LOCUS       V010469                 7040 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V010469
VERSION     V010469
KEYWORDS    pLenti-N-tRFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7040)
  AUTHORS   OriGene
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7040)
  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..7040
                     /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="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"
     CDS             3320..4012
                     /label="TurboRFP"
                     /note="red fluorescent protein from Entacmaea quadricolor"
     misc_feature    4013..4091
                     /label="MCS"
                     /note="MCS"
                     /note="multiple cloning site"
     protein_bind    complement(4135..4168)
                     /label="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4224..4812
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     primer_bind     complement(4815..4831)
                     /label="KS primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     LTR             5341..5521
                     /label="3' LTR (truncated)"
                     /note="3' LTR (truncated)"
                     /note="truncated 3' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(5583..6171)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6251..6907)
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(6908..7012)
                     /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 tctgccgcca tgagcgagct gatcaaggag aacatgcaca tgaagctgta
     3361 catggagggc accgtgaaca accaccactt caagtgcaca tccgagggcg aaggcaagcc
     3421 ctacgagggc acccagacca tgaagatcaa ggtggtcgag ggcggccctc tccccttcgc
     3481 cttcgacatc ctggctacca gcttcatgta cggcagcaaa gccttcatca accacaccca
     3541 gggcatcccc gacttcttta agcagtcctt ccctgagggc ttcacatggg agagaatcac
     3601 cacatacgaa gacgggggcg tgctgaccgc tacccaggac accagcttcc agaacggctg
     3661 catcatctac aacgtcaaga tcaacggggt gaacttccca tccaacggcc ctgtgatgca
     3721 gaagaaaaca cgcggctggg aggccaacac cgagatgctg taccccgctg acggcggcct
     3781 gagaggccac agccagatgg ccctgaagct cgtgggcggg ggctacctgc actgctcctt
     3841 caagaccaca tacagatcca agaaacccgc taagaacctc aagatgcccg gcttccactt
     3901 cgtggaccac agactggaaa gaatcaagga ggccgacaaa gagacctacg tcgagcagca
     3961 cgagatggct gtggccaagt actgcgacct ccctagcaaa ctggggcaca gagcgatcgc
     4021 cggcgcgcca gatctcaagc ttaactagtt agcggaccga cgcgttaagc ggccgcactc
     4081 gaggtttaaa cggccggccg cggtctgtac aagtaggatt cgtcgaggga cctaataact
     4141 tcgtatagca tacattatac gaagttatac atgtttaagg gttccggttc cactaggtac
     4201 aattcgatat caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga
     4261 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     4321 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     4381 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     4441 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     4501 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     4561 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     4621 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     4681 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     4741 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     4801 ccgcctcccc gcatcgatac cgtcgacctc gatcgagacc tagaaaaaca tggagcaatc
     4861 acaagtagca atacagcagc taccaatgct gattgtgcct ggctagaagc acaagaggag
     4921 gaggaggtgg gttttccagt cacacctcag gtacctttaa gaccaatgac ttacaaggca
     4981 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc
     5041 caacgaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta cttccctgat
     5101 tggcagaact acacaccagg gccagggatc agatatccac tgacctttgg atggtgctac
     5161 aagctagtac cagttgagca agagaaggta gaagaagcca atgaaggaga gaacacccgc
     5221 ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt attagagtgg
     5281 aggtttgaca gccgcctagc atttcatcac atggcccgag agctgcatcc ggactgtact
     5341 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     5401 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     5461 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     5521 agcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5581 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5641 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5701 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5761 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5821 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5881 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5941 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6001 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     6061 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6121 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6181 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6241 gtcatgatta cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc
     6301 tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca
     6361 ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca
     6421 tattggccac gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa
     6481 acatattctc aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat
     6541 cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg
     6601 aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca
     6661 ccagctcacc gtctttcatt gccatacgga actccggatg agcattcatc aggcgggcaa
     6721 gaatgtgaat aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg
     6781 ccgtaatatc cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct
     6841 caaaatgttc tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt
     6901 tctccatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6961 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggtcccg cgcacatttc
     7021 cccgaaaagt gccacctgac
//
