LOCUS       62056_20370        4116 bp DNA     circular SYN 17-NOV-2020
DEFINITION  Cloning vector pT-GTW6-CMV-mKate_1xmiR31ts5'UTR, complete sequence.
ACCESSION   MT891348
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4116)
  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 4116)
  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 4116)
  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..4116
                     /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..758
                     /label=1xmiR31-TS
                     /note="1xmiR31-TS"
     CDS             772..1482
                     /label=PAmKate
                     /note="monomeric photoactivatable red fluorescent protein 
                     (Gunewardene et al., 2011)"
     CDS             1483..1500
                     /label=6xHis
                     /note="6xHis affinity tag"
     protein_bind    complement(1520..1544)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    1673..1794
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1801..2256)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2283..2387
                     /label=AmpR promoter
     CDS             2388..3245
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      3419..4007
                     /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 tggatccagc tatgccagca tcttgcctgc tagccgccac catggtgtct
      781 aagggcgaag agctgattaa ggagaacatg cacatgaagc tgtacatgga gggcaccgtg
      841 aacaaccacc acttcaagtg cacatccgag ggcgaaggca agccctacga gggcacccag
      901 accatgagaa tcaaggtggt cgagggcggc cctctcccct tcgccttcga catcctggct
      961 accagcttca tgtacggcag caaaaccttc atcaaccaca cccagggcat ccccgacttc
     1021 tttaagcagt ccttccctga gggcttcaca tgggagagag tcaccacata cgaagacggg
     1081 ggcgtgctga ccgctaccca ggacaccagc ctccaggacg gctgcctcat ctacaacgtc
     1141 aagatcagag gggtgaactt cccatccaac ggccctgtga tgcagaagaa aacactcggc
     1201 tgggaggcct ccaccgagat gctgtacccc gctgacggcg gcctggaagg cagaagcgac
     1261 atggccctga agctcgtggg cgggggccac ctgatctgca acttgaagac cacatacaga
     1321 tccaagaaac ccgctaagaa cctcaagatg cccggcgtct actatgtgga cagaagactg
     1381 gaaagaatca aggaggccga caaagagacc tacgtcgagc agcacgaggt ggctgtggcc
     1441 agatactgcg acctccctag caaactgggg cacaaactta atcaccatca ccatcaccat
     1501 tgattaatta aagctagtta cccagctttc ttgtacaaag tggtacgcgt gacggccgcg
     1561 actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     1621 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     1681 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     1741 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt
     1801 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     1861 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     1921 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     1981 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     2041 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     2101 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     2161 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     2221 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     2281 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     2341 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     2401 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     2461 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     2521 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     2581 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     2641 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     2701 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     2761 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     2821 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     2881 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     2941 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     3001 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     3061 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     3121 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     3181 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     3241 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     3301 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     3361 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3421 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3481 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3541 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     3601 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     3661 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     3721 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3781 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3841 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3901 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3961 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4021 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     4081 tatcccctga ttctgtggat aaccgtatta ccgcca
//
