LOCUS       V014865                 8889 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014865
VERSION     V014865
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8889)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8889
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..241
                     /label="U6 promoter"
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_RNA        266..341
                     /label="gRNA scaffold"
                     /note="guide RNA scaffold for the Streptococcus pyogenes
                     CRISPR/Cas9 system"
     promoter        403..614
                     /label="EF-1-alpha core promoter"
                     /note="core promoter for human elongation factor
                     EF-1-alpha"
     CDS             639..1355
                     /label="EGFP"
                     /note="enhanced GFP"
     misc_feature    1374..1962
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             2034..2267
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2299..2523
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2569..2997
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3011..3340
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     promoter        3388..3435
                     /label="EM7 promoter"
                     /note="synthetic bacterial promoter"
     CDS             3454..3825
                     /label="BleoR"
                     /note="antibiotic-binding protein"
     polyA_signal    3958..4091
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4128..4144)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(4152..4168)
                     /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)."
     promoter        complement(4176..4206)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4221..4242)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4530..5118)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5292..6149)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(6150..6254)
                     /label="AmpR promoter"
     enhancer        6520..6899
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6900..7102
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     LTR             7117..7297
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7344..7469
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    7962..8195
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             8380..8424
                     /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             8573..8614
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    8722..8839
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg gagacggacg tctctgtttt agagctagaa atagcaagtt aaaataaggc
      301 tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg cttttttgaa ttcgctagct
      361 aggtcttgaa aggagtggga attggctccg gtgcccgtca gtgggcagag cgcacatcgc
      421 ccacagtccc cgagaagttg gggggagggg tcggcaattg atccggtgcc tagagaaggt
      481 ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg
      541 ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg
      601 ccgccagaac acaggaccgg ttctagagcg ctgccaccat ggtgagcaag ggcgaggagc
      661 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
      721 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
      781 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
      841 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
      901 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
      961 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     1021 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     1081 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     1141 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     1201 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     1261 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     1321 ccgggatcac tctcggcatg gacgagctgt acaagtaaac gcgttaagtc gacaatcaac
     1381 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     1441 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     1501 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     1561 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     1621 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     1681 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     1741 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg
     1801 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     1861 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     1921 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcgtcgactt taagaccaat
     1981 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
     2041 gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt
     2101 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     2161 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     2221 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcaggg cccgtttaaa
     2281 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     2341 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     2401 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     2461 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     2521 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     2581 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     2641 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     2701 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     2761 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     2821 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     2881 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     2941 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     3001 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     3061 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     3121 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     3181 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3241 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     3301 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     3361 atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata gtatatcggc
     3421 atagtataat acgacaaggt gaggaactaa accatggcca agttgaccag tgccgttccg
     3481 gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg gctcgggttc
     3541 tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc
     3601 atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc
     3661 ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc
     3721 tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc
     3781 gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca cgtgctacga
     3841 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     3901 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     3961 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     4021 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     4081 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     4141 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     4201 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     4261 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     4321 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4381 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4441 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4501 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4561 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4621 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4681 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4741 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4801 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4861 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4921 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     4981 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5041 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5101 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5161 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5221 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5281 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5341 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5401 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5461 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5521 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5581 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5641 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5701 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5761 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5821 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5881 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5941 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6001 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6061 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6121 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6181 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6241 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
     6301 ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     6361 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
     6421 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
     6481 ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt
     6541 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     6601 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     6661 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     6721 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     6781 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     6841 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     6901 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     6961 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     7021 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     7081 gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc
     7141 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     7201 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     7261 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga
     7321 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca
     7381 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct
     7441 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat
     7501 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat
     7561 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag
     7621 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
     7681 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag
     7741 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac
     7801 agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
     7861 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
     7921 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
     7981 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
     8041 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
     8101 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
     8161 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
     8221 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
     8281 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
     8341 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
     8401 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
     8461 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
     8521 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
     8581 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac
     8641 agagacagat ccattcgatt agtgaacgga tcggcactgc gtgcgccaat tctgcagaca
     8701 aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg
     8761 ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat
     8821 tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccag tttggttaat
     8881 taaggtacc
//
