LOCUS       V010343                 9114 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V010343
VERSION     V010343
KEYWORDS    pZIP-mEF1-alpha-RFP-Puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9114)
  AUTHORS   transOMIC
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 9114)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9114
                     /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        2219..3562
                     /label="mEF-1-alpha promoter"
                     /note="strong constitutive promoter for mouse elongation
                     factor EF-1-alpha"
     CDS             3574..4266
                     /label="TurboRFP"
                     /note="red fluorescent protein from Entacmaea quadricolor"
     misc_feature    4283..4865
                     /label="IRES2"
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
     CDS             4864..5457
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     ncRNA           5481..5602
                     /label="5' miR-30a"
                     /note="sequence upstream of the 71-nt precursor of the
                     human miR-30a microRNA (Zeng et al., 2002)"
     misc_RNA        5603..5671
                     /label="shRNA"
                     /note="shRNA"
                     /note="short hairpin RNA"
     gap             5606..5627
                     /estimated_length=22
     gap             5647..5668
                     /estimated_length=22
     ncRNA           5672..5798
                     /label="3' miR-30a"
                     /note="sequence downstream of the 71-nt precursor of the
                     human miR-30a microRNA (Zeng et al., 2002)"
     misc_feature    5819..6407
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             6615..6848
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        6944..7273
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      complement(7362..7947)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(8121..8978)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(8979..9083)
                     /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 ttatcgatag
     2221 agtaattcat acaaaaggag ggatcgcctt cgcctgggga agtcccaggg accgtcgcta
     2281 aattctcata acccataatc ccggtacccg ccccaccaca gtgcgaggag catgcgctca
     2341 gggctgagcg cggggagagc agagcacaca agctcatagt ccctggtcgt ggggggggag
     2401 gggcgcactg agcggggggg ggggtgatgg ggggaggacc ggggagctgg cgcggggcaa
     2461 actgggaaag cggtgtcgtg tgctggctcc gccctcttcc cgagggtggg ggagaacggt
     2521 atataagtgc ggcagtcgcc ttggacgttc tttttcgcaa cgggtttgcc gtcaggacgc
     2581 aggtgagtgg cgggtgtggc ttccgcgggc cgccgagctg gaggtcctgc tccgagcggg
     2641 ccgggccccg ctgtcgtcgg cggggattag ctgcgagcat tcccgcttcg agttgcgggc
     2701 ggcgcgggag gcagagtgcg aggcctagcg gcaaccccgt agcctcgcct cgtgtccggc
     2761 ttgaggccta gcgtggtgtc cgcgccgccg ccgcgtgcta ctccggccgc actctggtct
     2821 tttttttttt gttgttgttg ccctgctgcc ttcgattgcc gttcagcaat aggggctaac
     2881 aaagggaggg tgcggggctt gctcacccgg agcccggaga ggtcatggtt ggggaggaat
     2941 ggagggacag gagtggcggc tggggcccgc ccgccttcgg agcacatgtc cgacgccacc
     3001 tggatggggc gaggcctggg gtttttcccg aagcaaccag gctcggggtt agcgttccga
     3061 ggccatgtgg ccccagcacc cggcacgatc tggcttggcg gcgccgcgtt gccctgcctc
     3121 cctaactagg gtgaggccat cccgtccggc accagttgcg tgcgcggaaa gatggccgct
     3181 cccgggccct gttgcaagga gctcaaaatg gaggacgcgg cagcccggtg gagcgggcgg
     3241 gtgagtcacc cacacaaagg aagagggcct ggtccctcac cggctgctgc ttcctgtgac
     3301 cccgtggtcc tatcggccgc aatagtcacc tcgggctttt gagcactgct cgtcgcggcg
     3361 gggggagggg atgtaatggc gttggagttt gttcacattt ggtgggtgga gactagtcag
     3421 gccagcctgg cgctggaagt cattcttgga atttgtcccc ttgagttttg agcggagcta
     3481 attctcgggc ttcttagcgg ttcaaaggta tcttttaaac ccttttttag gtgttgtgaa
     3541 aaccaccgct aattcaaagc aaccggtgcc accatgagcg agctgatcaa ggagaacatg
     3601 cacatgaagc tgtacatgga gggcaccgtg aacaaccacc acttcaagtg cacatccgag
     3661 ggcgaaggca agccctacga gggcacccag accatgaaga tcaaggtggt cgagggcggc
     3721 cctctcccct tcgccttcga catcctggct accagcttca tgtacggcag caaagccttc
     3781 atcaaccaca cccagggcat ccccgacttc tttaagcagt ccttccctga gggcttcaca
     3841 tgggagagaa tcaccacata cgaagacggg ggcgtgctga ccgctaccca ggacaccagc
     3901 ttccagaacg gctgcatcat ctacaacgtc aagatcaacg gggtgaactt cccatccaac
     3961 ggccctgtga tgcagaagaa aacacgcggc tgggaggcca acaccgagat gctgtacccc
     4021 gctgacggcg gcctgagagg ccacagccag atggccctga agctcgtggg cgggggctac
     4081 ctgcactgct ccttcaagac cacatacaga tccaagaaac ccgctaagaa cctcaagatg
     4141 cccggcttcc acttcgtgga ccacagactg gaaagaatca aggaggccga caaagagacc
     4201 tacgtcgagc agcacgagat ggctgtggcc aagtactgcg acctccctag caaactgggg
     4261 cacagataag aattcaattc cgcccctctc cctccccccc ccctaacgtt actggccgaa
     4321 gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt
     4381 cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg
     4441 gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc
