LOCUS       V014230                10753 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014230
VERSION     V014230
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10753)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10753
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        238..617
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        618..820
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     LTR             835..1015
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1680..1913
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             2098..2142
                     /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             2291..2332
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    2440..2557
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2608..2848
                     /label="U6 promoter"
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_RNA        4738..4813
                     /label="gRNA scaffold"
                     /note="guide RNA scaffold for the Streptococcus pyogenes
                     CRISPR/Cas9 system"
     promoter        4875..5086
                     /label="EF-1-alpha core promoter"
                     /note="core promoter for human elongation factor
                     EF-1-alpha"
     CDS             5111..5827
                     /label="EGFP"
                     /note="enhanced GFP"
     misc_feature    5845..6433
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             6505..6738
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6770..6994
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7040..7468
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        7482..7811
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     promoter        7859..7906
                     /label="EM7 promoter"
                     /note="synthetic bacterial promoter"
     CDS             7925..8296
                     /label="BleoR"
                     /note="antibiotic-binding protein"
     polyA_signal    8429..8562
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     promoter        complement(8647..8677)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(8692..8713)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(9001..9589)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(9763..10620)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(10621..10725)
                     /label="AmpR promoter"
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta aggtaccgag ggcctatttc ccatgattcc ttcatatttg
     2641 catatacgat acaaggctgt tagagagata attagaatta atttgactgt aaacacaaag
     2701 atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta
     2761 aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc
     2821 ttggctttat atatcttgtg gaaaggacga aacaccggag acggttgtaa atgagcacac
     2881 aaaatacaca tgctaaaata ttatattcta tgacctttat aaaatcaacc aaaatcttct
     2941 ttttaataac tttagtatca ataattagaa tttttatgtt cctttttgca aacttttaat
     3001 aaaaatgagc aaaataaaaa aacgctagtt ttagtaactc gcgttgtttt cttcaccttt
     3061 aataatagct actccaccac ttgttcctaa gcggtcagct cctgcttcaa tcattttttg
     3121 agcatcttca aatgttctaa ctccaccagc tgctttaact aaagcattgt ctttaacaac
     3181 tgacttcatt agtttaacat cttcaaatgt tgcacctgat tttgaaaatc ctgttgatgt
     3241 tttaacaaat tctaatccag cttcaacagc tatttcacaa gctttcatga tttcttcttt
     3301 tgttaataaa caattttcca taatacattt aacaacatgt gatccagctg ctttttttac
     3361 agctttcatg tcttctaaaa ctaattcata atttttgtct tttaatgcac caatatttaa
     3421 taccatatca atttctgttg caccatcttt aattgcttca gaaacttcga atgcttttgt
     3481 agctgttgtg catgcaccta gaggaaaacc tacaacattt gttattccta catttgtgcc
     3541 ttttaataat tctttacaat agcttgttca atatgaatta acacaaactg ttgcaaaatc
     3601 aaattcaatt gcttcatcac ataattgttt aatttcagct ttcgtagcat cttgttttaa
     3661 taatgtgtga tctatatatt tgtttagttt cattttttct cctatatatt catttttaat
     3721 tttaattctt taataatttc gtctacttta actttagcgt tttgaacaga ttcaccaaca
     3781 cctataaaat aaatttttag tttaggttca gttccacttg ggcgaacagc aaatcatgac
     3841 ttatcttcta aataaaattt tagtaagtct tgtcctggca tattatacat tccatcgatg
     3901 tagtcttcaa cattaacaac tttaagtcca gcaatttgag ttaagggtgt tgctctcaat
     3961 gatttcatta atggttcaat ttttaatttc ttttcttctg gtttaaaatt caagtttaaa
     4021 gtgaaagtgt aatatgcacc catttcttta aataaatctt ctaaatagtc tactaatgtt
     4081 ttattttgtt ttttataaaa tcaagcagcc tctgctatta atatagaagc ttgtattcca
     4141 tctttatctc tagctgagtc atcaattaca tatccataac tttcttcata agcaaaaaca
     4201 aaatttaatc cgttatcttc ttctttagca atttctctac ccattcattt aaatccagtt
     4261 aaagttttta caatattaac tccatatttt tcatgagcga ttctatcacc caaatcactt
     4321 gttacaaaac ttgaatatag agccggattt tttggaatgc tatttaagcg ttttagattt
     4381 gataattttc aatcaattaa aattggtcct gtttgatttc catctaatct tacaaaatga
     4441 ccatcatgtt ttattgccat tccaaatctg tcagcatctg ggtcattcat aataataata
     4501 tctgcatcat gtttaatacc atattcaagc ggtatttttc atgcaggatc aaattctgga
     4561 tttggattta caacattttt aaatgtttca tcttcaaatg catgctcttc aacctcaata
     4621 acgttatatc ctgattcacg taatattttt ggggtaaatt tagttcctgt tccattaact
     4681 gcgctaaaaa taatttttaa atctttttta gcttcttgct cttttttgta cgtctctgtt
     4741 ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc
     4801 accgagtcgg tgcttttttg aattcgctag ctaggtcttg aaaggagtgg gaattggctc
     4861 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg
     4921 ggtcggcaat tgatccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt
     4981 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt
     5041 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggacc ggttctagag
     5101 cgctgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     5161 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     5221 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     5281 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     5341 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     5401 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     5461 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     5521 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     5581 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     5641 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     5701 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     5761 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     5821 gtacaagtaa cgcgttaagt cgacaatcaa cctctggatt acaaaatttg tgaaagattg
     5881 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     5941 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     6001 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     6061 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     6121 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     6181 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     6241 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     6301 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     6361 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     6421 gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     6481 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
     6541 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     6601 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     6661 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     6721 tgtggaaaat ctctagcagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     6781 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     6841 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     6901 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     6961 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     7021 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     7081 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     7141 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     7201 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     7261 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     7321 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     7381 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     7441 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     7501 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     7561 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     7621 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     7681 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     7741 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     7801 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtgt
     7861 tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta
     7921 aaccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc
     7981 ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc
     8041 cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc
     8101 ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc
     8161 ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg agatcggcga
     8221 gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt
     8281 ggccgaggag caggactgac acgtgctacg agatttcgat tccaccgccg ccttctatga
     8341 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     8401 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     8461 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     8521 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     8581 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     8641 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     8701 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     8761 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     8821 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     8881 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     8941 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     9001 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     9061 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     9121 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     9181 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     9241 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     9301 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     9361 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     9421 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     9481 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     9541 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     9601 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     9661 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     9721 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     9781 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     9841 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     9901 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     9961 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
    10021 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
    10081 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
    10141 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
    10201 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
    10261 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
    10321 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
    10381 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
    10441 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
    10501 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
    10561 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
    10621 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
    10681 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
    10741 agtgccacct gac
//
