LOCUS       62056_20350        4117 bp DNA     circular SYN 17-NOV-2020
DEFINITION  Cloning vector pT-GTW6-CMV-mKate_1xmiR21ts5'UTR, complete sequence.
ACCESSION   MT891354
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4117)
  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 4117)
  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 4117)
  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..4117
                     /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..759
                     /label=3xmiR21-TS
                     /note="3xmiR21-TS"
     CDS             773..1483
                     /label=PAmKate
                     /note="monomeric photoactivatable red fluorescent protein 
                     (Gunewardene et al., 2011)"
     CDS             1484..1501
                     /label=6xHis
                     /note="6xHis affinity tag"
     protein_bind    complement(1521..1545)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    1674..1795
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1802..2257)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2284..2388
                     /label=AmpR promoter
     CDS             2389..3246
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      3420..4008
                     /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 gataagctag gatcccgcca ccatggtgtc
      781 taagggcgaa gagctgatta aggagaacat gcacatgaag ctgtacatgg agggcaccgt
      841 gaacaaccac cacttcaagt gcacatccga gggcgaaggc aagccctacg agggcaccca
      901 gaccatgaga atcaaggtgg tcgagggcgg ccctctcccc ttcgccttcg acatcctggc
      961 taccagcttc atgtacggca gcaaaacctt catcaaccac acccagggca tccccgactt
     1021 ctttaagcag tccttccctg agggcttcac atgggagaga gtcaccacat acgaagacgg
     1081 gggcgtgctg accgctaccc aggacaccag cctccaggac ggctgcctca tctacaacgt
     1141 caagatcaga ggggtgaact tcccatccaa cggccctgtg atgcagaaga aaacactcgg
     1201 ctgggaggcc tccaccgaga tgctgtaccc cgctgacggc ggcctggaag gcagaagcga
     1261 catggccctg aagctcgtgg gcgggggcca cctgatctgc aacttgaaga ccacatacag
     1321 atccaagaaa cccgctaaga acctcaagat gcccggcgtc tactatgtgg acagaagact
     1381 ggaaagaatc aaggaggccg acaaagagac ctacgtcgag cagcacgagg tggctgtggc
     1441 cagatactgc gacctcccta gcaaactggg gcacaaactt aatcaccatc accatcacca
     1501 ttgattaatt aaagctagtt acccagcttt cttgtacaaa gtggtacgcg tgacggccgc
     1561 gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     1621 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     1681 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     1741 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg
     1801 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     1861 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     1921 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     1981 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     2041 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     2101 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     2161 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     2221 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     2281 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2341 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     2401 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     2461 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     2521 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     2581 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     2641 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     2701 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     2761 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     2821 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     2881 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     2941 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     3001 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     3061 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     3121 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     3181 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     3241 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     3301 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     3361 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     3421 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     3481 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     3541 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     3601 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     3661 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     3721 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     3781 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     3841 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     3901 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     3961 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     4021 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     4081 ttatcccctg attctgtgga taaccgtatt accgcca
//
