LOCUS       V010339                10321 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V010339
VERSION     V010339
KEYWORDS    pZIP-TRE3G-ZsGreen-Puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10321)
  AUTHORS   transOMIC
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 10321)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..10321
                     /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"
     polyA_signal    2187..2321
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     ncRNA           complement(2487..2613)
                     /label="3' miR-30a"
                     /note="sequence downstream of the 71-nt precursor of the
                     human miR-30a microRNA (Zeng et al., 2002)"
     misc_RNA        complement(2614..2682)
                     /label="shRNA"
                     /note="shRNA"
                     /note="short hairpin RNA"
     gap             2617..2638
                     /estimated_length=22
     gap             2658..2679
                     /estimated_length=22
     ncRNA           complement(2683..2804)
                     /label="5' miR-30a"
                     /note="sequence upstream of the 71-nt precursor of the
                     human miR-30a microRNA (Zeng et al., 2002)"
     CDS             complement(2882..3574)
                     /label="ZsGreen1"
                     /note="Zoanthus green fluorescent protein"
     promoter        complement(3614..3981)
                     /label="TRE3GS promoter"
                     /note="3rd-generation Tet-responsive promoter that can be
                     activated by binding of Tet-On(R) 3G, modified to eliminate
                     binding sites for endogenous mammalian transcription
                     factors"
     promoter        4002..4512
                     /label="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             4531..5274
                     /label="Tet-On(R) 3G"
                     /note="modified rtTA protein that binds tightly to
                     promoters containing the tet operator in the presence of
                     doxycycline"
     promoter        5288..5617
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             5626..6222
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    6243..6831
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             7039..7672
                     /label="5' LTR"
                     /note="5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(7849..7879)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7894..7915)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(8203..8788)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(8962..9819)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(9820..9924)
                     /label="AmpR promoter"
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 actacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgacttaact tgtttattgc agcttataat ggttacaaat
     2221 aaggcaatag catcacaaat ttcacaaata aggcattttt ttcactgcat tctagttttg
     2281 gtttgtccaa actcatcaat gtatcttatc atgtctggat ctcaaatccc tcggaagctg
     2341 cgcctgtctt aggttggagt gatacatttt tatcactttt acccgtcttt ggattaggca
     2401 gtagctctga cggccctcct gtcttaggtt agtgaaaaat gtcactctct tacccgtcat
     2461 tggctgtcca gacgcgtcaa ttgaaaaaag tgatttaatt tataccattt taattcagct
     2521 ttgtaaaaat gtatcaaaga gatagcaagg tattcagttt tagtaaacaa gataattgct
     2581 cctaaagtag ccccttgaag tccgaggcag taggcannnn nnnnnnnnnn nnnnnnnnta
     2641 catctgtggc ttcactannn nnnnnnnnnn nnnnnnnnnc gctcactgtc aacagcaata
     2701 taccttcttt agccttctgt tgggttaacc tgaagaagta atcccagcaa gtgtttccaa
     2761 gatgtgcagg caacgattct gtaaagtact gaagcctcat tcaaacaatt accctgttat
     2821 ccctagtcga gagatcttat catatgacta gtaaattcta gagtcgcggc cgcgatcttc
     2881 agggcaaggc ggagccggag gcgatggcgt gctcggtcag gtgccacttc tggttcttgg
     2941 cgtcgctgcg gtcctcgcgg gtcagcttgt gctggatgaa gtgccagtcg ggcatcttgc
     3001 ggggcacgga cttggccttg tacacggtgt cgaactggca gcgcaagcgg ccaccgtcct
     3061 tcagcagcag gtacatgctc acgtcgccct tcaagatgcc ctgcttgggc acggggatga
     3121 tcttctcgca ggagggctcc cagttgtcgg tcatcttctt catcacgggg ccgtcggcgg
     3181 ggaagttcac gccgtagaac ttggactcgt ggtacatgca gttctcctcc acgctcacgg
     3241 tgatgtcggc gttgcagatg cacacggcgc cgtcctcgaa caggaaggag cggtcccagg
     3301 tgtagccggc ggggcaggag ttcttgaagt agtcgacgat gtcctggggg tactcggtga
     3361 acacgcggtt gccgtacatg aaggcggcgg acaagatgtc ctcggcgaag ggcaaggggc
     3421 cgccctccac cacgcacagg ttgatggcct gcttgccctt gaaggggtag ccgatgccct
     3481 cgccggtgat cacgaacttg tggccgtcca cgcagccctc catgcggtac ttcatggtca
     3541 tctccttggt caggccgtgc ttggactggg ccatggtgga tccgccggca ccggtgtata
     3601 cgggaattaa ttctttacga gggtaggaag tggtacggaa agttggtata agacaaaagt
     3661 gttgtggaat tgaagtttac tcaaaaaatc agcactcttt tataggcgcc ctggtttaca
     3721 taagcaaagc ttatacgttc tctatcactg atagggagta aactggatat acgttctcta
     3781 tcactgatag ggagtaaact gtagatacgt tctctatcac tgatagggag taaactggtc
     3841 atacgttctc tatcactgat agggagtaaa ctccttatac gttctctatc actgataggg
     3901 agtaaagtct gcatacgttc tctatcactg atagggagta aactcttcat acgttctcta
     3961 tcactgatag ggagtaaact cgatcgaggt gataattcca cggggttggg gttgcgcctt
     4021 ttccaaggca gccctgggtt tgcgcaggga cgcggctgct ctgggcgtgg ttccgggaaa
     4081 cgcagcggcg ccgaccctgg gtctcgcaca ttcttcacgt ccgttcgcag cgtcacccgg
     4141 atcttcgccg ctacccttgt gggccccccg gcgacgcttc ctgctccgcc cctaagtcgg
     4201 gaaggttcct tgcggttcgc ggcgtgccgg acgtgacaaa cggaagccgc acgtctcact
     4261 agtaccctcg cagacggaca gcgccaggga gcaatggcag cgcgccgacc gcgatgggct
     4321 gtggccaata gcggctgctc agcagggcgc gccgagagca gcggccggga aggggcggtg
     4381 cgggaggcgg ggtgtggggc ggtagtgtgg gccctgttcc tgcccgcgcg gtgttccgca
     4441 ttctgcaagc ctccggagcg cacgtcggca gtcggctccc tcgttgaccg aatcaccgac
     4501 ctctctcccc agggggatca tcgaattacc atgtctagac tggacaagag caaagtcata
     4561 aactctgctc tggaattact caatggagtc ggtatcgaag gcctgacgac aaggaaactc
     4621 gctcaaaagc tgggagttga gcagcctacc ctgtactggc acgtgaagaa caagcgggcc
     4681 ctgctcgatg ccctgccaat cgagatgctg gacaggcatc atacccactc ctgccccctg
     4741 gaaggcgagt catggcaaga ctttctgcgg aacaacgcca agtcataccg ctgtgctctc
