LOCUS       V012827                 9371 bp    DNA     circular SYN 06-NOV-2023
DEFINITION  Exported.
ACCESSION   V012827
VERSION     V012827
KEYWORDS    pUB_smFLAG_H2B_MS2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9371)
  AUTHORS   Morisaki T, Lyon K, DeLuca KF, DeLuca JG, English BP, Zhang Z, Lavis
            LD, Grimm JB, Viswanathan S, Looger LL, Lionnet T, Stasevich TJ
  TITLE     Real-time quantification of single RNA translation dynamics in
            living cells.
  JOURNAL   Science. 2016 Jun 17;352(6292):1425-9. doi: 10.1126/science.aaf0899.
            Epub 2016 May 5.
   PUBMED   27313040
REFERENCE   2  (bases 1 to 9371)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi:
            "10.1126/science.aaf0899"; journalName: "Science"; date:
            "2016-06-17- 17"; volume: "352"; issue: "6292"; pages: "1425-9"
FEATURES             Location/Qualifiers
     source          1..9371
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label="3' LTR"
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1303..1536
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             1721..1765
                     /label="gp41 peptide"
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
                     al., 2013)"
     CDS             1914..1955
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    2058..2175
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2218..2616
                     /label="UbC promoter"
                     /note="human ubiquitin C promoter"
     regulatory      2622..2631
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2631..2654
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             2658..2681
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             2685..2708
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             3213..3236
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             3243..3266
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             3279..3302
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             3309..3332
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             3525..3548
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             3552..3575
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             3579..3602
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             3639..4013
                     /label="H2B"
                     /note="human histone H2B"
     misc_feature    5761..6349
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             6424..6657
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(6776..6794)
                     /label="pBRforEco"
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     promoter        6862..6966
                     /label="AmpR promoter"
     CDS             6967..7824
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      7998..8586
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     primer_bind     8740..8757
                     /label="L4440"
                     /note="L4440 vector, forward primer"
     primer_bind     8849..8868
                     /label="SV40pro-F"
                     /note="SV40 promoter/origin, forward primer"
     protein_bind    8997..9018
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        9033..9063
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    9071..9087
                     /label="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     9095..9111
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     primer_bind     9161..9180
                     /label="EBV-rev"
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtggc
     2221 ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct cctcacggcg agcgctgcca
     2281 cgtcagacga agggcgcagc gagcgtcctg atccttccgc ccggacgctc aggacagcgg
     2341 cccgctgctc ataagactcg gccttagaac cccagtatca gcagaaggac attttaggac
     2401 gggacttggg tgactctagg gcactggttt tctttccaga gagcggaaca ggcgaggaaa
     2461 agtagtccct tctcggcgat tctgcggagg gatctccgtg gggcggtgaa cgccgatgat
     2521 tatataagga cgcgccgggt gtggcacagc tagttccgtc gcagccggga tttgggtcgc
     2581 ggttcttgtt tgtggatcgc tgtgatcgtc acttggcggc cgccaccatg gactacaagg
     2641 acgacgacga caaaggtgac tacaaagatg atgacgataa aggcgactat aaggacgatg
     2701 acgacaaggg cggaaactca ctgatcaagg aaaacatgcg gatgaaggtg gtgatggagg
     2761 gctccgtgaa tggtcaccag ttcaagtgca ccggagaggg agagggaaac ccgtacatgg
     2821 gaactcagac catgcgcatt aaggtcatcg aaggaggtcc gctgccgttc gctttcgata
     2881 tcctggccac ttcgttcgga ggagggtcgc gcacgttcat caagtacccg aagggaatcc
     2941 cggacttctt taagcagtca ttcccggaag gattcacttg ggaacgggtg acccggtatg
     3001 aagatggagg tgtggtgact gtcatgcaag atacttcgct ggaggatggg tgcctcgtgt
     3061 accacgtcca agtccgcgga gtgaatttcc cgtccaacgg accagtgatg cagaaaaaga
     3121 cgaagggttg ggaacctaat actgaaatga tgtaccccgc agacggaggg ctgaggggct
     3181 acacccacat ggcgctgaag gtcgacggag gagattacaa ggatgacgac gataagcaac
     3241 aagattacaa agacgatgat gacaagggcc agcagggcga ctacaaggac gacgacgaca
     3301 agcagcagga ctacaaagat gacgatgata aaggaggagg acatctgtcc tgttcgttcg
     3361 tgaccaccta cagatcaaag aaaaccgtgg gaaacatcaa gatgccgggc attcatgccg
     3421 tcgaccaccg cctggagcgg ctcgaagaat cagacaatga gatgttcgtc gtgcaaagag
     3481 aacatgccgt ggccaagttc gcgggactgg gaggcggtgg aggcgattac aaagacgatg
     3541 atgacaaggg tgactataaa gacgacgatg acaaagggga ttacaaggat gatgatgata
     3601 agggaggcgg tggatcaggt ggaggaggtt cactgcagcc agagccagcg aagtctgctc
     3661 ccgccccgaa aaagggctcc aagaaggcgg tgactaaggc gcagaagaaa ggcggcaaga
     3721 agcgcaagcg cagccgcaag gagagctatt ccatctatgt gtacaaggtt ctgaagcagg
     3781 tccaccctga caccggcatt tcgtccaagg ccatgggcat catgaattcg tttgtgaacg
     3841 acattttcga gcgcatcgca ggtgaggctt cccgcctggc gcattacaac aagcgctcga
     3901 ccatcacctc cagggagatc cagacggccg tgcgcctgct gctgcctggg gagttggcca
     3961 agcacgccgt gtccgagggt actaaggcca tcaccaagta caccagcgct aagcatatgt
     4021 aggctagcta ggcggactgt tactgagctg cgttttacac cctttctttg acaaaaccta
