LOCUS       V010342                 9110 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V010342
VERSION     V010342
KEYWORDS    pZIP-mEF1-alpha-ZsGreen-Puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9110)
  AUTHORS   transOMIC
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 9110)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9110
                     /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             3570..4262
                     /label="ZsGreen1"
                     /note="Zoanthus green fluorescent protein"
     misc_feature    4279..4861
                     /label="IRES2"
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
     CDS             4860..5453
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     ncRNA           5477..5598
                     /label="5' miR-30a"
                     /note="sequence upstream of the 71-nt precursor of the
                     human miR-30a microRNA (Zeng et al., 2002)"
     misc_RNA        5599..5667
                     /label="shRNA"
                     /note="shRNA"
                     /note="short hairpin RNA"
     gap             5602..5623
                     /estimated_length=22
     gap             5643..5664
                     /estimated_length=22
     ncRNA           5668..5794
                     /label="3' miR-30a"
                     /note="sequence downstream of the 71-nt precursor of the
                     human miR-30a microRNA (Zeng et al., 2002)"
     misc_feature    5815..6403
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             6611..6844
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        6940..7269
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      complement(7358..7943)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(8117..8974)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(8975..9079)
                     /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 aaccggtcca tggcccagtc caagcacggc ctgaccaagg
     3601 agatgaccat gaagtaccgc atggagggct gcgtggacgg ccacaagttc gtgatcaccg
     3661 gcgagggcat cggctacccc ttcaagggca agcaggccat caacctgtgc gtggtggagg
     3721 gcggcccctt gcccttcgcc gaggacatct tgtccgccgc cttcatgtac ggcaaccgcg
     3781 tgttcaccga gtacccccag gacatcgtcg actacttcaa gaactcctgc cccgccggct
     3841 acacctggga ccgctccttc ctgttcgagg acggcgccgt gtgcatctgc aacgccgaca
     3901 tcaccgtgag cgtggaggag aactgcatgt accacgagtc caagttctac ggcgtgaact
     3961 tccccgccga cggccccgtg atgaagaaga tgaccgacaa ctgggagccc tcctgcgaga
     4021 agatcatccc cgtgcccaag cagggcatct tgaagggcga cgtgagcatg tacctgctgc
     4081 tgaaggacgg tggccgcttg cgctgccagt tcgacaccgt gtacaaggcc aagtccgtgc
     4141 cccgcaagat gcccgactgg cacttcatcc agcacaagct gacccgcgag gaccgcagcg
     4201 acgccaagaa ccagaagtgg cacctgaccg agcacgccat cgcctccggc tccgccttgc
     4261 cctgagaatt caattccgcc cctctccctc ccccccccct aacgttactg gccgaagccg
     4321 cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt
     4381 tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct
     4441 ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct
     4501 ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc
     4561 acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc
     4621 ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc
     4681 ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc
     4741 tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct
     4801 aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgataat accatggcca
     4861 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca
     4921 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc
     4981 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
     5041 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg
     5101 tcgaagcggg ggcggtgttc gccgagatcg gctcgcgcat ggccaagttg agcggttccc
     5161 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg
     5221 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg
     5281 ccgtcgtgct ccccggagtg gaggcggccg agcgcgctgg ggtgcccgcc ttcctggaga
     5341 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
     5401 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgagcgg
     5461 ccgcaatgga tcctgtttga atgaggcttc agtactttac agaatcgttg cctgcacatc
     5521 ttggaaacac ttgctgggat tacttcttca ggttaaccca acagaaggct aaagaaggta
     5581 tattgctgtt gacagtgagc gnnnnnnnnn nnnnnnnnnn nnntagtgaa gccacagatg
     5641 tannnnnnnn nnnnnnnnnn nnnntgccta ctgcctcgga cttcaagggg ctactttagg
     5701 agcaattatc ttgtttacta aaactgaata ccttgctatc tctttgatac atttttacaa
     5761 agctgaatta aaatggtata aattaaatca ctttacgcgt ggcgcgcctg gaacaatcaa
     5821 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     5881 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     5941 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     6001 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     6061 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     6121 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     6181 actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt
     6241 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     6301 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     6361 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaat taattctgca
     6421 gtcgagacct agaaaaacat ggagcaatca caagtagcaa tacagcagct accaatgctg
     6481 attgtgcctg gctagaagca caagaggagg aggaggtggg tttttccagt cacacctcag
     6541 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     6601 aagaggggac tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt
     6661 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     6721 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     6781 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     6841 agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata
     6901 tcagagagtg agaggccttg acattgttta aactggaatg tgtgtcagtt agggtgtgga
     6961 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     7021 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     7081 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     7141 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     7201 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     7261 ttttgcaaaa agctcccggg agcttgtata tccattacat gtgagcaaaa ggccagcaaa
     7321 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     7381 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     7441 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     7501 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     7561 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     7621 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     7681 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     7741 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     7801 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     7861 cttgatccgg caaacaaacc accgctggta gcggtttttt tgtttgcaag cagcagatta
     7921 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     7981 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     8041 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     8101 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     8161 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     8221 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     8281 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     8341 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     8401 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     8461 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     8521 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     8581 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     8641 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     8701 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     8761 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     8821 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     8881 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     8941 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     9001 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     9061 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
