LOCUS       V014045                 9353 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014045
VERSION     V014045
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9353)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9353
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        237..616
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..819
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     misc_feature    1061..1186
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1683..1916
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             2101..2145
                     /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             2294..2335
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    2443..2560
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2624..3802
                     /label="EF-1-alpha promoter"
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     CDS             3919..3942
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             3952..3969
                     /label="6xHis"
                     /note="6xHis affinity tag"
     promoter        4413..4612
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        4641..4659
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4701..5366
                     /label="CopGFP"
                     /note="green fluorescent protein 2 from Pontellina plumata,
                     also known as ppluGFP2 (Shagin et al., 2004)"
     CDS             5444..5500
                     /label="P2A"
                     /note="2A peptide from porcine teschovirus-1 polyprotein"
     CDS             5501..6097
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     protein_bind    complement(6153..6186)
                     /label="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    6242..6830
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     primer_bind     complement(6833..6849)
                     /label="KS primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     LTR             7359..7539
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(7601..8189)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(8363..9220)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(9221..9325)
                     /label="AmpR promoter"
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc
      841 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      901 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      961 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
     1021 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg
     1081 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat
     1141 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg
     1201 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa
     1261 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct
     1321 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac
     1381 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca
     1441 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa
     1501 aagtaagacc accgcacagc aagcggccgg ccgctgatct tcagacctgg aggaggagat
     1561 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1621 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1681 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1741 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1801 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1861 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1921 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1981 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     2041 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     2101 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     2161 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     2221 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2281 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2341 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2401 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2461 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2521 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2581 cagagatcca gtttggttag taccgggccc gctctagacg tgaggctccg gtgcccgtca
     2641 gtgggcagag cgcacatcgc ccacagtccc cgagaagttg tggggagggg tcggcaattg
     2701 aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct
     2761 ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt
     2821 tctttttcgc aacgggtttg ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg
     2881 gcctggcctc tttacgggtt atggcccttg cgtgccttga attacttcca cctggctgca
     2941 gtacgtgatt cttgatcccg agcttcgggt tggaagtggg tgggagagtt cgaggccttg
     3001 cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc tggcctgggc gctggggccg
     3061 ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct gctttcgata agtctctagc
     3121 catttaaaat ttttgatgac ctgctgcgac gctttttttc tggcaagata gtcttgtaaa
     3181 tgcgggccaa gatctgcaca ctggtatttc ggtttttggg gccgcgggcg gcgacggggc
     3241 ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg cgagcgcggc caccgagaat
     3301 cggacggggg tagtctcaag ctggccggcc tgctctggtg cctggcctcg cgccgccgtg
     3361 tatcgccccg ccctgggcgg caaggctggc ccggtcggca ccagttgcgt gagcggaaag
     3421 atggccgctt cccggccctg ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga
     3481 gcgggcgggt gagtcaccca cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc
     3541 atgtgactcc acggagtacc gggcgccgtc caggcacctc gattagttct cgagcttttg
     3601 gagtacgtcg tctttaggtt ggggggaggg gttttatgcg atggagtttc cccacactga
     3661 gtgggtggag actgaagtta ggccagcttg gcacttgatg taattctcct tggaatttgc
     3721 cctttttgag tttggatctt ggttcattct caagcctcag acagtggttc aaagtttttt
     3781 tcttccattt caggtgtcgt gaggaatcct ctaggtcgac ctggatccgg taccgaggag
     3841 atctgccgcc gcgatcgccg gcgcgccaga tctcaagctt aactagctag cggaccgacg
     3901 cgtacgcggc cgctcgagga ttataaggat gacgacgata aattcgtcga gcaccaccac
     3961 caccaccact aataaggttt atccgatcca ccggaatgca ttctagacat gtccaatatg
