LOCUS       62056_20345        4144 bp DNA     circular SYN 17-NOV-2020
DEFINITION  Cloning vector pT-GTW6-CMV-mKate_1xmiR21ts3'UTR, complete sequence.
ACCESSION   MT891357
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4144)
  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 4144)
  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 4144)
  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..4144
                     /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..1533
                     /label=1xmiR21-TS
                     /note="1xmiR21-TS"
     protein_bind    complement(1548..1572)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    1701..1822
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1829..2284)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2311..2415
                     /label=AmpR promoter
     CDS             2416..3273
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      3447..4035
                     /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 ctaaagctta gctagttacc cagctttctt
     1561 gtacaaagtg gtacgcgtga cggccgcgac tctagatcat aatcagccat accacatttg
     1621 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     1681 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     1741 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     1801 ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc
     1861 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     1921 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     1981 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     2041 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     2101 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     2161 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     2221 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     2281 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     2341 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     2401 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     2461 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     2521 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     2581 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     2641 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     2701 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     2761 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     2821 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     2881 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     2941 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     3001 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     3061 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     3121 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     3181 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     3241 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     3301 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     3361 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     3421 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     3481 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     3541 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     3601 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     3661 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     3721 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     3781 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     3841 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     3901 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     3961 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     4021 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     4081 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     4141 gcca
//
