LOCUS       V000415                10054 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V000415
VERSION     V000415
KEYWORDS    Tet-on circRNA Expression Vector
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10054)
  AUTHORS   Zhang Y, Xue W, Li X, Zhang J, Chen S, Zhang JL, Yang L, Chen LL
  TITLE     The Biogenesis of Nascent Circular RNAs.
  JOURNAL   Cell Rep. 2016 Apr 19;15(3):611-24. doi:
            10.1016/j.celrep.2016.03.058. Epub 2016 Apr 7.
   PUBMED   27068474
REFERENCE   2  (bases 1 to 10054)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 10054)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi:
            "10.1016/j.celrep.2016.03.058"; journalName: "Cell Rep"; date:
            "2016-04-19- 19"; volume: "15"; issue: "3"; pages: "611-24"
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..10054
                     /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        2227..2541
                     /label="tight TRE promoter"
                     /note="Tet-responsive promoter PTight, consisting of seven
                     tet operator sequences followed by the minimal CMV
                     promoter"
     regulatory      2569..2578
                     /label="Kozak sequence"
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2620..2641)
                     /label="EGFP-N"
                     /note="EGFP, reverse primer"
     primer_bind     3942..3961
                     /label="mCherry-F"
                     /note="mCherry, forward primer"
     misc_feature    4284..4870
                     /label="IRES2"
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5890..5909)
                     /label="EXFP-R"
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6237..6258
                     /label="EGFP-C"
                     /note="EGFP, forward primer"
     misc_feature    6444..7032
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             7107..7340
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7459..7477)
                     /label="pBRforEco"
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     promoter        7545..7649
                     /label="AmpR promoter"
     CDS             7650..8507
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      8681..9269
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     primer_bind     9423..9440
                     /label="L4440"
                     /note="L4440 vector, forward primer"
     primer_bind     9532..9551
                     /label="SV40pro-F"
                     /note="SV40 promoter/origin, forward primer"
     protein_bind    9680..9701
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        9716..9746
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    9754..9770
                     /label="lac operator"
                     /bound_moiety="lac repressor encoded by lacI"
                     /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     9759..9781
                     /label="M13/pUC Reverse"
                     /note="In lacZ gene"
     primer_bind     9778..9794
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     primer_bind     9778..9794
                     /label="M13 Reverse"
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9844..9863
                     /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 gactagttct
     2221 tcactcaagt ttactcccta tcagtgatag agaacgtatg tcgagtttac tccctatcag
     2281 tgatagagaa cgatgtcgag tttactccct atcagtgata gagaacgtat gtcgagttta
     2341 ctccctatca gtgatagaga acgtatgtcg agtttactcc ctatcagtga tagagaacgt
     2401 atgtcgagtt tatccctatc agtgatagag aacgtatgtc gagtttactc cctatcagtg
     2461 atagagaacg tatgtcgagg taggcgtgta cggtgggagg cctatataag cagagctcgt
     2521 ttagtgaacc gtcagatcgc ctggagaaaa gcgctagcgc taccggtcgc caccatggtg
     2581 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     2641 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     2701 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     2761 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     2821 gacttcttca agtccgccat gcccgaaggc tacgtccagg cacgtagcag gccagggcct
     2881 ctctcagcca cctgagcaga aagctttcca agatagggca ggctgggtta ggccatctga
     2941 gtctgtctcg ttcattggga tccagacttg actgtcttgt taaaggctgt tgctgcccag
     3001 gtgtgcaggg agctgttggt ctctggcatt cagggtgggg gtggtataaa cccggggcag
     3061 cttgcatatg gcagggaaga gggatccgtg gaggaacagt gcagaaggct ttatgttcag
     3121 aatctctctt gcttttcttc tagactgagt tccttgagat tggtgaatgc tgtgtattat
     3181 tcatccctga taacctggtg tttggcccag ggccttgtcc agaggagtgt ttgataagtg
     3241 tttcaagtga attagcacca cgatgtcatc tcttttcagt ttacaaagga cggacacccc
     3301 ctgacccggt ctcagaaagc ctgaaagcag aattagtcat tagaagggtg gttggcttgg
     3361 tcggcataga ctttgagcag aaagaggttg aaaatgttga gcctgatttc tcttaggccc
     3421 ctctgcagtg tctgttgtgg aggccagata cgtaactgct tccgcttttt ttggtgtcat
     3481 tcaaggtgag caaatcccct tcatgtttct caccagacaa tgcagctgat gaggttccag
     3541 ctttgcaaat gtagtcatcc atgaggactg tcttcctgag atttcatcag gctcgagtgg
     3601 acttgcaaag gactttaggt ccattgtcct tttattctta gatacctctt tcactgagac
     3661 cttttcctta cctcacctct ctaggtgacc aagggtggcc ccctgccctt cgcctgggac
     3721 atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc
     3781 cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
     3841 gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc
     3901 tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag
     3961 accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc
     4021 gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc
     4081 acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg
     4141 gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc
     4201 cgccactcca ccggcggcat ggacgagctg tacaaataag gtttagtgaa cggcgagatc
     4261 aacttcagat cttgataact cgagcctctc cctccccccc cccctaacgt tactggccga
     4321 agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg
     4381 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg
     4441 ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt
     4501 cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac
