LOCUS       40924_40207        6728 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSI-AKR1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6728)
  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 6728)
  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 6728)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6728)
  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..6728
                     /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"
     protein_bind    2994..3041
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             3049..4068
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    4201..4334
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        4384..4488
                     /label=AmpR promoter
     promoter        4490..4847
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4882..5673
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5908..5955
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      complement(6048..6636)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    complement(6690..6723)
                     /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 ttaagatcga tccctaggct
     2941 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtac ctagaagttc
     3001 ctattccgaa gttcctattc tctagaaagt ataggaactt ccttggccaa aaagcctgaa
     3061 ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg
     3121 atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg agggcgtgga
     3181 tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg
     3241 cactttgcat cggccgcgct cccgattccg gaagtgcttg acattgggga attcagcgag
     3301 agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa
     3361 accgaactgc ccgctgttct gcagccggtc gcggaggcca tggatgcgat cgctgcggcc
     3421 gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg tcaatacact
     3481 acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg gcaaactgtg
     3541 atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat gctttgggcc
     3601 gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa caatgtcctg
     3661 acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt cggggattcc
     3721 caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat ggagcagcag
     3781 acgcgctact tcgagcggag gcatccggag cttgcaggat cgccgcggct ccgggcgtat
     3841 atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa tttcgatgat
     3901 gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg gactgtcggg
     3961 cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt agaagtactc
     4021 gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata gcacgtacta
     4081 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     4141 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     4201 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4261 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4321 tatcatgtct gtataccgtc gacctctagc cctgcaggag gtggcacttt tcggggaaat
     4381 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4441 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
     4501 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     4561 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     4621 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4681 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4741 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     4801 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
     4861 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     4921 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4981 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     5041 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     5101 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     5161 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     5221 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     5281 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     5341 gatcaggatg atctggacga ggaacatcag gggctcgcgc cagccgaact gttcgccagg
     5401 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     5461 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     5521 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga ggaacttggc
     5581 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     5641 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     5701 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     5761 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     5821 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
     5881 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
     5941 ttgggtcgtt tgttcataaa cgcatccctt aacgtgagtt taaagaacat gtgagcaaaa
     6001 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6061 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6121 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6181 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6241 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6301 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6361 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6421 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6481 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6541 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6601 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6661 gggcggccgc gtcagtcagt catcatacca taacttcgta tagcatacat tatacgaagt
     6721 tatctggt
//
