LOCUS       62056_20405        4188 bp DNA     circular SYN 17-NOV-2020
DEFINITION  Cloning vector pT-GTW6-CMV-mKate_3xmiR21ts3'UTR, complete sequence.
ACCESSION   MT891359
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4188)
  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 4188)
  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 4188)
  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..4188
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    59..79
                     /label=attB4
                     /note="core recombination site for the Gateway(R) BP
                     reaction"
     enhancer        151..454
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        455..658
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     protein_bind    717..737
                     /label=attB1
                     /note="core recombination site for the Gateway(R) BP
                     reaction"
     CDS             775..792
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             793..1500
                     /label=PAmKate
                     /note="monomeric photoactivatable red fluorescent protein 
                     (Gunewardene et al., 2011)"
     misc_feature    1512..1577
                     /label=3xmiR21-TS
                     /note="3xmiR21-TS"
     protein_bind    complement(1592..1616)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    1745..1866
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1873..2328)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2355..2459
                     /label=AmpR promoter
     CDS             2460..3317
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      3491..4079
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 catttcaggt gatccagtta tttaatagtc aattacggca tatatctagc agtgatatca
       61 actttgtata gaaaagttgg ctccgaattc tagttattaa tagtaatcaa ttacggggtc
      121 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
      181 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      241 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      301 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      361 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      421 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
      481 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
      541 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
      601 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg
      661 tttagtgaac cgtcagatca gagctggttt agtgaaccgt cagatcgaat tcgacccaag
      721 tttgtacaaa gaagcaggct gattcttcca tttcaggtgg atcccgccac catgcaccat
      781 caccatcacc atgtgtctaa gggcgaagag ctgattaagg agaacatgca catgaagctg
      841 tacatggagg gcaccgtgaa caaccaccac ttcaagtgca catccgaggg cgaaggcaag
      901 ccctacgagg gcacccagac catgagaatc aaggtggtcg agggcggccc tctccccttc
      961 gccttcgaca tcctggctac cagcttcatg tacggcagca aaaccttcat caaccacacc
     1021 cagggcatcc ccgacttctt taagcagtcc ttccctgagg gcttcacatg ggagagagtc
     1081 accacatacg aagacggggg cgtgctgacc gctacccagg acaccagcct ccaggacggc
     1141 tgcctcatct acaacgtcaa gatcagaggg gtgaacttcc catccaacgg ccctgtgatg
     1201 cagaagaaaa cactcggctg ggaggcctcc accgagatgc tgtaccccgc tgacggcggc
     1261 ctggaaggca gaagcgacat ggccctgaag ctcgtgggcg ggggccacct gatctgcaac
     1321 ttgaagacca catacagatc caagaaaccc gctaagaacc tcaagatgcc cggcgtctac
     1381 tatgtggaca gaagactgga aagaatcaag gaggccgaca aagagaccta cgtcgagcag
     1441 cacgaggtgg ctgtggccag atactgcgac ctccctagca aactggggca caaacttaat
     1501 tgattaatta atcaacatca gtctgataag ctatcaacat cagtctgata agctatcaac
     1561 atcagtctga taagctaaag cttagctagt tacccagctt tcttgtacaa agtggtacgc
     1621 gtgacggccg cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc
     1681 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     1741 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     1801 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     1861 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     1921 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     1981 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     2041 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     2101 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     2161 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     2221 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     2281 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     2341 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     2401 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     2461 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     2521 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     2581 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     2641 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     2701 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     2761 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     2821 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     2881 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     2941 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     3001 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     3061 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     3121 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     3181 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     3241 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     3301 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     3361 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     3421 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     3481 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3541 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3601 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     3661 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3721 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3781 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3841 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3901 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3961 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     4021 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4081 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     4141 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgcca
//
