LOCUS       62056_20365        4119 bp DNA     circular SYN 17-NOV-2020
DEFINITION  Cloning vector pT-GTW6-CMV-mKate_1xmiR31ts3'UTR, complete sequence.
ACCESSION   MT891351
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4119)
  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 4119)
  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 4119)
  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..4119
                     /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"
     CDS             748..765
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             766..1473
                     /label=PAmKate
                     /note="monomeric photoactivatable red fluorescent protein 
                     (Gunewardene et al., 2011)"
     misc_feature    1483..1503
                     /label=1xmiR31-TS
                     /note="1xmiR31-TS"
     protein_bind    complement(1523..1547)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    1676..1797
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1804..2259)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2286..2390
                     /label=AmpR promoter
     CDS             2391..3248
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      3422..4010
                     /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 tggatcccgc caccatgcac catcaccatc accatgtgtc taagggcgaa
      781 gagctgatta aggagaacat gcacatgaag ctgtacatgg agggcaccgt gaacaaccac
      841 cacttcaagt gcacatccga gggcgaaggc aagccctacg agggcaccca gaccatgaga
      901 atcaaggtgg tcgagggcgg ccctctcccc ttcgccttcg acatcctggc taccagcttc
      961 atgtacggca gcaaaacctt catcaaccac acccagggca tccccgactt ctttaagcag
     1021 tccttccctg agggcttcac atgggagaga gtcaccacat acgaagacgg gggcgtgctg
     1081 accgctaccc aggacaccag cctccaggac ggctgcctca tctacaacgt caagatcaga
     1141 ggggtgaact tcccatccaa cggccctgtg atgcagaaga aaacactcgg ctgggaggcc
     1201 tccaccgaga tgctgtaccc cgctgacggc ggcctggaag gcagaagcga catggccctg
     1261 aagctcgtgg gcgggggcca cctgatctgc aacttgaaga ccacatacag atccaagaaa
     1321 cccgctaaga acctcaagat gcccggcgtc tactatgtgg acagaagact ggaaagaatc
     1381 aaggaggccg acaaagagac ctacgtcgag cagcacgagg tggctgtggc cagatactgc
     1441 gacctcccta gcaaactggg gcacaaactt aattgaaagc ttagctatgc cagcatcttg
     1501 cctttaatta ataaagctag ttacccagct ttcttgtaca aagtggtacg cgtgacggcc
     1561 gcgactctag atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     1621 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     1681 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     1741 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg
     1801 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     1861 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     1921 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     1981 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     2041 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     2101 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     2161 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     2221 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     2281 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     2341 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     2401 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     2461 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     2521 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     2581 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     2641 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     2701 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     2761 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     2821 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     2881 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     2941 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     3001 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     3061 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     3121 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     3181 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     3241 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     3301 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     3361 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3421 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3481 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     3541 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     3601 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3661 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3721 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3781 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     3841 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     3901 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     3961 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4021 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     4081 cgttatcccc tgattctgtg gataaccgta ttaccgcca
//
