LOCUS       62056_11        5327 bp DNA     circular SYN 17-NOV-2020
DEFINITION  Cloning vector (138)_P_EF1a-miRFP670, complete sequence.
ACCESSION   MT891326
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5327)
  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 5327)
  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 5327)
  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..5327
                     /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..2682
                     /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(2715..2733)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    2894..3015
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      3105..3394
                     /note="misc_signal; INS"
                     /regulatory_class="other"
     rep_origin      complement(3482..4070)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4244..5101)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5102..5206)
                     /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 gtagcaggtc atgcctctgg cagccccgca ttcgggaccg cctctcattc gaattgcgaa
     1801 catgaagaga tccacctcgc cggctcgatc cagccgcatg gcgcgcttct ggtcgtcagc
     1861 gaacatgatc atcgcgtcat ccaggccagc gccaacgccg cggaatttct gaatctcgga
     1921 agcgtactcg gcgttccgct cgccgagatc gacggcgatc tgttgatcaa gatcctgccg
     1981 catctcgatc ccaccgccga aggcatgccg gtcgcggtgc gctgccggat cggcaatccc
     2041 tctacggagt actgcggtct gatgcatcgg cctccggaag gcgggctgat catcgaactc
     2101 gaacgtgccg gcccgtcgat cgatctgtca ggcacgctgg cgccggcgct ggagcggatc
     2161 cgcacggcgg gttcactgcg cgcgctgtgc gatgacaccg tgctgctgtt tcagcagtgc
     2221 accggctacg accgggtgat ggtgtatcgt ttcgatgagc aaggccacgg cctggtattc
     2281 tccgagtgcc atgtgcctgg gctcgaatcc tatttcggca accgctatcc gtcgtcgact
     2341 gtcccgcaga tggcgcggca gctgtacgtg cggcagcgcg tccgcgtgct ggtcgacgtc
     2401 acctatcagc cggtgccgct ggagccgcgg ctgtcgccgc tgaccgggcg cgatctcgac
     2461 atgtcgggct gcttcctgcg ctcgatgtcg ccgtgccatc tgcagttcct gaaggacatg
     2521 ggcgtgcgcg ccaccctggc ggtgtcgctg gtggtcggcg gcaagctgtg gggcctggtt
     2581 gtctgtcacc attatctgcc gcgcttcatc cgtttcgagc tgcgggcgat ctgcaaacgg
     2641 ctcgccgaaa ggatcgcgac gcggatcacc gcgcttgaga gctaatgata gggcgcgcct
     2701 ctagagggcc ctattctata gtgtcaccta aatgctagag ctcgctgatc agtccatagt
     2761 cgacggatcc accggatcta gataactgat cataatcagc cataccacat ttgtagaggt
     2821 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     2881 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     2941 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     3001 catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     3061 atttttgtta aatcagctca ttttttaacc aataggccga aatcggggag ctcacgggga
     3121 cagccccccc ccaaagcccc cagggatgta attacgtccc tcccccgcta gggggcagca
     3181 gcgaccgccc ggggctccgc tccggtccgg cgctcccccc gcatcccgag ccggcagcgt
     3241 gcggggacag cccgggcacg gggaaggtgg cacgggatcg ctttcctctg aacgcttctc
     3301 gctgctcttt gagcctgcag acacctgggg gatacgggga aaatgatcaa gatcttctac
     3361 actgggacgc aaggattatc ctcgaggcgc gcgaagaatc aggggataac gcaggaaaga
     3421 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3481 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3541 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3601 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3661 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3721 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3781 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3841 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3901 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     3961 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4021 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4081 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4141 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4201 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4261 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4321 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4381 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     4441 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4501 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4561 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4621 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4681 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4741 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     4801 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     4861 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4921 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4981 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5041 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5101 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     5161 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5221 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
     5281 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctg
//