     4081 acttgcgcag aaaaaaaaaa aataagagac aacattggca tggctttgtt tttttaaatt
     4141 ttttttaaag tttttttttt tttttttttt ttttaagttt ttttgttttg ttttggcgct
     4201 tttgactcag gatttaaaaa ctggaacggt gaaggcgaca gcagttggtt ggagcaaaca
     4261 tcccccaaag ttctacaaat gtggctgagg actttgtaca ttgttttgtt tttttttttt
     4321 ttggttttgt ctttttttaa tagtcattcc aagtatccat gaaataagtg gttacaggaa
     4381 gtccctcacc ctcccaaaag ccacccccac tcctaagagg aggatggtcg cgtccatgcc
     4441 ctgagtccac cccggggaag gtgacaaccg gtaacctaca aacgggtgga ggatcacccc
     4501 acccgacact tcacaatcaa ggggtacaat acacaagggt ggaggaacac cccaccctcc
     4561 agacacatta cacagaaatc caatcaaaca gaagcaccat cagggcttct gctaccaaat
     4621 ttatctcaaa aaactacaac aaggaatcac catcagggat tccctgtgca atatacgtca
     4681 aacgagggcc acgacgggag gacgatcacg cctcccgaat atcggcatgt ctggctttcg
     4741 aattcagtgc gtggagcatc agcccacgca gccaatcaga gtcgaataca agtcgacttt
     4801 cgcgaagagc atcagccttc gcgccattct tacacaaacc acactctccc ctacaggaac
     4861 agcatcagcg ttcctgccca gtacccaact caagaaaatt tatgtcccca tgcagcatca
     4921 gcgcatgggc cccaagaata catccccaac aaaatcacat ccgagcacca acagggctcg
     4981 gagtgttgtt tcttgtccaa ctggacaaac cctccatgga ccatcaggcc atggactctc
     5041 accaacaaga caaaaactac tcttctcgaa gcagcatcag cgcttcgaaa cactcgagca
     5101 tacattgtgc ctatttcttg ggtggacgat cacgccaccc atgctctcac gaatttcaaa
     5161 acacggacaa ggacgagcac caccagggct cgtcgttcca cgtccaatac gattacttac
     5221 ctttcgggat cacgatcacg gatcccgcag ctacatcact tccactcagg acattcaagc
     5281 atgcacgatc acggcatgct ccacaagtct caaccacaga aactaccaaa tgggttcagc
     5341 accagcgaac ccactcctac ctcaaacctc ttcccacaaa actggcaagc aggatcaccg
     5401 cttgcccatt ccaacatacc aaatcaaaaa caattactgg tacagcatca gcgtaccagc
     5461 ccacatctct cactactatc aaaaaccaaa ccgttcagca acagcgaacg gtacacacgg
     5521 aaaaatcaac tggtttacaa atacgaaaga cgatcacgct ttcgtccagc gcaaactatt
     5581 acgaaaaaca tccgacggga agagcaacag ccttcccgcg gcggaaaacc tcacaaaaac
     5641 acgacaaacg gatgcacgaa cacggcatcc gccgacaacc cacaaactta caaccaggca
     5701 aacggtgcag gatcaccgca ccgtacatca aacacctcag atctcatatc gatagatcct
     5761 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     5821 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     5881 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     5941 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     6001 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     6061 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     6121 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     6181 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     6241 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     6301 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tgagatcctt
     6361 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
     6421 gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt
     6481 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     6541 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     6601 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta
     6661 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga
     6721 gtgagaggcc cgggttaatt aaggaaaggg ctagatcatt cttgaagacg aaagggcctc
     6781 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     6841 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     6901 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     6961 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     7021 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     7081 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     7141 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     7201 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     7261 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     7321 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     7381 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     7441 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     7501 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     7561 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     7621 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     7681 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     7741 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     7801 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     7861 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     7921 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     7981 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     8041 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     8101 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     8161 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     8221 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     8281 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     8341 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     8401 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     8461 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     8521 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     8581 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     8641 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     8701 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     8761 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     8821 agcaagctca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     8881 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     8941 tccccgtggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     9001 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     9061 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga catgattacg
     9121 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     9181 aatgtatctt atcatgtctg gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa
     9241 cttcccaccc cataccctat taccactgcc aattacctgt ggtttcattt actctaaacc
     9301 tgtgattcct ctgaattatt ttcattttaa agaaattgta tttgttaaat atgtactaca
     9361 aacttagtag t
//
