LOCUS       62056_51        4129 bp DNA     circular SYN 17-NOV-2020
DEFINITION  Cloning vector (57)_P_EFS-mCitrine, complete sequence.
ACCESSION   MT891323
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4129)
  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 4129)
  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 4129)
  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..4129
                     /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        529..740
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             779..1495
                     /label=mEYFP
                     /note="enhanced YFP with monomerizing A206K mutation
                     (Zacharias et al., 2002)"
     promoter        complement(1517..1535)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    1696..1817
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      1907..2196
                     /note="misc_signal; INS"
                     /regulatory_class="other"
     rep_origin      complement(2284..2872)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3046..3903)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(3904..4008)
                     /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 gcatcgatgg gcagagcgca
      541 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc ggtgcctaga
      601 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg
      661 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg
      721 ggtttgccgc cagaacacag cataaactgc cctagattaa ttaagcggcc gcgccaccat
      781 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
      841 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
      901 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
      961 cgtgaccacc ttcggctacg gcctgatgtg cttcgcccgc taccccgacc acatgaagca
     1021 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     1081 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     1141 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     1201 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     1261 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     1321 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     1381 cctgagctac cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     1441 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtagta
     1501 atctagaggg ccctattcta tagtgtcacc taaatgctag agctcgctga tcagtccata
     1561 gtcgacggat ccaccggatc tagataactg atcataatca gccataccac atttgtagag
     1621 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     1681 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     1741 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     1801 ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
     1861 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcgggg agctcacggg
     1921 gacagccccc ccccaaagcc cccagggatg taattacgtc cctcccccgc tagggggcag
     1981 cagcgaccgc ccggggctcc gctccggtcc ggcgctcccc ccgcatcccg agccggcagc
     2041 gtgcggggac agcccgggca cggggaaggt ggcacgggat cgctttcctc tgaacgcttc
     2101 tcgctgctct ttgagcctgc agacacctgg gggatacggg gaaaatgatc aagatcttct
     2161 acactgggac gcaaggatta tcctcgaggc gcgcgaagaa tcaggggata acgcaggaaa
     2221 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2281 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2341 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2401 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2461 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2521 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2581 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2641 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2701 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     2761 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2821 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2881 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     2941 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3001 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     3061 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     3121 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     3181 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     3241 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     3301 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     3361 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     3421 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     3481 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3541 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     3601 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     3661 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3721 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3781 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3841 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     3901 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     3961 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     4021 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc
     4081 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctg
//
