LOCUS       V010347                 8959 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V010347
VERSION     V010347
KEYWORDS    pZIP-hEF1-alpha-RFP-Puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8959)
  AUTHORS   transOMIC
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8959)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8959
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        27..404
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     LTR             411..668
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    715..840
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1337..1570
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             1755..1799
                     /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             1948..1989
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    2061..2178
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2226..3406
                     /label="EF-1-alpha promoter"
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     CDS             3419..4111
                     /label="TurboRFP"
                     /note="red fluorescent protein from Entacmaea quadricolor"
     misc_feature    4128..4710
                     /label="IRES2"
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
     CDS             4709..5302
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     ncRNA           5326..5447
                     /label="5' miR-30a"
                     /note="sequence upstream of the 71-nt precursor of the
                     human miR-30a microRNA (Zeng et al., 2002)"
     misc_RNA        5448..5516
                     /label="shRNA"
                     /note="shRNA"
                     /note="short hairpin RNA"
     gap             5451..5472
                     /estimated_length=22
     gap             5492..5513
                     /estimated_length=22
     ncRNA           5517..5643
                     /label="3' miR-30a"
                     /note="sequence downstream of the 71-nt precursor of the
                     human miR-30a microRNA (Zeng et al., 2002)"
     misc_feature    5664..6252
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             6460..6693
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        6789..7118
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      complement(7207..7792)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7966..8823)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(8824..8928)
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatcctcg cgcgttgaca ttgattattg actagttatt aatagtaatc
       61 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      121 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      181 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      241 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      301 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      361 tcctacttgg cagtacatct acgtattagt catcgctatt accagctagc gaggcgtggc
      421 ctgggcggga ctggggagtg gcgagccctc agatcctgca tataagcagc tgctttttgc
      481 ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
      541 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
      601 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
      661 ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct
      721 cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg
      781 agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc
      841 agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga
      901 aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca
      961 gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa
     1021 ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc
     1081 tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag
     1141 gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccggccgctg atcttcagac
     1201 ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa
     1261 aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa
     1321 aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta
     1381 tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc
     1441 agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag
     1501 tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc
     1561 aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt
     1621 ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg
     1681 agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc
     1741 aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt
     1801 ggaattggtt taacataaca aattggctgt ggtatataaa attattcata atgatagtag
     1861 gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc
     1921 agggatattc accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc
     1981 ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga
     2041 acggatctcg acggtatcgc ctttaaaaga aaagggggga ttggggggta cagtgcaggg
     2101 gaaagaatag tagacataat agcaacagac atacaaacta aagaactaca aaaacaaatt
     2161 acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt ttatcgatac
     2221 gtgaggctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt
     2281 ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg taaactggga
     2341 aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag
     2401 tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggtaag
     2461 tgccgtgtgt ggttcccgcg ggcctggcct ctttacgggt tatggccctt gcgtgccttg
     2521 aattacttcc acgcccctgg ctgcagtacg tgattcttga tcccgagctt cgggttggaa
     2581 gtgggtggga gagttcgagg ccttgcgctt aaggagcccc ttcgcctcgt gcttgagttg
     2641 aggcctggct tgggcgctgg ggccgccgcg tgcgaatctg gtggcacctt cgcgcctgtc
     2701 tcgctgcttt cgataagtct ctagccattt aaaatttttg atgacctgct gcgacgcttt
     2761 ttttctggca agatagtctt gtaaatgcgg gccaagatct gcacactggt atttcggttt
     2821 ttggggccgc gggcggcgac ggggcccgtg cgtcccagcg cacatgttcg gcgaggcggg
     2881 gcctgcgagc gcggccaccg agaatcggac gggggtagtc tcaagctggc cggcctgctc
     2941 tggtgcctgg cctcgcgccg ccgtgtatcg ccccgccctg ggcggcaagg ctggcccggt
     3001 cggcaccagt tgcgtgagcg gaaagatggc cgcttcccgg ccctgctgca gggagctcaa
     3061 aatggaggac gcggcgctcg ggagagcggg cgggtgagtc acccacacaa aggaaaaggg
     3121 cctttccgtc ctcagccgtc gcttcatgtg actccacgga gtaccgggcg ccgtccaggc
     3181 acctcgatta gttctcgagc ttttggagta cgtcgtcttt aggttggggg gaggggtttt
     3241 atgcgatgga gtttccccac actgagtggg tggagactga agttaggcca gcttggcact
