LOCUS       40924_40242        4471 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSI-DCP2x, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4471)
  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 4471)
  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 4471)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4471)
  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..4471
                     /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..1380
                     /codon_start=1
                     /label=mPlum
                     /note="far-red monomeric derivative of DsRed fluorescent
                     protein (Wang et al., 2004)"
                     /translation="MVSKGEEVIKEFMRFKEHMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     RLKVTKGGPLPFAWDILSPQIMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGEFIYKVKVRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKGEMK
                     MRLRLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRH
                     STGA"
     protein_bind    1433..1454
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        1469..1499
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1507..1523
                     /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     1531..1547
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             1574..1596
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1721..2428
                     /codon_start=1
                     /label=GFP
                     /note="Aequorea victoria green fluorescent protein"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLCYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELY"
     polyA_signal    2621..2845
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        3020..3122
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             3123..3779
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     rep_origin      complement(4007..4388)
                     /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(4430..4463)
                     /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 gaggtcatca aggagttcat gcgcttcaag gagcacatgg agggctccgt gaacggccac
      781 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccagg
      841 ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagatc
      901 atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg
      961 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg
     1021 accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt gaaggtgcgc
     1081 ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc
     1141 tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatgaa gatgaggctg
     1201 aggctgaagg acggcggcca ctacgacgcc gaggtcaaga ccacctacat ggccaagaag
     1261 cccgtgcagc tgcccggcgc ctacaagacc gacatcaagc tggacatcac ctcccacaac
     1321 gaggactaca ccatcgtgga acagtacgag cgcgccgagg gccgccactc caccggcgcc
     1381 ggatcaggat caatgggaag agctggaaag cgggcagtga gcgcaacgca attaatgtga
     1441 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     1501 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgaat
     1561 ttctagaaat aattttgttt aactttaaga aggagatata catatggcta gaatccacag
     1621 cagatacaaa gaacttcttg ccaaaagagc agagcttcaa agaaaagtag aggaacttca
     1681 gagagaaatt tccaaccgct caacctcctc ttcagagtct agcaaaggag aagaactctt
     1741 cactggagtt gtcccaattc ttgttgaatt agatggtgat gttaacggcc acaagttctc
     1801 tgtcagtgga gagggtgaag gtgatgcaac atacggaaaa cttaccctga agttcatctg
     1861 cactactggc aaactgcctg ttccatggcc aacactagtc actactctgt gctatggtgt
     1921 tcaatgcttt tcaagatacc cggatcatat gaaacggcat gactttttca agagtgccat
     1981 gcccgaaggt tatgtacagg aaaggaccat cttcttcaaa gatgacggca actacaagac
     2041 acgtgctgaa gtcaagtttg aaggtgatac ccttgttaat agaatcgagt taaaaggtat
     2101 tgacttcaag gaagatggca acattctggg acacaaattg gaatacaact ataactcaca
     2161 caatgtatac atcatggcag acaaacaaaa gaatggaatc aaagtgaact tcaagacccg
     2221 ccacaacatt gaagatggaa gcgttcaact agcagaccat tatcaacaaa atactccaat
     2281 tggcgatggc cctgtccttt taccagacaa ccattacctg tccacacaat ctgccctttc
     2341 gaaagatccc aacgaaaaga gagaccacat ggtccttctt gagtttgtaa cagctgctgg
     2401 gattacacat ggcatggatg aactgtacaa ctgagtaaat tacattttat ttacaatcac
     2461 tcgacgaaga cttgatcacc cgggatctcg agccatggtg ctagcagctg atgcatagca
     2521 tgcggtaccg ggagatgggg gaggctaact gaaacacgga agggctcttc ctgaccgctc
     2581 gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag
     2641 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     2701 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     2761 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     2821 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     2881 ttaattaaga tcgatcccta ggctttacta actataacgg tcggtagcga ataccgtcga
     2941 cctctagccc tgcaggaggt ggcacttttc gggttgatac cgggaagccc tgggccaact
     3001 tttggcgaaa atgagacgtt gatcggcacg taagaggttc caactttcac cataatgaaa
     3061 taagatcact accgggcgta ttttttgagt tatcgagatt ttcaggagct aaggaagcta
     3121 aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag
     3181 aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg
     3241 atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta
     3301 ttcacattct tgcccgcctg atgaatgctc atccggaatt ccgtatggca atgaaagacg
     3361 gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg
     3421 aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat
     3481 attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg
     3541 agaatatgtt tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg
     3601 tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag
     3661 gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc
     3721 atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt
     3781 aattttttta aggcagttat tggtgccctt aaacgcctgg ttgctacgcc tgaataagtg
     3841 ataataagcg gatgaatggc agaaattcga aagcaaattc gacccggtcg tcggttcagg
     3901 gcagggtcgt taaatagccg cttatgtcta ttgctggttt atcccttaac gtgagtttaa
     3961 agaacatgtg agcacctgag tgcttgcggc agcgtgaagc taattctgtc agccgttaag
     4021 tgttcctgtg tcactcaaaa ttgctttgag aggctctaag ggcttctcag tgcgttacat
     4081 ccctggcttg ttgtccacaa ccgttaaacc ttaaaagctt taaaagcctt atatattctt
     4141 ttttttctta taaaacttaa aaccttagag gctatttaag ttgctgattt atattaattt
     4201 tattgttcaa acatgagagc ttagtacgtg aaacatgaga gcttagtacg ttagccatga
     4261 gagcttagta cgttagccat gagggtttag ttcgttaaac atgagagctt agtacgttaa
     4321 acatgagagc ttagtacgtg aaacatgaga gcttagtacg tactatcaac aggttgaact
     4381 gctgatcgat cttttctacg gggcggccgc gtcagtcagt catcatacca taacttcgta
     4441 tagcatacat tatacgaagt tatctggttt g
//
