LOCUS       V018373                10395 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018373
VERSION     V018373
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"
     regulatory      2821..2830
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             2830..2853
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             2854..5163
                     /label="APP(NM_000484)"
                     /note="APP(NM_000484)"
                     /gene="APP"
     promoter        5183..5682
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5703..6302
                     /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    6316..6904
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             7111..7744
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7873..7889)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7897..7913
                     /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(7921..7951)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7966..7987
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8275..8863)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9034..9894)
                     /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(9895..9999)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    10047..10181
                     /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 gccaccatgg actacaagga cgacgatgat aagatgctgc ccggtttggc actgctcctg
     2881 ctggccgcct ggacggctcg ggcgctggag gtacccactg atggtaatgc tggcctgctg
     2941 gctgaacccc agattgccat gttctgtggc agactgaaca tgcacatgaa tgtccagaat
     3001 gggaagtggg attcagatcc atcagggacc aaaacctgca ttgataccaa ggaaggcatc
     3061 ctgcagtatt gccaagaagt ctaccctgaa ctgcagatca ccaatgtggt agaagccaac
     3121 caaccagtga ccatccagaa ctggtgcaag cggggccgca agcagtgcaa gacccatccc
     3181 cactttgtga ttccctaccg ctgcttagtt ggtgagtttg taagtgatgc ccttctcgtt
     3241 cctgacaagt gcaaattctt acaccaggag aggatggatg tttgcgaaac tcatcttcac
     3301 tggcacaccg tcgccaaaga gacatgcagt gagaagagta ccaacttgca tgactacggc
     3361 atgttgctgc cctgcggaat tgacaagttc cgaggggtag agtttgtgtg ttgcccactg
     3421 gctgaagaaa gtgacaatgt ggattctgct gatgcggagg aggatgactc ggatgtctgg
     3481 tggggcggag cagacacaga ctatgcagat gggagtgaag acaaagtagt agaagtagca
     3541 gaggaggaag aagtggctga ggtggaagaa gaagaagccg atgatgacga ggacgatgag
     3601 gatggtgatg aggtagagga agaggctgag gaaccctacg aagaagccac agagagaacc
     3661 accagcattg ccaccaccac caccaccacc acagagtctg tggaagaggt ggttcgagag
     3721 gtgtgctctg aacaagccga gacggggccg tgccgagcaa tgatctcccg ctggtacttt
     3781 gatgtgactg aagggaagtg tgccccattc ttttacggcg gatgtggcgg caaccggaac
     3841 aactttgaca cagaagagta ctgcatggcc gtgtgtggca gcgccatgtc ccaaagttta
     3901 ctcaagacta cccaggaacc tcttgcccga gatcctgtta aacttcctac aacagcagcc
     3961 agtacccctg atgccgttga caagtatctc gagacacctg gggatgagaa tgaacatgcc
     4021 catttccaga aagccaaaga gaggcttgag gccaagcacc gagagagaat gtcccaggtc
     4081 atgagagaat gggaagaggc agaacgtcaa gcaaagaact tgcctaaagc tgataagaag
     4141 gcagttatcc agcatttcca ggagaaagtg gaatctttgg aacaggaagc agccaacgag
     4201 agacagcagc tggtggagac acacatggcc agagtggaag ccatgctcaa tgaccgccgc
     4261 cgcctggccc tggagaacta catcaccgct ctgcaggctg ttcctcctcg gcctcgtcac
     4321 gtgttcaata tgctaaagaa gtatgtccgc gcagaacaga aggacagaca gcacacccta
     4381 aagcatttcg agcatgtgcg catggtggat cccaagaaag ccgctcagat ccggtcccag
     4441 gttatgacac acctccgtgt gatttatgag cgcatgaatc agtctctctc cctgctctac
     4501 aacgtgcctg cagtggccga ggagattcag gatgaagttg atgagctgct tcagaaagag
     4561 caaaactatt cagatgacgt cttggccaac atgattagtg aaccaaggat cagttacgga
     4621 aacgatgctc tcatgccatc tttgaccgaa acgaaaacca ccgtggagct ccttcccgtg
     4681 aatggagagt tcagcctgga cgatctccag ccgtggcatt cttttggggc tgactctgtg
     4741 ccagccaaca cagaaaacga agttgagcct gttgatgccc gccctgctgc cgaccgagga
     4801 ctgaccactc gaccaggttc tgggttgaca aatatcaaga cggaggagat ctctgaagtg
     4861 aagatggatg cagaattccg acatgactca ggatatgaag ttcatcatca aaaattggtg
     4921 ttctttgcag aagatgtggg ttcaaacaaa ggtgcaatca ttggactcat ggtgggcggt
     4981 gttgtcatag cgacagtgat cgtcatcacc ttggtgatgc tgaagaagaa acagtacaca
     5041 tccattcatc atggtgtggt ggaggttgac gccgctgtca ccccagagga gcgccacctg
     5101 tccaagatgc agcagaacgg ctacgaaaat ccaacctaca agttctttga gcagatgcag
     5161 aactaatcta gataattcta ccgggtaggg gaggcgcttt tcccaaggca gtctggagca
     5221 tgcgctttag cagccccgct gggcacttgg cgctacacaa gtggcctctg gcctcgcaca
     5281 cattccacat ccaccggtag gcgccaaccg gctccgttct ttggtggccc cttcgcgcca
     5341 ccttctactc ctcccctagt caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg
     5401 acgtgacaaa tggaagtagc acgtctcact agtctcgtgc agatggacag caccgctgag
     5461 caatggaagc gggtaggcct ttggggcagc ggccaatagc agctttgctc cttcgctttc
     5521 tgggctcaga ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc
     5581 ggggcgggcg cccgaaggtc ctccggaggc ccggcattct gcacgcttca aaagcgcacg
     5641 tctgccgcgc tgttctcctc ttcctcatct ccgggccttt cgacctgcag cccaagctta
     5701 ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg
