LOCUS       62056_26        7342 bp DNA     circular SYN 17-NOV-2020
DEFINITION  Cloning vector (194)_P_EF1a-mCitrine-I, complete sequence.
ACCESSION   MT891329
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7342)
  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 7342)
  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 7342)
  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..7342
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..266
                     /note="misc_signal; INS"
                     /regulatory_class="other"
     polyA_signal    283..490
                     /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             1726..2442
                     /label=mEYFP
                     /note="enhanced YFP with monomerizing A206K mutation
                     (Zacharias et al., 2002)"
     regulatory      complement(2464..2482)
                     /label=SP6 promoter
                     /note="SP6 promoter"
                     /regulatory_class="promoter"
     polyA_signal    2643..2764
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      2854..3143
                     /note="misc_signal; INS"
                     /regulatory_class="other"
     polyA_signal    3156..3380
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        3415..3731
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3762..4706
                     /label=ORF frame 1
                     /note="bright monomeric near-infrared fluorescent protein 
                     engineered from a bacterial phytochrome, with an emission 
                     maximum of 670 nm (Shcherbakova et al., 2016)"
     promoter        complement(4739..4757)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     regulatory      4918..5039
                     /label=SV40 poly(A) signal
                     /note="SV40 poly(A) signal"
                     /regulatory_class="polyA_signal_sequence"
     regulatory      5129..5418
                     /note="misc_signal; INS"
                     /regulatory_class="other"
     rep_origin      complement(5497..6085)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6259..7116)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7117..7221)
                     /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 ccaccatggt gagcaagggc
     1741 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     1801 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     1861 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccttc
     1921 ggctacggcc tgatgtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
     1981 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     2041 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     2101 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     2161 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     2221 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     2281 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag
     2341 tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     2401 accgccgccg ggatcactct cggcatggac gagctgtaca agtagtaatc tagagggccc
     2461 tattctatag tgtcacctaa atgctagagc tcgctgatca gtccatagtc gacggatcca
     2521 ccggatctag ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     2581 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     2641 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     2701 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     2761 cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     2821 atcagctcat tttttaacca ataggccgaa atcggggagc tcacggggac agcccccccc
     2881 caaagccccc agggatgtaa ttacgtccct cccccgctag ggggcagcag cgaccgcccg
     2941 gggctccgct ccggtccggc gctccccccg catcccgagc cggcagcgtg cggggacagc
     3001 ccgggcacgg ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg
     3061 agcctgcaga cacctggggg atacggggaa aatgatcaag atcttctaca ctgggacgca
     3121 aggattatcc tcgaggcgcg cgaatatggc ctcgactgtg ccttctagtt gccagccatc
     3181 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     3241 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     3301 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     3361 ggatgcggtg ggctctatgg cttctgaggg ctagcgctcg cgagacgaaa gattggtgtg
     3421 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3481 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3541 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3601 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3661 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3721 gcttttgcaa acataaactg ccctagagcg gccgcgccac catggtagca ggtcatgcct
     3781 ctggcagccc cgcattcggg accgcctctc attcgaattg cgaacatgaa gagatccacc
     3841 tcgccggctc gatccagccg catggcgcgc ttctggtcgt cagcgaacat gatcatcgcg
     3901 tcatccaggc cagcgccaac gccgcggaat ttctgaatct cggaagcgta ctcggcgttc
     3961 cgctcgccga gatcgacggc gatctgttga tcaagatcct gccgcatctc gatcccaccg
     4021 ccgaaggcat gccggtcgcg gtgcgctgcc ggatcggcaa tccctctacg gagtactgcg
     4081 gtctgatgca tcggcctccg gaaggcgggc tgatcatcga actcgaacgt gccggcccgt
     4141 cgatcgatct gtcaggcacg ctggcgccgg cgctggagcg gatccgcacg gcgggttcac
     4201 tgcgcgcgct gtgcgatgac accgtgctgc tgtttcagca gtgcaccggc tacgaccggg
     4261 tgatggtgta tcgtttcgat gagcaaggcc acggcctggt attctccgag tgccatgtgc
     4321 ctgggctcga atcctatttc ggcaaccgct atccgtcgtc gactgtcccg cagatggcgc
     4381 ggcagctgta cgtgcggcag cgcgtccgcg tgctggtcga cgtcacctat cagccggtgc
     4441 cgctggagcc gcggctgtcg ccgctgaccg ggcgcgatct cgacatgtcg ggctgcttcc
     4501 tgcgctcgat gtcgccgtgc catctgcagt tcctgaagga catgggcgtg cgcgccaccc
     4561 tggcggtgtc gctggtggtc ggcggcaagc tgtggggcct ggttgtctgt caccattatc
     4621 tgccgcgctt catccgtttc gagctgcggg cgatctgcaa acggctcgcc gaaaggatcg
     4681 cgacgcggat caccgcgctt gagagctaat gatagggcgc gcctctagag ggccctattc
     4741 tatagtgtca cctaaatgct agagctcgct gatcagtcca tagtcgacgg atccaccgga
     4801 tctagataac tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     4861 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     4921 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     4981 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     5041 cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     5101 ctcatttttt aaccaatagg ccgaaatcgg ggagctcacg gggacagccc ccccccaaag
     5161 cccccaggga tgtaattacg tccctccccc gctagggggc agcagcgacc gcccggggct
     5221 ccgctccggt ccggcgctcc ccccgcatcc cgagccggca gcgtgcgggg acagcccggg
     5281 cacggggaag gtggcacggg atcgctttcc tctgaacgct tctcgctgct ctttgagcct
     5341 gcagacacct gggggatacg gggaaaatga tcaagatctt ctacactggg acgcaaggat
     5401 tatcctcgag gcgcgcgaag aatcagggga taacgcagtg tgagcaaaag gccagcaaaa
     5461 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5521 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5581 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5641 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5701 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5761 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5821 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5881 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     5941 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6001 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6061 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6121 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6181 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6241 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6301 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6361 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6421 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6481 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6541 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6601 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6661 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6721 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6781 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6841 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6901 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6961 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7021 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7081 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7141 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7201 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     7261 gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag
     7321 ttaagccagt atctgctccc tg
//