     3301 tgatgtaatt ctccttggaa tttgcccttt ttgagtttgg atcttggttc attctcaagc
     3361 ctcagacagt ggttcaaagt ttttttcttc catttcaggt gtcgtgcccg gtgccaccat
     3421 gagcgagctg atcaaggaga acatgcacat gaagctgtac atggagggca ccgtgaacaa
     3481 ccaccacttc aagtgcacat ccgagggcga aggcaagccc tacgagggca cccagaccat
     3541 gaagatcaag gtggtcgagg gcggccctct ccccttcgcc ttcgacatcc tggctaccag
     3601 cttcatgtac ggcagcaaag ccttcatcaa ccacacccag ggcatccccg acttctttaa
     3661 gcagtccttc cctgagggct tcacatggga gagaatcacc acatacgaag acgggggcgt
     3721 gctgaccgct acccaggaca ccagcttcca gaacggctgc atcatctaca acgtcaagat
     3781 caacggggtg aacttcccat ccaacggccc tgtgatgcag aagaaaacac gcggctggga
     3841 ggccaacacc gagatgctgt accccgctga cggcggcctg agaggccaca gccagatggc
     3901 cctgaagctc gtgggcgggg gctacctgca ctgctccttc aagaccacat acagatccaa
     3961 gaaacccgct aagaacctca agatgcccgg cttccacttc gtggaccaca gactggaaag
     4021 aatcaaggag gccgacaaag agacctacgt cgagcagcac gagatggctg tggccaagta
     4081 ctgcgacctc cctagcaaac tggggcacag ataagaattc aattccgccc ctctccctcc
     4141 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
     4201 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
     4261 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg
     4321 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta
     4381 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
     4441 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
     4501 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
     4561 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca
     4621 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     4681 ctttgaaaaa cacgataata ccatggccac cgagtacaag cccacggtgc gcctcgccac
     4741 ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     4801 cacgcgccac accgtcgacc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     4861 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     4921 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     4981 ctcgcgcatg gccaagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     5041 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     5101 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     5161 gcgcgctggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     5221 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     5281 catgacccgc aagcccggtg cctgagcggc cgcaatggat cctgtttgaa tgaggcttca
     5341 gtactttaca gaatcgttgc ctgcacatct tggaaacact tgctgggatt acttcttcag
     5401 gttaacccaa cagaaggcta aagaaggtat attgctgttg acagtgagcg nnnnnnnnnn
     5461 nnnnnnnnnn nntagtgaag ccacagatgt annnnnnnnn nnnnnnnnnn nnntgcctac
     5521 tgcctcggac ttcaaggggc tactttagga gcaattatct tgtttactaa aactgaatac
     5581 cttgctatct ctttgataca tttttacaaa gctgaattaa aatggtataa attaaatcac
     5641 tttacgcgtg gcgcgcctgg aacaatcaac ctctggatta caaaatttgt gaaagattga
     5701 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     5761 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     5821 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     5881 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     5941 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     6001 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc
     6061 tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     6121 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     6181 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     6241 ccgcctcccc gcctggaatt aattctgcag tcgagaccta gaaaaacatg gagcaatcac
     6301 aagtagcaat acagcagcta ccaatgctga ttgtgcctgg ctagaagcac aagaggagga
     6361 ggaggtgggt ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag
     6421 ctgtagatct tagccacttt ttaaaagaaa agaggggact ggaagggcta attcactccc
     6481 aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag
     6541 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     6601 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     6661 gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc
     6721 agtatttata acttgcaaag aaatgaatat cagagagtga gaggccttga cattgtttaa
     6781 actggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     6841 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     6901 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     6961 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     7021 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     7081 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     7141 ccattacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     7201 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     7261 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     7321 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     7381 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     7441 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     7501 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     7561 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     7621 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     7681 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     7741 cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     7801 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     7861 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     7921 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     7981 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     8041 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     8101 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     8161 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     8221 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     8281 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     8341 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     8401 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     8461 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     8521 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     8581 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     8641 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     8701 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     8761 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     8821 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     8881 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     8941 aaaagtgcca cctgacgtc
//
