LOCUS       V010346                 8955 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V010346
VERSION     V010346
KEYWORDS    pZIP-hEF1-alpha-ZsGreen-Puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8955)
  AUTHORS   transOMIC
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8955)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8955
                     /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             3415..4107
                     /label="ZsGreen1"
                     /note="Zoanthus green fluorescent protein"
     misc_feature    4124..4706
                     /label="IRES2"
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
     CDS             4705..5298
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     ncRNA           5322..5443
                     /label="5' miR-30a"
                     /note="sequence upstream of the 71-nt precursor of the
                     human miR-30a microRNA (Zeng et al., 2002)"
     misc_RNA        5444..5512
                     /label="shRNA"
                     /note="shRNA"
                     /note="short hairpin RNA"
     gap             5447..5468
                     /estimated_length=22
     gap             5488..5509
                     /estimated_length=22
     ncRNA           5513..5639
                     /label="3' miR-30a"
                     /note="sequence downstream of the 71-nt precursor of the
                     human miR-30a microRNA (Zeng et al., 2002)"
     misc_feature    5660..6248
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             6456..6689
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        6785..7114
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      complement(7203..7788)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7962..8819)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(8820..8924)
                     /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 gtccatggcc
     3421 cagtccaagc acggcctgac caaggagatg accatgaagt accgcatgga gggctgcgtg
     3481 gacggccaca agttcgtgat caccggcgag ggcatcggct accccttcaa gggcaagcag
     3541 gccatcaacc tgtgcgtggt ggagggcggc cccttgccct tcgccgagga catcttgtcc
     3601 gccgccttca tgtacggcaa ccgcgtgttc accgagtacc cccaggacat cgtcgactac
     3661 ttcaagaact cctgccccgc cggctacacc tgggaccgct ccttcctgtt cgaggacggc
     3721 gccgtgtgca tctgcaacgc cgacatcacc gtgagcgtgg aggagaactg catgtaccac
     3781 gagtccaagt tctacggcgt gaacttcccc gccgacggcc ccgtgatgaa gaagatgacc
     3841 gacaactggg agccctcctg cgagaagatc atccccgtgc ccaagcaggg catcttgaag
     3901 ggcgacgtga gcatgtacct gctgctgaag gacggtggcc gcttgcgctg ccagttcgac
     3961 accgtgtaca aggccaagtc cgtgccccgc aagatgcccg actggcactt catccagcac
     4021 aagctgaccc gcgaggaccg cagcgacgcc aagaaccaga agtggcacct gaccgagcac
     4081 gccatcgcct ccggctccgc cttgccctga gaattcaatt ccgcccctct ccctcccccc
     4141 cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt
     4201 tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct
     4261 tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga
     4321 atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga
     4381 ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac
     4441 gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag
     4501 ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc
     4561 agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg
     4621 tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt
     4681 gaaaaacacg ataataccat ggccaccgag tacaagccca cggtgcgcct cgccacccgc
     4741 gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     4801 cgccacaccg tcgacccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     4861 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     4921 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggctcg
     4981 cgcatggcca agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     5041 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     5101 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     5161 gctggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     5221 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     5281 acccgcaagc ccggtgcctg agcggccgca atggatcctg tttgaatgag gcttcagtac
     5341 tttacagaat cgttgcctgc acatcttgga aacacttgct gggattactt cttcaggtta
     5401 acccaacaga aggctaaaga aggtatattg ctgttgacag tgagcgnnnn nnnnnnnnnn
     5461 nnnnnnnnta gtgaagccac agatgtannn nnnnnnnnnn nnnnnnnnnt gcctactgcc
     5521 tcggacttca aggggctact ttaggagcaa ttatcttgtt tactaaaact gaataccttg
     5581 ctatctcttt gatacatttt tacaaagctg aattaaaatg gtataaatta aatcacttta
     5641 cgcgtggcgc gcctggaaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     5701 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     5761 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     5821 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     5881 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     5941 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     6001 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     6061 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     6121 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     6181 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     6241 ctccccgcct ggaattaatt ctgcagtcga gacctagaaa aacatggagc aatcacaagt
     6301 agcaatacag cagctaccaa tgctgattgt gcctggctag aagcacaaga ggaggaggag
     6361 gtgggttttt ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt
     6421 agatcttagc cactttttaa aagaaaagag gggactggaa gggctaattc actcccaacg
     6481 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg
     6541 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     6601 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     6661 cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta
     6721 tttataactt gcaaagaaat gaatatcaga gagtgagagg ccttgacatt gtttaaactg
     6781 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     6841 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     6901 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     6961 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     7021 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     7081 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     7141 tacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     7201 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     7261 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     7321 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     7381 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     7441 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     7501 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     7561 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     7621 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     7681 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     7741 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     7801 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     7861 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     7921 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     7981 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     8041 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     8101 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     8161 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     8221 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     8281 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     8341 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     8401 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     8461 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     8521 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     8581 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     8641 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     8701 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     8761 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     8821 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     8881 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     8941 gtgccacctg acgtc
//
