LOCUS       pLVX-Puro-Cnr1(rat)     9505 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040784
VERSION     V040784
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             1..634
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     misc_feature    681..806
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1303..1536
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1721..1765
                     /codon_start=1
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /product="antigenic peptide corresponding to amino acids
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
                     et al., 2013)"
                     /transl_table=1
                     /translation="KNEQELLELDKWASL"
                     /label="gp41 peptide"
     misc_feature    2028..2143
                     /note="central polypurine tract and central termination
                     sequence of HIV-1 (lacking the first T)"
                     /label="cPPT/CTS"
     enhancer        2201..2504
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2505..2708
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2843..4261
                     /label="Cnr1(NM_012784)"
                     /note="Cnr1(NM_012784)"
                     /gene="Cnr1"
     promoter        4293..4792
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4813..5412
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     misc_feature    5426..6014
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6221..6854
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6983..6999)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7007..7023
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(7031..7061)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7076..7097
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7385..7973)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8144..9004)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(9005..9109)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9157..9291
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa
     2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     2761 tccatagaag acaccgactc tactagagga tcgctagcgc taccggactc agatctcgag
     2821 ctcaagcttc gaattcgcca ccatgaagtc gatcctagat ggccttgcag acaccacctt
     2881 ccgtaccatc accacagacc tcctctacgt gggctcgaat gacattcagt atgaagatat
     2941 caaaggagac atggcatcca aattaggata cttcccacag aaattccctc taacttcctt
     3001 caggggtagt cccttccaag aaaagatgac cgcaggagac aactccccgt tggtcccagc
     3061 aggagacaca acaaacatta cagagttcta taacaagtct ctctcgtcgt tcaaggagaa
     3121 tgaggagaac atccagtgtg gggagaactt tatggacatg gagtgcttta tgattctgaa
     3181 tcccagccag cagctggcca tcgctgtact gtccctcaca ctgggcacct tcacggttct
     3241 ggagaaccta ctggtgctgt gtgtcatcct gcactcccgc agtctccgat gcaggccttc
     3301 ctaccacttc atcggcagcc tggcagtggc cgacctcctg ggaagtgtca tttttgtgta
     3361 cagctttgtt gacttccatg tattccaccg taaagacagc cccaatgtgt ttctgttcaa
     3421 actgggtggg gttacagcct ccttcacagc ttctgtgggc agcctgttcc tcacagccat
     3481 cgacaggtac atatccattc acaggcctct ggcctataag aggatcgtca ccaggcccaa
     3541 ggccgttgtg gccttttgcc tgatgtggac tatcgcaata gtaatcgctg tgttgcctct
     3601 cctgggctgg aactgcaaga agctgcaatc tgtttgctcg gacattttcc cactcattga
     3661 cgagacctac ctgatgttct ggattggggt gaccagtgtg ctgctgctgt tcattgtgta
     3721 cgcgtacatg tacattctct ggaaggctca cagccatgcg gtccgcatga ttcagcgtgg
     3781 gacccagaag agcatcatca tccacacgtc agaagacggc aaggtgcagg tgacccggcc
     3841 tgaccaagcc cgcatggaca ttaggctggc caaaaccctg gttctgatcc tggtggtgtt
     3901 gatcatctgc tggggccctc tgcttgcgat catggtgtat gacgtcttcg ggaagatgaa
     3961 caagcttatc aagacggtgt ttgccttctg cagtatgctc tgcctgctga actccaccgt
     4021 gaaccccatc atctatgctc tgaggagcaa ggacctgaga catgctttcc gaagcatgtt
     4081 cccttcgtgc gaaggcaccg cacagcctct agacaacagc atgggggact cagactgcct
     4141 gcacaagcac gccaacaaca cagccagcat gcacagggcc gcggagagct gcatcaagag
     4201 caccgttaag atcgcgaagg tgaccatgtc tgtgtccaca gacacgtccg ccgaggctct
     4261 gtgaggatcc cgcgactcta gataattcta ccgggtaggg gaggcgcttt tcccaaggca
     4321 gtctggagca tgcgctttag cagccccgct gggcacttgg cgctacacaa gtggcctctg
     4381 gcctcgcaca cattccacat ccaccggtag gcgccaaccg gctccgttct ttggtggccc
     4441 cttcgcgcca ccttctactc ctcccctagt caggaagttc ccccccgccc cgcagctcgc
     4501 gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact agtctcgtgc agatggacag
     4561 caccgctgag caatggaagc gggtaggcct ttggggcagc ggccaatagc agctttgctc
     4621 cttcgctttc tgggctcaga ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg
     4681 gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc ccggcattct gcacgcttca
     4741 aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct ccgggccttt cgacctgcag
     4801 cccaagctta ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     4861 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     4921 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     4981 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     5041 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     5101 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     5161 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     5221 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     5281 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     5341 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     5401 cccggtgcct gaccgcgtct ggaacaatca acctctggat tacaaaattt gtgaaagatt
     5461 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     5521 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     5581 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     5641 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     5701 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     5761 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     5821 gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     5881 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     5941 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     6001 ggccgcctcc ccgcctggaa ttaattctgc agtcgagacc tagaaaaaca tggagcaatc
     6061 acaagtagca atacagcagc taccaatgct gattgtgcct ggctagaagc acaagaggag
     6121 gaggaggtgg gttttccagt cacacctcag gtacctttaa gaccaatgac ttacaaggca
     6181 gctgtagatc ttagccactt tttaaaagaa aagaggggac tggaagggct aattcactcc
     6241 caacgaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta cttccctgat
     6301 tagcagaact acacaccagg gccaggggtc agatatccac tgacctttgg atggtgctac
     6361 aagctagtac cagttgagcc agataaggta gaagaggcca ataaaggaga gaacaccagc
     6421 ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt gttagagtgg
     6481 aggtttgaca gccgcctagc atttcatcac gtggcccgag agctgcatcc ggagtacttc
     6541 aagaactgct gatatcgagc ttgctacaag ggactttccg ctggggactt tccagggagg
     6601 cgtggcctgg gcgggactgg ggagtggcga gccctcagat cctgcatata agcagctgct
     6661 ttttgcctgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     6721 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     6781 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     6841 gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa
     6901 gaaatgaata tcagagagtg agaggccttg acattgctag cgttttaccg tcgacctcta
     6961 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     7021 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     7081 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     7141 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     7201 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     7261 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     7321 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     7381 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     7441 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     7501 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     7561 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     7621 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     7681 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     7741 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     7801 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     7861 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     7921 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     7981 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     8041 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     8101 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     8161 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     8221 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     8281 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     8341 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     8401 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     8461 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     8521 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     8581 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     8641 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     8701 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     8761 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     8821 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     8881 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     8941 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     9001 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     9061 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     9121 gaaaagtgcc acctgacgtc gacggatcgg gagatcaact tgtttattgc agcttataat
     9181 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     9241 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat caactggata
     9301 actcaagcta accaaaatca tcccaaactt cccaccccat accctattac cactgccaat
     9361 tacctgtggt ttcatttact ctaaacctgt gattcctctg aattattttc attttaaaga
     9421 aattgtattt gttaaatatg tactacaaac ttagtagttt ttaaagaaat tgtatttgtt
     9481 aaatatgtac tacaaactta gtagt
//