     4021 accgccatgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
     4081 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     4141 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     4201 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     4261 ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca atgacggtaa
     4321 atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta cttggcagta
     4381 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acaccaatgg
     4441 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     4501 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata accccgcccc
     4561 gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcgttt
     4621 agtgaaccgt cagaattttg taatacgact cactataggg cggccgggaa ttcgccacca
     4681 tggagagcga cgagagcggc ctgcccgcca tggagatcga gtgccgcatc accggcaccc
     4741 tgaacggcgt ggagttcgag ctggtgggcg gcggagaggg cacccccaag cagggccgca
     4801 tgaccaacaa gatgaagagc accaaaggcg ccctgacctt cagcccctac ctgctgagcc
     4861 acgtgatggg ctacggcttc taccacttcg gcacctaccc cagcggctac gagaacccct
     4921 tcctgcacgc catcaacaac ggcggctaca ccaacacccg catcgagaag tacgaggacg
     4981 gcggcgtgct gcacgtgagc ttcagctacc gctacgaggc cggccgcgtg atcggcgact
     5041 tcaaggtggt gggcaccggc ttccccgagg acagcgtgat cttcaccgac aagatcatcc
     5101 gcagcaacgc caccgtggag cacctgcacc ccatgggcga taacgtgctg gtgggcagct
     5161 tcgcccgcac cttcagcctg cgcgacggcg gctactacag cttcgtggtg gacagccaca
     5221 tgcacttcaa gagcgccatc caccccagca tcctgcagaa cgggggcccc atgttcgcct
     5281 tccgccgcgt ggaggagctg cacagcaaca ccgagctggg catcgtggag taccagcacg
     5341 ccttcaagac ccccatcgcc ttcgccagat cccgcgctca gtcgtccaat tctgccgtgg
     5401 acggcaccgc cggacccggc tccaccggat ctcgcgaagc ggagctacta acttcagcct
     5461 gctgaagcag gctggagacg tggaggagaa ccctggacct atgaccgagt acaagcccac
     5521 ggtgcgcctc gccacccgcg acgacgtccc ccgggcagta cgcaccctcg ccgccgcgtt
     5581 cgccgactac cccgccacgc gccacaccgt cgatccagac cgccacatcg agcgggtcac
     5641 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     5701 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     5761 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     5821 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     5881 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     5941 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     6001 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     6061 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga gtttaaacgg gccggccgcg
     6121 gtctgtacaa gtaggattcg tcgagggacc taataacttc gtatagcata cattatacga
     6181 agttatacat gtttaagggt tccggttcca ctaggtacaa ttcgatatca agcttatcga
     6241 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     6301 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     6361 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     6421 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     6481 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     6541 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     6601 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     6661 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     6721 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     6781 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg
     6841 tcgacctcga tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta
     6901 ccaatgctga ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca
     6961 cacctcaggt acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
     7021 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg
     7081 atctgtggat ctaccacaca caaggctact tccctgattg gcagaactac acaccagggc
     7141 cagggatcag atatccactg acctttggat ggtgctacaa gctagtacca gttgagcaag
     7201 agaaggtaga agaagccaat gaaggagaga acacccgctt gttacaccct gtgagcctgc
     7261 atgggatgga tgacccggag agagaagtat tagagtggag gtttgacagc cgcctagcat
     7321 ttcatcacat ggcccgagag ctgcatccgg actgtactgg gtctctctgg ttagaccaga
     7381 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     7441 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     7501 ccctcagacc cttttagtca gtgtggaaaa tctctagcag catgtgagca aaaggccagc
     7561 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     7621 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     7681 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     7741 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     7801 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     7861 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     7921 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     7981 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     8041 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     8101 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     8161 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     8221 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     8281 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     8341 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     8401 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     8461 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     8521 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     8581 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     8641 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     8701 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     8761 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     8821 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     8881 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     8941 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     9001 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     9061 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     9121 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     9181 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     9241 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     9301 aaaataaaca aataggggtc ccgcgcacat ttccccgaaa agtgccacct gac
//
