LOCUS       62056_56        5082 bp DNA     circular SYN 17-NOV-2020
DEFINITION  Cloning vector (72)_P_EF1a-mRuby3, complete sequence.
ACCESSION   MT891324
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5082)
  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 5082)
  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 5082)
  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..5082
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..266
                     /note="misc_signal; INS"
                     /regulatory_class="other"
     polyA_signal    283..507
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        523..1704
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     CDS             1738..2448
                     /label=mRuby3
                     /note="monomeric red fluorescent protein derived from
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
     promoter        complement(2470..2488)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    2649..2770
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      2860..3149
                     /note="misc_signal; INS"
                     /regulatory_class="other"
     rep_origin      complement(3237..3825)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3999..4856)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4857..4961)
                     /label=AmpR promoter
ORIGIN
        1 gtcttctcca tcggatcctg atcaggggag ctcacgggga cagccccccc ccaaagcccc
       61 cagggatgta attacgtccc tcccccgcta gggggcagca gcgaccgccc ggggctccgc
      121 tccggtccgg cgctcccccc gcatcccgag ccggcagcgt gcggggacag cccgggcacg
      181 gggaaggtgg cacgggatcg ctttcctctg aacgcttctc gctgctcttt gagcctgcag
      241 acacctgggg gatacgggga aaaccgcggc atatggcctc gactgtgcct tctagttgcc
      301 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
      361 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
      421 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
      481 atgctgggga tgcggtgggc tctatggctt ctgagggcta gcgctccggt gcccgtcagt
      541 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa
      601 ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
      661 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc
      721 tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc
      781 ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacg cccctggctg
      841 cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct
      901 tgcgcttaag gagccccttc gcctcgtgct tgagttgagg cctggcctgg gcgctggggc
      961 cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga taagtctcta
     1021 gccatttaaa atttttgatg acctgctgcg acgctttttt tctggcaaga tagtcttgta
     1081 aatgcgggcc aagatctgca cactggtatt tcggtttttg gggccgcggg cggcgacggg
     1141 gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga
     1201 atcggacggg ggtagtctca agctggccgg cctgctctgg tgcctggcct cgcgccgccg
     1261 tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa
     1321 agatggccgc ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga
     1381 gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc agccgtcgct
     1441 tcatgtgact ccacggagta ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt
     1501 tggagtacgt cgtctttagg ttggggggag gggttttatg cgatggagtt tccccacact
     1561 gagtgggtgg agactgaagt taggccagct tggcacttga tgtaattctc cttggaattt
     1621 gccctttttg agtttggatc ttggttcatt ctcaagcctc agacagtggt tcaaagtttt
     1681 tttcttccat ttcaggtgtc gtgacataaa ctgccctaga ttagcggccg cgccaccatg
     1741 gtgtctaagg gcgaagagct gatcaaggaa aatatgcgta tgaaggtggt catggaaggt
     1801 tcggtcaacg gccaccaatt caaatgcaca ggtgaaggag aaggcagacc gtacgaggga
     1861 gtgcaaacca tgaggatcaa agtcatcgag ggaggacccc tgccatttgc ctttgacatt
     1921 cttgccacgt cgttcatgta tggcagccgt acctttatca agtacccggc cgacatccct
     1981 gatttcttta aacagtcctt tcctgagggt tttacttggg aaagagttac gagatacgaa
     2041 gatggtggag tcgtcaccgt cacgcaggac accagccttg aggatggcga gctcgtctac
     2101 aacgtcaagg tcagaggggt aaactttccc tccaatggtc ccgtgatgca gaagaagacc
     2161 aagggttggg agcctaatac agagatgatg tatccagcag atggtggtct gagaggatac
     2221 actgacatcg cactgaaagt tgatggtggt ggccatctgc actgcaactt cgtgacaact
     2281 tacaggtcaa aaaagaccgt cgggaacatc aagatgcccg gtgtccatgc cgttgatcac
     2341 cgcctggaaa ggatcgagga gagtgacaat gaaacctacg tagtgcaaag agaagtggca
     2401 gttgccaaat acagcaacct tggtggtggc atggacgagc tgtacaagta gtaatctaga
     2461 gggccctatt ctatagtgtc acctaaatgc tagagctcgc tgatcagtcc atagtcgacg
     2521 gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac
     2581 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     2641 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     2701 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     2761 atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     2821 tgttaaatca gctcattttt taaccaatag gccgaaatcg gggagctcac ggggacagcc
     2881 cccccccaaa gcccccaggg atgtaattac gtccctcccc cgctaggggg cagcagcgac
     2941 cgcccggggc tccgctccgg tccggcgctc cccccgcatc ccgagccggc agcgtgcggg
     3001 gacagcccgg gcacggggaa ggtggcacgg gatcgctttc ctctgaacgc ttctcgctgc
     3061 tctttgagcc tgcagacacc tgggggatac ggggaaaatg atcaagatct tctacactgg
     3121 gacgcaagga ttatcctcga ggcgcgcgaa gaatcagggg ataacgcagg aaagaacatg
     3181 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3241 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3301 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3361 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3421 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3481 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3541 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3601 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3661 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3721 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3781 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3841 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3901 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     3961 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4021 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     4081 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     4141 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     4201 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     4261 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     4321 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     4381 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     4441 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     4501 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     4561 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     4621 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     4681 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     4741 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     4801 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     4861 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     4921 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     4981 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
     5041 tctgctctga tgccgcatag ttaagccagt atctgctccc tg
//
