LOCUS       40924_40217        6408 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSI-AKR8, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6408)
  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 6408)
  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 6408)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6408)
  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..6408
                     /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=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     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"
     promoter        2934..3264
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3115..3250
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3312..3359
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             3378..3749
                     /codon_start=1
                     /label=BleoR
                     /note="antibiotic-binding protein"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    3882..4003
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        4064..4168
                     /label=AmpR promoter
     promoter        4170..4527
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4562..5353
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5588..5635
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      complement(5728..6316)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    complement(6370..6403)
                     /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 gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc
      781 gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc
      841 cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg
      901 tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac
      961 tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac
     1021 ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag
     1081 gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg
     1141 ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc
     1201 aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac
     1261 aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc
     1321 acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac
     1381 tccaccggcg gcatggacga gctgtacaag gctcaaagga gatataccat 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 ttaactgtgg aatgtgtgtc
     2941 agttagggtg tggaaagtcc ccaggctccc caggcaggca gaagtatgca aagcatgcat
     3001 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3061 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3121 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3181 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     3241 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     3301 gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca
     3361 aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt tccggtgctc accgcgcgcg
     3421 acgtcgccgg agcggtcgag ttctggaccg accggctcgg gttctcccgg gacttcgtgg
     3481 aggacgactt cgccggtgtg gtccgggacg acgtgaccct gttcatcagc gcggtccagg
     3541 accaggtggt gccggacaac accctggcct gggtgtgggt gcgcggcctg gacgagctgt
     3601 acgccgagtg gtcggaggtc gtgtccacga acttccggga cgcctccggg ccggccatga
     3661 ccgagatcgg cgagcagccg tgggggcggg agttcgccct gcgcgacccg gccggcaact
     3721 gcgtgcactt cgtggccgag gagcaggact gacacgtgct acgagatttc gattccaccg
     3781 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     3841 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     3901 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     3961 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgcctgcagg
     4021 aggtagggat aacagggtaa ttggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4081 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4141 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     4201 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4261 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     4321 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     4381 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4441 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     4501 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     4561 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     4621 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     4681 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     4741 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     4801 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     4861 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     4921 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     4981 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     5041 ggaacatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     5101 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     5161 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     5221 catagcgttg gctacccgtg atattgctga ggaacttggc ggcgaatggg ctgaccgctt
     5281 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     5341 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     5401 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     5461 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     5521 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     5581 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     5641 cgcatccctt aacgtgagtt taaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5701 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5761 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5821 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5881 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5941 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6001 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6061 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6121 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     6181 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6241 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6301 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggcggccgc gtcagtcagt
     6361 catcatacca taacttcgta tagcatacat tatacgaagt tatctggt
//
