 acccccatcg gcgacggccc cgtgctgctg cccgacagcc actacctgag cacccagtcc
     1321 gtgctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttccgcacc
     1381 gccgccggga tcactctcgg catggacgag ctgtacaagg gatcaggatc aggatcaatg
     1441 ggaagagctg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     1501 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     1561 gataacaatt tcacacagga aacagctatg accatgatta cgaatttcta gaaataattt
     1621 tgtttaactt taagaaggag atatacatat ggctagaatc cacagcagat acaaagaact
     1681 tcttgccaaa agagcagagc ttcaaagaaa agtagaggaa cttcagagag aaatttccaa
     1741 ccgctcaacc tcctcttcag agtctagcaa aggagaagaa ctcttcactg gagttgtccc
     1801 aattcttgtt gaattagatg gtgatgttaa cggccacaag ttctctgtca gtggagaggg
     1861 tgaaggtgat gcaacatacg gaaaacttac cctgaagttc atctgcacta ctggcaaact
     1921 gcctgttcca tggccaacac tagtcactac tctgtgctat ggtgttcaat gcttttcaag
     1981 atacccggat catatgaaac ggcatgactt tttcaagagt gccatgcccg aaggttatgt
     2041 acaggaaagg accatcttct tcaaagatga cggcaactac aagacacgtg ctgaagtcaa
     2101 gtttgaaggt gatacccttg ttaatagaat cgagttaaaa ggtattgact tcaaggaaga
     2161 tggcaacatt ctgggacaca aattggaata caactataac tcacacaatg tatacatcat
     2221 ggcagacaaa caaaagaatg gaatcaaagt gaacttcaag acccgccaca acattgaaga
     2281 tggaagcgtt caactagcag accattatca acaaaatact ccaattggcg atggccctgt
     2341 ccttttacca gacaaccatt acctgtccac acaatctgcc ctttcgaaag atcccaacga
     2401 aaagagagac cacatggtcc ttcttgagtt tgtaacagct gctgggatta cacatggcat
     2461 ggatgaactg tacaactgag taaattacat tttatttaca atcactcgac gaagacttga
     2521 tcacccggga tctcgagcca tggtgctagc agctgatgca tagcatgcgg taccgggaga
     2581 tgggggaggc taactgaaac acggaagggc tcttcctgac cgctcgagtc tagagggccc
     2641 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     2701 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     2761 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     2821 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     2881 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctaggttaat taagatcgat
     2941 ccctaggctt tactaactat aacggtcggt agcgaatacc gtcgacctct agccctgcag
     3001 gaggtggcac ttttcggtgg ccaagggcat ggtaaagact atattcgcgg cgttgtgaca
     3061 atttaccgaa caactccgcg gccgggaagc cgatctcggc ttgaacgaat tgttaggtgg
     3121 cggtacttgg gtcgatatca aagtgcatca cttcttcccg tatgcccaac tttgtataga
     3181 gagccactgc gggatcgtca ccgtaatctg cttgcacgta gatcacataa gcaccaagcg
     3241 cgttggcctc atgcttgagg agattgatga gcgcggtggc aatgccctgc ctccggtgct
     3301 cgccggagac tgcgagatca tagatataga tctcactacg cggctgctca aacttgggca
     3361 gaacgtaagc cgcgagagcg ccaacaaccg cttcttggtc gaaggcagca agcgcgatga
     3421 atgtcttact acggagcaag ttcccgaggt aatcggagtc cggctgatgt tgggagtagg
     3481 tggctacgtc tccgaactca cgaccgaaaa gatcaagagc agcccgcatg gatttgactt
     3541 ggtcagggcc gagcctacat gtgcgaatga tgcccatact tgagccacct aactttgttt
     3601 tagggcgact gccctgctgc gtaacatcgt tgctgctgcg taacatcgtt gctgctccat
     3661 aacatcaaac atcgacccac ggcgtaacgc gcttgctgct tggatgcccg aggcatagac
     3721 tgtacaaaaa aacagtcata acaagccatg aaaaccgcca ctgcgccgtt accaccgatc
     3781 ccttaacgtg agtttaaaga acatgtgagc acctgagtgc ttgcggcagc gtgaagctaa
     3841 ttctgtcagc cgttaagtgt tcctgtgtca ctcaaaattg ctttgagagg ctctaagggc
     3901 ttctcagtgc gttacatccc tggcttgttg tccacaaccg ttaaacctta aaagctttaa
     3961 aagccttata tattcttttt tttcttataa aacttaaaac cttagaggct atttaagttg
     4021 ctgatttata ttaattttat tgttcaaaca tgagagctta gtacgtgaaa catgagagct
     4081 tagtacgtta gccatgagag cttagtacgt tagccatgag ggtttagttc gttaaacatg
     4141 agagcttagt acgttaaaca tgagagctta gtacgtgaaa catgagagct tagtacgtac
     4201 tatcaacagg ttgaactgct gatcgatctt ttctacgggg cggccgcgtc agtcagtcat
     4261 cataccataa cttcgtatag catacattat acgaagttat ctggtttg
//
