LOCUS       V010477                 7695 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V010477
VERSION     V010477
KEYWORDS    pLenti-C-Myc-DDK-IRES-tRFP.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7695)
  AUTHORS   OriGene
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7695)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
            Inserts are typically cloned between the SgfI (AsiSI) and MluI
            sites.
FEATURES             Location/Qualifiers
     source          1..7695
                     /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"
     LTR             834..1014
                     /label="5' LTR (truncated)"
                     /note="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     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"
     enhancer        2643..3022
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3024..3223
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        3252..3270
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    3279..3387
                     /label="MCS"
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             3385..3414
                     /label="Myc"
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
     CDS             3433..3456
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     misc_feature    3490..4067
                     /label="IRES2"
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
     CDS             4065..4757
                     /label="TurboRFP"
                     /note="red fluorescent protein from Entacmaea quadricolor"
     protein_bind    complement(4790..4823)
                     /label="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4879..5467
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     primer_bind     complement(5470..5486)
                     /label="KS primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     LTR             5996..6176
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6238..6826)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6906..7562)
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(7563..7667)
                     /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 gctctagaca tgtccaatat gaccgccatg
     2641 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
     2701 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     2761 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     2821 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     2881 tcaagtgtat catatgccaa gtccgccccc tattgacgtc aatgacggta aatggcccgc
     2941 ctggcattat gcccagtaca tgaccttacg ggactttcct acttggcagt acatctacgt
     3001 attagtcatc gctattacca tggtgatgcg gttttggcag tacaccaatg ggcgtggata
     3061 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     3121 ttggcaccaa aatcaacggg actttccaaa atgtcgtaat aaccccgccc cgttgacgca
     3181 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg
     3241 tcagaatttt gtaatacgac tcactatagg gcggccggga attcgtcgac tggatccggt
     3301 accgaggaga tctgccgccg cgatcgccgg cgcgccagat ctcaagctta actagctagc
     3361 ggaccgacgc gtacgcggcc gctcgagcag aaactcatct cagaagagga tctggcagca
     3421 aatgatatcc tggattacaa ggatgacgac gataaggttt aaacctaggc gtagcggccg
     3481 caaattccgc ccctctccct cccccccccc taacgttact ggccgaagcc gcttggaata
     3541 aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt
     3601 gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct
     3661 cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc
     3721 ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga
     3781 caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc
     3841 ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt
     3901 attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg
     3961 gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc
     4021 gaaccacggg gacgtggttt tcctttgaaa aacacgatga tcatatgagc gagctgatca
     4081 aggagaacat gcacatgaag ctgtacatgg agggcaccgt gaacaaccac cacttcaagt
     4141 gcacatccga gggcgaaggc aagccctacg agggcaccca gaccatgaag atcaaggtgg
     4201 tcgagggcgg ccctctcccc ttcgccttcg acatcctggc taccagcttc atgtacggca
     4261 gcaaagcctt catcaaccac acccagggca tccccgactt ctttaagcag tccttccctg
     4321 agggcttcac atgggagaga atcaccacat acgaagacgg gggcgtgctg accgctaccc
     4381 aggacaccag cttccagaac ggctgcatca tctacaacgt caagatcaac ggggtgaact
     4441 tcccatccaa cggccctgtg atgcagaaga aaacacgcgg ctgggaggcc aacaccgaga
     4501 tgctgtaccc cgctgacggc ggcctgagag gccacagcca gatggccctg aagctcgtgg
     4561 gcgggggcta cctgcactgc tccttcaaga ccacatacag atccaagaaa cccgctaaga
     4621 acctcaagat gcccggcttc cacttcgtgg accacagact ggaaagaatc aaggaggccg
     4681 acaaagagac ctacgtcgag cagcacgaga tggctgtggc caagtactgc gacctcccta
     4741 gcaaactggg gcacagatga tgtacaagta ggattcgtcg agggacctaa taacttcgta
     4801 tagcatacat tatacgaagt tatacatgtt taagggttcc ggttccacta ggtacaattc
     4861 gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     4921 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     4981 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     5041 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     5101 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     5161 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     5221 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     5281 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     5341 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     5401 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     5461 tccccgcatc gataccgtcg acctcgatcg agacctagaa aaacatggag caatcacaag
     5521 tagcaataca gcagctacca atgctgattg tgcctggcta gaagcacaag aggaggagga
     5581 ggtgggtttt ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt
     5641 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
     5701 aagacaagat atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattggca
     5761 gaactacaca ccagggccag ggatcagata tccactgacc tttggatggt gctacaagct
     5821 agtaccagtt gagcaagaga aggtagaaga agccaatgaa ggagagaaca cccgcttgtt
     5881 acaccctgtg agcctgcatg ggatggatga cccggagaga gaagtattag agtggaggtt
     5941 tgacagccgc ctagcatttc atcacatggc ccgagagctg catccggact gtactgggtc
     6001 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     6061 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     6121 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagcat
     6181 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6241 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6301 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6361 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6421 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6481 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6541 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6601 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6661 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6721 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6781 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6841 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6901 gattacgccc cgccctgcca ctcatcgcag tactgttgta attcattaag cattctgccg
     6961 acatggaagc catcacaaac ggcatgatga acctgaatcg ccagcggcat cagcaccttg
     7021 tcgccttgcg tataatattt gcccatggtg aaaacggggg cgaagaagtt gtccatattg
     7081 gccacgttta aatcaaaact ggtgaaactc acccagggat tggctgagac gaaaaacata
     7141 ttctcaataa accctttagg gaaataggcc aggttttcac cgtaacacgc cacatcttgc
     7201 gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt cactccagag cgatgaaaac
     7261 gtttcagttt gctcatggaa aacggtgtaa caagggtgaa cactatccca tatcaccagc
     7321 tcaccgtctt tcattgccat acggaactcc ggatgagcat tcatcaggcg ggcaagaatg
     7381 tgaataaagg ccggataaaa cttgtgctta tttttcttta cggtctttaa aaaggccgta
     7441 atatccagct gaacggtctg gttataggta cattgagcaa ctgactgaaa tgcctcaaaa
     7501 tgttctttac gatgccattg ggatatatca acggtggtat atccagtgat ttttttctcc
     7561 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7621 tacatatttg aatgtattta gaaaaataaa caaatagggg tcccgcgcac atttccccga
     7681 aaagtgccac ctgac
//
