LOCUS       V013232                 9532 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V013232
VERSION     V013232
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9532)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9532
                     /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    2027..2144
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2338..3519
                     /label="EF-1-alpha promoter"
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     CDS             3556..4230
                     /label="DsRed-Monomer"
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Strongin et al., 2007)"
     CDS             4297..4893
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     polyA_signal    4897..5104
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     promoter        5112..5352
                     /label="U6 promoter"
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_RNA        5377..5452
                     /label="gRNA scaffold"
                     /note="guide RNA scaffold for the Streptococcus pyogenes
                     CRISPR/Cas9 system"
     misc_feature    5453..6041
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             6248..6881
                     /label="5' LTR"
                     /note="5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(7010..7026)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(7034..7050)
                     /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(7058..7088)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7103..7124)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7412..8000)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(8174..9031)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(9032..9136)
                     /label="AmpR promoter"
     polyA_signal    9184..9318
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
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 tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgatgagtaa ttcatacaaa aggactcgcc cctgccttgg
     2221 ggaatcccag ggaccgtcgt taaactccca ctaacgtaga acccagagat cgctgcgttc
     2281 ccgccccctc acccgcccgc tctcgtcatc actgaggtgg agaagagcat gcgtgaggct
     2341 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag
     2401 gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg
     2461 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag
     2521 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt
     2581 gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt
     2641 ccacgcccct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
     2701 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
     2761 cttgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
     2821 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
     2881 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
     2941 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
     3001 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
     3061 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     3121 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     3181 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     3241 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     3301 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     3361 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     3421 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     3481 gtggttcaaa gtttttttct tccatttcag gtgtcgtgat tcgaattctg cagtcgacgg
     3541 taccgcgggc ccgggatgga caacaccgag gacgtcatca aggagttcat gcagttcaag
     3601 gtgcgcatgg agggctccgt gaacggccac tacttcgaga tcgagggcga gggcgagggc
     3661 aagccctacg agggcaccca gaccgccaag ctgcaggtga ccaagggcgg ccccctgccc
     3721 ttcgcctggg acatcctgtc cccccagttc cagtacggct ccaaggccta cgtgaagcac
     3781 cccgccgaca tccccgacta catgaagctg tccttccccg agggcttcac ctgggagcgc
     3841 tccatgaact tcgaggacgg cggcgtggtg gaggtgcagc aggactcctc cctgcaggac
     3901 ggcaccttca tctacaaggt gaagttcaag ggcgtgaact tccccgccga cggccccgta
     3961 atgcagaaga agactgccgg ctgggagccc tccaccgaga agctgtaccc ccaggacggc
     4021 gtgctgaagg gcgagatctc ccacgccctg aagctgaagg acggcggcca ctacacctgc
     4081 gacttcaaga ccgtgtacaa ggccaagaag cccgtgcagc tgcccggcaa ccactacgtg
     4141 gactccaagc tggacatcac caaccacaac gaggactaca ccgtggtgga gcagtacgag
     4201 cacgccgagg cccgccactc cggctcccag ggatcagagg gcagaggaag tcttctaaca
     4261 tgcggtgacg tggaggagaa tcccggccct tccgggatga ccgagtacaa gcccacggtg
     4321 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc
     4381 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag
     4441 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac
     4501 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc
     4561 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag
     4621 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc
     4681 ggcgtatcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg
     4741 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc
     4801 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg
     4861 cgcacctggt gcatgacccg caagcccggt gcctgactgt gccttctagt tgccagccat
     4921 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     4981 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     5041 ggggtggggt ggggcaggac agcaaggggg aggattggga agagaatagc aggcatgctg
     5101 gggaggtacc tgagggccta tttcccatga ttccttcata tttgcatata cgatacaagg
     5161 ctgttagaga gataattgga attaatttga ctgtaaacac aaagatatta gtacaaaata
     5221 cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa
     5281 tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct
     5341 tgtggaaagg acgaaacacc ggagacggac gtctcagttt tagagctaga aatagcaagt
     5401 taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gcaatcaacc
     5461 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     5521 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     5581 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     5641 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     5701 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     5761 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     5821 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     5881 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     5941 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     6001 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaatta attctgcagt
     6061 cgagacctag aaaaacatgg agcaatcaca agtagcaata cagcagctac caatgctgat
     6121 tgtgcctggc tagaagcaca agaggaggag gaggtgggtt ttccagtcac acctcaggta
     6181 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     6241 aggggactgg aagggctaat tcactcccaa cgaagacaag atatccttga tctgtggatc
     6301 taccacacac aaggctactt ccctgattag cagaactaca caccagggcc aggggtcaga
     6361 tatccactga cctttggatg gtgctacaag ctagtaccag ttgagccaga taaggtagaa
     6421 gaggccaata aaggagagaa caccagcttg ttacaccctg tgagcctgca tgggatggat
     6481 gacccggaga gagaagtgtt agagtggagg tttgacagcc gcctagcatt tcatcacgtg
     6541 gcccgagagc tgcatccgga gtacttcaag aactgctgat atcgagcttg ctacaaggga
     6601 ctttccgctg gggactttcc agggaggcgt ggcctgggcg ggactgggga gtggcgagcc
     6661 ctcagatcct gcatataagc agctgctttt tgcctgtact gggtctctct ggttagacca
     6721 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     6781 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     6841 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct
     6901 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggccttgaca
     6961 ttgctagcgt tttaccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     7021 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     7081 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     7141 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     7201 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     7261 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     7321 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     7381 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     7441 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     7501 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     7561 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     7621 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     7681 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     7741 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     7801 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     7861 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     7921 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     7981 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     8041 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     8101 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     8161 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     8221 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     8281 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     8341 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     8401 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     8461 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     8521 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     8581 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     8641 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     8701 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     8761 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     8821 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     8881 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     8941 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     9001 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     9061 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     9121 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     9181 atcaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     9241 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     9301 tcttatcatg tctggatcaa ctggataact caagctaacc aaaatcatcc caaacttccc
     9361 accccatacc ctattaccac tgccaattac ctgtggtttc atttactcta aacctgtgat
     9421 tcctctgaat tattttcatt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta
     9481 gtagttttta aagaaattgt atttgttaaa tatgtactac aaacttagta gt
//
