LOCUS       40924_40227        4625 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSI-DAL2x, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4625)
  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 4625)
  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 4625)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4625)
  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..4625
                     /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=mEYFP
                     /note="enhanced YFP with monomerizing A206K mutation
                     (Zacharias et al., 2002)"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     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"
     promoter        3027..3131
                     /label=AmpR promoter
     CDS             3132..3989
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      complement(4164..4545)
                     /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(4587..4620)
                     /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 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
      841 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccttcggc
      901 tacggcctga tgtgcttcgc ccgctacccc gaccacatga agcgtcacga cttcttcaag
      961 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     1021 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     1081 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     1141 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     1201 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     1261 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc
     1321 aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     1381 gccgccggga tcactcacgg 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 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3061 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3121 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3181 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3241 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3301 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     3361 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     3421 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     3481 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     3541 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     3601 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     3661 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     3721 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     3781 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     3841 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     3901 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     3961 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     4021 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4081 ataatctcat gaccaaaatc ccttaacgtg agtttaaaga acatgtgagc acctgagtgc
     4141 ttgcggcagc gtgaagctaa ttctgtcagc cgttaagtgt tcctgtgtca ctcaaaattg
     4201 ctttgagagg ctctaagggc ttctcagtgc gttacatccc tggcttgttg tccacaaccg
     4261 ttaaacctta aaagctttaa aagccttata tattcttttt tttcttataa aacttaaaac
     4321 cttagaggct atttaagttg ctgatttata ttaattttat tgttcaaaca tgagagctta
     4381 gtacgtgaaa catgagagct tagtacgtta gccatgagag cttagtacgt tagccatgag
     4441 ggtttagttc gttaaacatg agagcttagt acgttaaaca tgagagctta gtacgtgaaa
     4501 catgagagct tagtacgtac tatcaacagg ttgaactgct gatcgatctt ttctacgggg
     4561 cggccgcgtc agtcagtcat cataccataa cttcgtatag catacattat acgaagttat
     4621 ctggt
//
