LOCUS       62056_20410        4161 bp DNA     circular SYN 17-NOV-2020
DEFINITION  Cloning vector pT-GTW6-CMV-mKate_3xmiR21ts5'UTR, complete sequence.
ACCESSION   MT891356
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4161)
  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 4161)
  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 4161)
  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..4161
                     /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    689..713
                     /label=attB1
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    738..803
                     /label=3xmiR21-TS
                     /note="3xmiR21-TS"
     CDS             817..1527
                     /label=PAmKate
                     /note="monomeric photoactivatable red fluorescent protein 
                     (Gunewardene et al., 2011)"
     CDS             1528..1545
                     /label=6xHis
                     /note="6xHis affinity tag"
     protein_bind    complement(1565..1589)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    1718..1839
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1846..2301)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2328..2432
                     /label=AmpR promoter
     CDS             2433..3290
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      3464..4052
                     /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 cgtcagatcg aattcgaccc aagtttgtac aaaaaagcag gctgattctt
      721 ccatttcagg tgctagctca acatcagtct gataagctat caacatcagt ctgataagct
      781 atcaacatca gtctgataag ctaggatccc gccaccatgg tgtctaaggg cgaagagctg
      841 attaaggaga acatgcacat gaagctgtac atggagggca ccgtgaacaa ccaccacttc
      901 aagtgcacat ccgagggcga aggcaagccc tacgagggca cccagaccat gagaatcaag
      961 gtggtcgagg gcggccctct ccccttcgcc ttcgacatcc tggctaccag cttcatgtac
     1021 ggcagcaaaa ccttcatcaa ccacacccag ggcatccccg acttctttaa gcagtccttc
     1081 cctgagggct tcacatggga gagagtcacc acatacgaag acgggggcgt gctgaccgct
     1141 acccaggaca ccagcctcca ggacggctgc ctcatctaca acgtcaagat cagaggggtg
     1201 aacttcccat ccaacggccc tgtgatgcag aagaaaacac tcggctggga ggcctccacc
     1261 gagatgctgt accccgctga cggcggcctg gaaggcagaa gcgacatggc cctgaagctc
     1321 gtgggcgggg gccacctgat ctgcaacttg aagaccacat acagatccaa gaaacccgct
     1381 aagaacctca agatgcccgg cgtctactat gtggacagaa gactggaaag aatcaaggag
     1441 gccgacaaag agacctacgt cgagcagcac gaggtggctg tggccagata ctgcgacctc
     1501 cctagcaaac tggggcacaa acttaatcac catcaccatc accattgatt aattaaagct
     1561 agttacccag ctttcttgta caaagtggta cgcgtgacgg ccgcgactct agatcataat
     1621 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     1681 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     1741 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     1801 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta
     1861 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     1921 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     1981 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     2041 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     2101 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     2161 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     2221 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     2281 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     2341 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2401 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     2461 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     2521 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     2581 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     2641 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     2701 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     2761 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     2821 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     2881 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     2941 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     3001 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     3061 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     3121 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     3181 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     3241 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     3301 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     3361 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     3421 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3481 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3541 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3601 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3661 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3721 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3781 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3841 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3901 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3961 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4021 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4081 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     4141 tggataaccg tattaccgcc a
//