     4561 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca
     4621 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg
     4681 ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg
     4741 ggatctgatc tggggcctcg gtacacatgc tttacatgtg tttagtcgag gttaaaaaaa
     4801 cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat
     4861 ggccacaacc atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat
     4921 gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga
     4981 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtgt gtggtccaaa
     5041 tggaaacctg gtgggcagtg gggctctggg gtgcaaggtg gaggctagag aggaagagct
     5101 gtgttttttt tcctgactta cccagcagtg gtctgtgaga ttgtcttttc tggtgggcga
     5161 acaaaaaggg ggttaggaaa actcaggcca aaaaagtgta aggcgttaat tccccattta
     5221 attccttaaa atttcatgta ataccaggta ttgcctgtaa aggaaagata aagggaaaaa
     5281 taagtaagac cttgttaaaa ttttattttt ctattttaac cttcacttat ttcctaatta
     5341 ttaaaagaaa tttatgctta ttgttaagaa caaaaaaatt tcagtattac aatgaatttt
     5401 taattaaaag tttttggcct gatgaaatct caggaagaca gtcctcatgg atgactacat
     5461 ttgcaaagct ggaacctcat cagctgcatt gtctggtgag aaacatgaag gggatttgct
     5521 caccttgaat gagaccaaaa aaagcggaag cagttacgta tctggcctcc acaacagaca
     5581 ctgcagaggg gcctaagaga aatcaggctc aacattttca acctctttct gctcaaagtc
     5641 tatgccgacc aagccaacca cccttctaat gactaattct gctttcaggc tttctgagac
     5701 cgggtcaggg ggtgtcctta aaggttggaa aaaacttttc ctgtcatctt tgcctccaaa
     5761 atctggcttt ctcccttggg cagggaaacc tccccaacat ttctctatca tccctgagat
     5821 gtggggcctg cactctgact tctgtctgcc ttactctttg tcttacagga gcgcaccatc
     5881 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     5941 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     6001 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     6061 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     6121 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     6181 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     6241 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     6301 taaggatcca ccggatctag ataactgatc ataatcagcc ataccacatt tgtagaggtt
     6361 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     6421 attgttgttg ttcgatagat cctaatcaac ctctggatta caaaatttgt gaaagattga
     6481 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     6541 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     6601 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     6661 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     6721 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     6781 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     6841 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     6901 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     6961 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     7021 ccgcctcccc gcctgagatc ctttaagacc aatgacttac aaggcagctg tagatcttag
     7081 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga
     7141 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     7201 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     7261 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     7321 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact
     7381 tgcaaagaaa tgaatatcag agagtgagag gcccgggtta attaaggaaa gggctagatc
     7441 attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     7501 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     7561 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     7621 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     7681 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     7741 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     7801 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     7861 aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc
     7921 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     7981 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     8041 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     8101 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     8161 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     8221 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     8281 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     8341 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     8401 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     8461 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     8521 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     8581 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     8641 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     8701 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     8761 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     8821 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     8881 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     8941 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     9001 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     9061 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     9121 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     9181 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     9241 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     9301 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     9361 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     9421 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     9481 cgcgttggcc gattcattaa tgcagcaagc tcatggctga ctaatttttt ttatttatgc
     9541 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     9601 aggcctaggc ttttgcaaaa agctccccgt ggcacgacag gtttcccgac tggaaagcgg
     9661 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     9721 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     9781 gaaacagcta tgacatgatt acgaatttca caaataaagc atttttttca ctgcattcta
     9841 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcaac tggataactc
     9901 aagctaacca aaatcatccc aaacttccca ccccataccc tattaccact gccaattacc
     9961 tgtggtttca tttactctaa acctgtgatt cctctgaatt attttcattt taaagaaatt
    10021 gtatttgtta aatatgtact acaaacttag tagt
//