     5761 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg
     5821 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
     5881 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
     5941 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
     6001 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
     6061 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
     6121 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
     6181 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
     6241 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
     6301 gaccgcgtct ggaacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     6361 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     6421 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     6481 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     6541 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     6601 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     6661 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc
     6721 tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     6781 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     6841 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     6901 ccgcctggaa ttaattctgc agtcgagacc tagaaaaaca tggagcaatc acaagtagca
     6961 atacagcagc taccaatgct gattgtgcct ggctagaagc acaagaggag gaggaggtgg
     7021 gttttccagt cacacctcag gtacctttaa gaccaatgac ttacaaggca gctgtagatc
     7081 ttagccactt tttaaaagaa aagaggggac tggaagggct aattcactcc caacgaagac
     7141 aagatatcct tgatctgtgg atctaccaca cacaaggcta cttccctgat tagcagaact
     7201 acacaccagg gccaggggtc agatatccac tgacctttgg atggtgctac aagctagtac
     7261 cagttgagcc agataaggta gaagaggcca ataaaggaga gaacaccagc ttgttacacc
     7321 ctgtgagcct gcatgggatg gatgacccgg agagagaagt gttagagtgg aggtttgaca
     7381 gccgcctagc atttcatcac gtggcccgag agctgcatcc ggagtacttc aagaactgct
     7441 gatatcgagc ttgctacaag ggactttccg ctggggactt tccagggagg cgtggcctgg
     7501 gcgggactgg ggagtggcga gccctcagat cctgcatata agcagctgct ttttgcctgt
     7561 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     7621 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     7681 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     7741 agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata
     7801 tcagagagtg agaggccttg acattgctag cgttttaccg tcgacctcta gctagagctt
     7861 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     7921 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     7981 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     8041 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     8101 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     8161 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     8221 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     8281 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     8341 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     8401 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     8461 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     8521 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     8581 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     8641 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     8701 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     8761 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     8821 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     8881 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     8941 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     9001 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     9061 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     9121 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     9181 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     9241 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     9301 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     9361 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     9421 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     9481 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     9541 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     9601 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     9661 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     9721 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     9781 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     9841 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     9901 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     9961 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
    10021 acctgacgtc gacggatcgg gagatcaact tgtttattgc agcttataat ggttacaaat
    10081 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
    10141 gtttgtccaa actcatcaat gtatcttatc atgtctggat caactggata actcaagcta
    10201 accaaaatca tcccaaactt cccaccccat accctattac cactgccaat tacctgtggt
    10261 ttcatttact ctaaacctgt gattcctctg aattattttc attttaaaga aattgtattt
    10321 gttaaatatg tactacaaac ttagtagttt ttaaagaaat tgtatttgtt aaatatgtac
    10381 tacaaactta gtagt
//