     4501 ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc
     4561 ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa
     4621 aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc
     4681 tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg
     4741 gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac
     4801 gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga taataccatg
     4861 gccaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta
     4921 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgacccggac
     4981 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     5041 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     5101 agcgtcgaag cgggggcggt gttcgccgag atcggctcgc gcatggccaa gttgagcggt
     5161 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
     5221 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
     5281 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ctggggtgcc cgccttcctg
     5341 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     5401 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
     5461 gcggccgcaa tggatcctgt ttgaatgagg cttcagtact ttacagaatc gttgcctgca
     5521 catcttggaa acacttgctg ggattacttc ttcaggttaa cccaacagaa ggctaaagaa
     5581 ggtatattgc tgttgacagt gagcgnnnnn nnnnnnnnnn nnnnnnntag tgaagccaca
     5641 gatgtannnn nnnnnnnnnn nnnnnnnntg cctactgcct cggacttcaa ggggctactt
     5701 taggagcaat tatcttgttt actaaaactg aataccttgc tatctctttg atacattttt
     5761 acaaagctga attaaaatgg tataaattaa atcactttac gcgtggcgcg cctggaacaa
     5821 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     5881 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     5941 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     6001 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     6061 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     6121 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     6181 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc
     6241 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     6301 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     6361 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaattaattc
     6421 tgcagtcgag acctagaaaa acatggagca atcacaagta gcaatacagc agctaccaat
     6481 gctgattgtg cctggctaga agcacaagag gaggaggagg tgggtttttc cagtcacacc
     6541 tcaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
     6601 agaaaagagg ggactggaag ggctaattca ctcccaacga agacaagatc tgctttttgc
     6661 ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     6721 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     6781 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     6841 ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg
     6901 aatatcagag agtgagaggc cttgacattg tttaaactgg aatgtgtgtc agttagggtg
     6961 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     7021 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     7081 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     7141 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     7201 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     7261 aggcttttgc aaaaagctcc cgggagcttg tatatccatt acatgtgagc aaaaggccag
     7321 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     7381 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     7441 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     7501 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     7561 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     7621 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     7681 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     7741 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     7801 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     7861 agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag
     7921 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     7981 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     8041 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     8101 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     8161 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     8221 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     8281 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     8341 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     8401 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     8461 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     8521 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     8581 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     8641 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     8701 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     8761 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     8821 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     8881 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     8941 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     9001 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     9061 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
