LOCUS       V014756                 8715 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014756
VERSION     V014756
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8715)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8715
                     /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             634..1341
                     /label="mCherry"
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     CDS             1396..1992
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    2018..2606
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             2678..2911
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2943..3167
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3213..3641
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     polyA_signal    3784..3917
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3954..3970)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(3978..3994)
                     /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(4002..4032)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4047..4068)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4356..4944)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5118..5975)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(5976..6080)
                     /label="AmpR promoter"
     enhancer        6346..6725
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6726..6928
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     LTR             6943..7123
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7170..7295
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    7788..8021
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             8206..8250
                     /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             8399..8440
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    8548..8665
                     /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 ttctagagcc accatggtga gcaagggcga ggaggataac
      661 atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc
      721 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
      781 aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag
      841 ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag
      901 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg
      961 gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg
     1021 cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag
     1081 gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg
     1141 ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag
     1201 aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac
     1261 aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc
     1321 ggcatggacg agctgtacaa ggagggcaga ggaagtcttc taacatgcgg tgacgtggag
     1381 gagaatcccg gccctatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     1441 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     1501 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     1561 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     1621 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     1681 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg
     1741 caccggccca aggagcccgc gtggttcctg gccaccgtcg gagtctcgcc cgaccaccag
     1801 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg
     1861 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
     1921 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
     1981 aagcccggtg cctgactcga gtacgcgtta agtcgacaat caacctctgg attacaaaat
     2041 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     2101 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     2161 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     2221 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     2281 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     2341 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     2401 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     2461 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     2521 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     2581 gatctccctt tgggccgcct ccccgcgtcg actttaagac caatgactta caaggcagct
     2641 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     2701 cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
     2761 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     2821 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     2881 cccttttagt cagtgtggaa aatctctagc agggcccgtt taaacccgct gatcagcctc
     2941 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     3001 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     3061 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     3121 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     3181 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     3241 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     3301 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3361 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3421 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3481 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3541 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     3601 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatcacgtg
     3661 ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     3721 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     3781 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     3841 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     3901 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     3961 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     4021 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     4081 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     4141 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4201 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4261 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4321 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4381 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4441 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4501 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4561 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4621 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4681 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4741 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4801 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4861 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4921 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4981 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5041 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5101 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5161 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5221 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5281 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5341 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5401 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5461 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5521 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5581 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5641 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5701 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5761 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5821 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5881 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     5941 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6001 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6061 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     6121 ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     6181 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
     6241 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta
     6301 ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga
     6361 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     6421 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     6481 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     6541 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     6601 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     6661 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     6721 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     6781 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     6841 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     6901 tacggtggga ggtctatata agcagcgcgt tttgcctgta ctgggtctct ctggttagac
     6961 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     7021 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     7081 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg
     7141 acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag
     7201 cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg
     7261 gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat
     7321 cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata
     7381 tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat
     7441 cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag
     7501 aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga
     7561 taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca
     7621 ccgcacagca agcggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg
     7681 agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc
     7741 aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc
     7801 cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta
     7861 caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt
     7921 gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga
     7981 atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct
     8041 ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg
     8101 gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca
     8161 agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa
     8221 ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg
     8281 tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt
     8341 gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc
     8401 cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga aggtggagag
     8461 agagacagag acagatccat tcgattagtg aacggatcgg cactgcgtgc gccaattctg
     8521 cagacaaatg gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag
     8581 tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag aattacaaaa
     8641 acaaattaca aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg
     8701 gttaattaag gtacc
//
