LOCUS       62056_20435        4210 bp DNA     circular SYN 17-NOV-2020
DEFINITION  Cloning vector pT-GTW6-CMV-mKate_4xmiR21ts3'UTR, complete sequence.
ACCESSION   MT891360
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4210)
  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 4210)
  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 4210)
  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..4210
                     /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..1599
                     /label=4xmiR21-TS
                     /note="4xmiR21-TS"
     protein_bind    complement(1614..1638)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    1767..1888
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1895..2350)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2377..2481
                     /label=AmpR promoter
     CDS             2482..3339
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      3513..4101
                     /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 taagctatca acatcagtct gataagctaa agcttagcta gttacccagc
     1621 tttcttgtac aaagtggtac gcgtgacggc cgcgactcta gatcataatc agccatacca
     1681 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     1741 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     1801 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     1861 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
     1921 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     1981 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     2041 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     2101 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     2161 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     2221 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     2281 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     2341 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     2401 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     2461 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     2521 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     2581 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     2641 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     2701 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     2761 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     2821 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     2881 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     2941 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     3001 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     3061 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     3121 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     3181 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     3241 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     3301 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     3361 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     3421 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     3481 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     3541 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     3601 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     3661 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     3721 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     3781 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     3841 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     3901 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     3961 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     4021 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     4081 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     4141 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     4201 attaccgcca
//
