LOCUS       62056_3870        4520 bp DNA     circular SYN 17-NOV-2020
DEFINITION  Cloning vector pBI-CMV-EGFP-mKate, complete sequence.
ACCESSION   MT891342
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4520)
  AUTHORS   Frei T, Cella F, Tedeschi F, Gutierrez J, Stan GB, Khammash M, 
            Siciliano V.
  TITLE     Characterization and mitigation of gene expression burden in 
            mammalian cells
  JOURNAL   Nat Commun 11 (1), 4641 (2020)
  PUBMED    32934213
REFERENCE   2  (bases 1 to 4520)
  AUTHORS   Frei T, Cella F, Tedeschi F, Gutierrez J, Stan G-B., Khammash M, 
            Siciliano V.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-AUG-2020) Synthetic and Systems Biology for 
            Biomedicine, Italian Institute of Technology-IIT, Largo Barsanti e 
            Martteucci 53, Naples, Campania 80125, Italy
REFERENCE   3  (bases 1 to 4520)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun"; date: "2020"; volume: "11"; issue: "1"; pages: "4641"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (15-AUG-2020) Synthetic and Systems Biology for Biomedicine, Italian
            Institute of Technology-IIT, Largo Barsanti e Martteucci 53, Naples,
            Campania 80125, Italy"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4520
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             213..230
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     polyA_signal    365..446
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(628..1216)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1390..2247)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2248..2352)
                     /label=AmpR promoter
     polyA_signal    complement(2385..2466)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(2586..3302)
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     enhancer        3461..3764
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3765..3968
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             join(4022..4520,1..212)
                     /codon_start=1
                     /label=PAmKate
                     /note="monomeric photoactivatable red fluorescent protein 
                     (Gunewardene et al., 2011)"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTEMLYPADGGLEGRSD
                     MALKLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
ORIGIN
        1 agctcgtggg cgggggccac ctgatctgca acttgaagac cacatacaga tccaagaaac
       61 ccgctaagaa cctcaagatg cccggcgtct actatgtgga cagaagactg gaaagaatca
      121 aggaggccga caaagagacc tacgtcgagc agcacgaggt ggctgtggcc agatactgcg
      181 acctccctag caaactgggg cacaaactta atcaccatca ccatcaccat tgaaagcttg
      241 tcgacgatat ctccagagga tcataatcag ccataccaca tttgtagagg ttttacttgc
      301 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
      361 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
      421 cacaaataaa gcattttttt cactgccccg agcttcctcg ctcactgact cgctgcgctc
      481 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
      541 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
      601 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
      661 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
      721 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
      781 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
      841 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
      901 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
      961 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     1021 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     1081 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     1141 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     1201 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     1261 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     1321 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     1381 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     1441 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     1501 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     1561 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     1621 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     1681 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     1741 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     1801 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     1861 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     1921 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     1981 gagttgctct tgcccggcgt caacacggga taataccgcg ccacatagca gaactttaaa
     2041 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     2101 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     2161 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     2221 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     2281 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     2341 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcggcagtg aaaaaaatgc
     2401 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     2461 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     2521 gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga tcctctagac
     2581 tgcagcttgt acagctcgtc catgccgaga gtgatcccgg cggcggtcac gaactccagc
     2641 aggaccatgt gatcgcgctt ctcgttgggg tctttgctca gggcggactg ggtgctcagg
     2701 tagtggttgt cgggcagcag cacggggccg tcgccgatgg gggtgttctg ctggtagtgg
     2761 tcggcgagct gcacgctgcc gtcctcgatg ttgtggcgga tcttgaagtt caccttgatg
     2821 ccgttcttct gcttgtcggc catgatatag acgttgtggc tgttgtagtt gtactccagc
     2881 ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc
     2941 accagggtgt cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg
     3001 aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc
     3061 tgcttcatgt ggtcggggta gcggctgaag cactgcacgc cgtaggtcag ggtggtcacg
     3121 agggtgggcc agggcacggg cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg
     3181 ccgtaggtgg catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg
     3241 ccgtccagct cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc
     3301 atggtggcac cggtgaattc tccaggcgat ctgacggttc actaaacgag ctctgcttat
     3361 ataggcctcc caccgtacac gccacctcga catactcgag tagttattaa tagtaatcaa
     3421 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     3481 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     3541 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     3601 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     3661 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     3721 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     3781 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     3841 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     3901 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     3961 gcagagctgg tttagtgaac cgtcagatcc gctagggatc ctttttttgc tagccgccac
     4021 catggtgtct aagggcgaag agctgattaa ggagaacatg cacatgaagc tgtacatgga
     4081 gggcaccgtg aacaaccacc acttcaagtg cacatccgag ggcgaaggca agccctacga
     4141 gggcacccag accatgagaa tcaaggtggt cgagggcggc cctctcccct tcgccttcga
     4201 catcctggct accagcttca tgtacggcag caaaaccttc atcaaccaca cccagggcat
     4261 ccccgacttc tttaagcagt ccttccctga gggcttcaca tgggagagag tcaccacata
     4321 cgaagacggg ggcgtgctga ccgctaccca ggacaccagc ctccaggacg gctgcctcat
     4381 ctacaacgtc aagatcagag gggtgaactt cccatccaac ggccctgtga tgcagaagaa
     4441 aacactcggc tgggaggcct ccaccgagat gctgtacccc gctgacggcg gcctggaagg
     4501 cagaagcgac atggccctga
//
