LOCUS       62056_3850        4554 bp DNA     circular SYN 17-NOV-2020
DEFINITION  Cloning vector pBI-CMV-EGFP-mKate-1xmiR31ts3'UTR, complete sequence.
ACCESSION   MT891346
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4554)
  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 4554)
  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 4554)
  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..4554
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    complement(22..103)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(223..939)
                     /label=EGFP
                     /note="enhanced GFP"
     regulatory      complement(963..1031)
                     /label=minimal CMV promoter 2
                     /note="minimal CMV promoter 2"
                     /regulatory_class="promoter"
     enhancer        1098..1401
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1402..1605
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             1649..1666
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             1667..2374
                     /label=PAmKate
                     /note="monomeric photoactivatable red fluorescent protein 
                     (Gunewardene et al., 2011)"
     misc_feature    2384..2404
                     /label=miR31-TS
                     /note="miR31-TS"
     polyA_signal    2556..2637
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2819..3407)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3581..4438)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4439..4543)
                     /label=AmpR promoter
ORIGIN
        1 gaaaagtgcc acctgacgtc ggcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
       61 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
      121 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
      181 ctacaaatgt ggtatggctg attatgatcc tctagactgc agcttgtaca gctcgtccat
      241 gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc
      301 gttggggtct ttgctcaggg cggactgggt gctcaggtag tggttgtcgg gcagcagcac
      361 ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc
      421 ctcgatgttg tggcggatct tgaagttcac cttgatgccg ttcttctgct tgtcggccat
      481 gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc
      541 ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt
      601 cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac
      661 gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg
      721 gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg gcacgggcag
      781 cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat cgccctcgcc
      841 ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg
      901 caccaccccg gtgaacagct cctcgccctt gctcaccatg gtggcaccgg tgaattctcc
      961 aggcgatctg acggttcact aaacgagctc tgcttatata ggcctcccac cgtacacgcc
     1021 acctcgacat actcgagtag ttattaatag taatcaatta cggggtcatt agttcatagc
     1081 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     1141 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     1201 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     1261 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     1321 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     1381 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
     1441 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
     1501 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
     1561 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt
     1621 cagatccgct agggatcccg ccaccatgca ccatcaccat caccatgtgt ctaagggcga
     1681 agagctgatt aaggagaaca tgcacatgaa gctgtacatg gagggcaccg tgaacaacca
     1741 ccacttcaag tgcacatccg agggcgaagg caagccctac gagggcaccc agaccatgag
     1801 aatcaaggtg gtcgagggcg gccctctccc cttcgccttc gacatcctgg ctaccagctt
     1861 catgtacggc agcaaaacct tcatcaacca cacccagggc atccccgact tctttaagca
     1921 gtccttccct gagggcttca catgggagag agtcaccaca tacgaagacg ggggcgtgct
     1981 gaccgctacc caggacacca gcctccagga cggctgcctc atctacaacg tcaagatcag
     2041 aggggtgaac ttcccatcca acggccctgt gatgcagaag aaaacactcg gctgggaggc
     2101 ctccaccgag atgctgtacc ccgctgacgg cggcctggaa ggcagaagcg acatggccct
     2161 gaagctcgtg ggcgggggcc acctgatctg caacttgaag accacataca gatccaagaa
     2221 acccgctaag aacctcaaga tgcccggcgt ctactatgtg gacagaagac tggaaagaat
     2281 caaggaggcc gacaaagaga cctacgtcga gcagcacgag gtggctgtgg ccagatactg
     2341 cgacctccct agcaaactgg ggcacaaact taattgaaag cttagctatg ccagcatctt
     2401 gcctttaatt aaagctagtt acccaagctt gtcgacgata tctccagagg atcataatca
     2461 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     2521 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     2581 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcccc
     2641 gagcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     2701 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2761 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2821 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2881 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2941 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3001 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3061 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3121 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3181 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3241 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3301 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3361 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3421 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3481 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3541 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3601 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3661 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3721 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3781 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3841 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3901 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     3961 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4021 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4081 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4141 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaacacggg
     4201 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4261 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4321 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4381 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4441 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4501 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt cccc
//