     4801 ctctcacatc gcgacggggc taaagtgcat ctcggcaccc gcccaacaga gaaacagtac
     4861 gaaaccctgg aaaatcagct cgcgttcctg tgtcagcaag gcttctccct ggagaacgca
     4921 ctgtacgctc tgtccgccgt gggccacttt acactgggct gcgtattgga ggaacaggag
     4981 catcaagtag caaaagagga aagagagaca cctaccaccg attctatgcc cccacttctg
     5041 aaacaagcaa ttgagctgtt cgaccggcag ggagccgaac ctgccttcct tttcggcctg
     5101 gaactaatca tatgtggcct ggagaaacag ctaaagtgcg aaagcggcgg gccgaccgac
     5161 gcccttgacg attttgactt agacatgctc ccagccgatg cccttgacga ctttgacctt
     5221 gatatgctgc ctgctgacgc tcttgacgat tttgaccttg acatgctccc cgggtaaacg
     5281 cgcgaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     5341 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     5401 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     5461 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     5521 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     5581 gtgaggaggc ttttttggag gcctaggctt ttgcaaaacg cgaccatgac cgagtacaag
     5641 cccacggtgc gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc
     5701 gcgttcgccg actaccccgc cacgcgccac accgtcgacc cggaccgcca catcgagcgg
     5761 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     5821 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     5881 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     5941 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     6001 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     6061 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     6121 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     6181 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaacgcg cgcgtctgga
     6241 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     6301 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     6361 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     6421 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     6481 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     6541 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     6601 tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc
     6661 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     6721 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     6781 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaatta
     6841 attctgcagt cgagacctag aaaaacatgg agcaatcaca agtagcaata cagcagctac
     6901 caatgctgat tgtgcctggc tagaagcaca agaggaggag gaggtgggtt tttccagtca
     6961 cacctcaggt acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
     7021 taaaagaaaa gaggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg
     7081 atctgtggat ctaccacaca caaggctact tccctgatta gcagaactac acaccagggc
     7141 caggggtcag atatccactg acctttggat ggtgctacaa gctagtacca gttgagccag
     7201 ataaggtaga agaggccaat aaaggagaga acaccagctt gttacaccct gtgagcctgc
     7261 atgggatgga tgacccggag agagaagtgt tagagtggag gtttgacagc cgcctagcat
     7321 ttcatcacgt ggcccgagag ctgcatccgg agtacttcaa gaactgctga tatcgagctt
     7381 gctacaaggg actttccgct ggggactttc cagggaggcg tggcctgggc gggactgggg
     7441 agtggcgagc cctcagatcc tgcatataag cagctgcttt ttgcctgtac tgggtctctc
     7501 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     7561 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     7621 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt
     7681 tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag
     7741 aggccttgac attgctagcg ttttaccgtc gacctctagc tagagcttgg cgtaatcatg
     7801 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     7861 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     7921 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     7981 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     8041 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     8101 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     8161 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     8221 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     8281 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     8341 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     8401 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     8461 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     8521 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     8581 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     8641 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     8701 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca
     8761 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     8821 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     8881 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     8941 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     9001 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     9061 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     9121 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     9181 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     9241 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     9301 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     9361 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     9421 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     9481 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     9541 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     9601 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     9661 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     9721 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     9781 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     9841 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     9901 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     9961 atcgggagat caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
    10021 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
    10081 tcaatgtatc ttatcatgtc tggatcaact ggataactca agctaaccaa aatcatccca
    10141 aacttcccac cccataccct attaccactg ccaattacct agtggtttca tttactctaa
    10201 acctgtgatt cctctgaatt attttcattt taaagaaatt gtatttgtta aatatgtact
    10261 acaaacttag tagtttttaa agaaattgta tttgttaaat atgtactaca aacttagtag
    10321 t
//
