LOCUS       V010345                 8308 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V010345
VERSION     V010345
KEYWORDS    pZIP-mCMV-RFP-Puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8308)
  AUTHORS   transOMIC
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8308)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8308
                     /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..2755
                     /label="mCMV promoter"
                     /note="mouse cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             2768..3460
                     /label="TurboRFP"
                     /note="red fluorescent protein from Entacmaea quadricolor"
     misc_feature    3477..4059
                     /label="IRES2"
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
     CDS             4058..4651
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     ncRNA           4675..4796
                     /label="5' miR-30a"
                     /note="sequence upstream of the 71-nt precursor of the
                     human miR-30a microRNA (Zeng et al., 2002)"
     misc_RNA        4797..4865
                     /label="shRNA"
                     /note="shRNA"
                     /note="short hairpin RNA"
     gap             4800..4821
                     /estimated_length=22
     gap             4841..4862
                     /estimated_length=22
     ncRNA           4866..4992
                     /label="3' miR-30a"
                     /note="sequence downstream of the 71-nt precursor of the
                     human miR-30a microRNA (Zeng et al., 2002)"
     misc_feature    5013..5601
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             5809..6042
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        6138..6467
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      complement(6556..7141)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7315..8172)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(8173..8277)
                     /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 ttatcgattg
     2221 gcacgtatac tgagtcatta gggactttcc aatgggtttt gcccagtaca taaggtcaat
     2281 aggggtgaat caacaggaaa gtcccattgg agccaagtac actgagtcaa tagggacttt
     2341 ccattgggtt ttgcccagta caaaaggtca atagggggtg agtcaatggg tttttcccat
     2401 tattggcacg tacataaggt caataggggt gagtcattgg gtttttccag ccaatttaat
     2461 taaaacgcca tgtactttcc caccattgac gtcaatgggc tattgaaact aatgcaacgt
     2521 gacctttaaa cggtactttc ccatagctga ttaatgggaa agtaccgttc tcagagccaa
     2581 tacacgtcaa tgggaagtga aagggcagcc aaaacgtaac accgccccgg ttttcccctg
     2641 gaaattccat attggcacgc attctattgg ctgagctgcg ttctacgtgg gtataagagg
     2701 cgcgaccagc gtcggtaccg tcgcagtctt cggtctgacc accgtagaac gcagaaccgg
     2761 tgccaccatg agcgagctga tcaaggagaa catgcacatg aagctgtaca tggagggcac
     2821 cgtgaacaac caccacttca agtgcacatc cgagggcgaa ggcaagccct acgagggcac
     2881 ccagaccatg aagatcaagg tggtcgaggg cggccctctc cccttcgcct tcgacatcct
     2941 ggctaccagc ttcatgtacg gcagcaaagc cttcatcaac cacacccagg gcatccccga
     3001 cttctttaag cagtccttcc ctgagggctt cacatgggag agaatcacca catacgaaga
     3061 cgggggcgtg ctgaccgcta cccaggacac cagcttccag aacggctgca tcatctacaa
     3121 cgtcaagatc aacggggtga acttcccatc caacggccct gtgatgcaga agaaaacacg
     3181 cggctgggag gccaacaccg agatgctgta ccccgctgac ggcggcctga gaggccacag
     3241 ccagatggcc ctgaagctcg tgggcggggg ctacctgcac tgctccttca agaccacata
     3301 cagatccaag aaacccgcta agaacctcaa gatgcccggc ttccacttcg tggaccacag
     3361 actggaaaga atcaaggagg ccgacaaaga gacctacgtc gagcagcacg agatggctgt
     3421 ggccaagtac tgcgacctcc ctagcaaact ggggcacaga taagaattca attccgcccc
     3481 tctccctccc ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg
     3541 tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa
     3601 cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg
     3661 caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca
     3721 acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc
     3781 ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt
     3841 gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg
     3901 ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca
     3961 tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac
     4021 gtggttttcc tttgaaaaac acgataatac catggccacc gagtacaagc ccacggtgcg
     4081 cctcgccacc cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg cgttcgccga
     4141 ctaccccgcc acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg tcaccgagct
     4201 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     4261 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     4321 cgagatcggc tcgcgcatgg ccaagttgag cggttcccgg ctggccgcgc agcaacagat
     4381 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     4441 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     4501 ggcggccgag cgcgctgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     4561 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     4621 cacctggtgc atgacccgca agcccggtgc ctgagcggcc gcaatggatc ctgtttgaat
     4681 gaggcttcag tactttacag aatcgttgcc tgcacatctt ggaaacactt gctgggatta
     4741 cttcttcagg ttaacccaac agaaggctaa agaaggtata ttgctgttga cagtgagcgn
     4801 nnnnnnnnnn nnnnnnnnnn ntagtgaagc cacagatgta nnnnnnnnnn nnnnnnnnnn
     4861 nntgcctact gcctcggact tcaaggggct actttaggag caattatctt gtttactaaa
     4921 actgaatacc ttgctatctc tttgatacat ttttacaaag ctgaattaaa atggtataaa
     4981 ttaaatcact ttacgcgtgg cgcgcctgga acaatcaacc tctggattac aaaatttgtg
     5041 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     5101 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     5161 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     5221 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     5281 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     5341 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     5401 cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg
     5461 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     5521 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     5581 ccctttgggc cgcctccccg cctggaatta attctgcagt cgagacctag aaaaacatgg
     5641 agcaatcaca agtagcaata cagcagctac caatgctgat tgtgcctggc tagaagcaca
     5701 agaggaggag gaggtgggtt tttccagtca cacctcaggt acctttaaga ccaatgactt
     5761 acaaggcagc tgtagatctt agccactttt taaaagaaaa gaggggactg gaagggctaa
     5821 ttcactccca acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc
     5881 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     5941 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     6001 gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc
     6061 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggccttgac
     6121 attgtttaaa ctggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     6181 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     6241 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     6301 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     6361 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     6421 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     6481 cttgtatatc cattacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6541 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6601 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6661 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6721 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     6781 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6841 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6901 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6961 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     7021 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     7081 cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     7141 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7201 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7261 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7321 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7381 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7441 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7501 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7561 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7621 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7681 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7741 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7801 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7861 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7921 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7981 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     8041 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     8101 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     8161 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     8221 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     8281 atttccccga aaagtgccac ctgacgtc
//
