LOCUS       V025666                 8777 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025666
VERSION     V025666
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             234..414
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    458..583
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1076..1309
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1493..1537
                     /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    1804..1920
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2010..2213
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2301..3626
                     /label="EDNRB(NM_001122659)"
                     /note="EDNRB(NM_001122659)"
                     /gene="EDNRB"
     CDS             3633..3698
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3742..3953
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3966..4234
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4266..4865
                     /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    4872..5460
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5534..5767
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5839..5960
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5979..6114
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6152..6168)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6176..6192
                     /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(6200..6230)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6245..6266
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6554..7142)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7313..8173)
                     /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(8174..8278)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8752..8768
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt
     1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag
     2221 tgaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagattcta
     2281 gagctagcga attcgccacc atgcagccgc ctccaagtct gtgcggacgc gccctggttg
     2341 cgctggttct tgcctgcggc ctgtcgcgga tctggggaga ggagagaggc ttcccgcctg
     2401 acagggccac tccgcttttg caaaccgcag agataatgac gccacccact aagaccttat
     2461 ggcccaaggg ttccaacgcc agtctggcgc ggtcgttggc acctgcggag gtgcctaaag
     2521 gagacaggac ggcaggatct ccgccacgca ccatctcccc tcccccgtgc caaggaccca
     2581 tcgagatcaa ggagactttc aaatacatca acacggttgt gtcctgcctt gtgttcgtgc
     2641 tggggatcat cgggaactcc acacttctga gaattatcta caagaacaag tgcatgcgaa
     2701 acggtcccaa tatcttgatc gccagcttgg ctctgggaga cctgctgcac atcgtcattg
     2761 acatccctat caatgtctac aagctgctgg cagaggactg gccatttgga gctgagatgt
     2821 gtaagctggt gcctttcata cagaaagcct ctgtgggaat cactgtgctg agtctatgtg
     2881 ctctgagtat tgacagatat cgagctgttg cttcttggag tagaattaaa ggaattgggg
     2941 ttccaaaatg gacagcagta gaaattgttt tgatttgggt ggtctctgtg gttctggctg
     3001 tccctgaagc cataggtttt gatataatta cgatggacta caaaggaagt tatctgcgaa
     3061 tctgcttgct tcatcccgtt cagaagacag ctttcatgca gttttacaag acagcaaaag
     3121 attggtggct attcagtttc tatttctgct tgccattggc catcactgca tttttttata
     3181 cactaatgac ctgtgaaatg ttgagaaaga aaagtggcat gcagattgct ttaaatgatc
     3241 acctaaagca gagacgggaa gtggccaaaa ccgtcttttg cctggtcctt gtctttgccc
     3301 tctgctggct tccccttcac ctcagcagga ttctgaagct cactctttat aatcagaatg
     3361 atcccaatag atgtgaactt ttgagctttc tgttggtatt ggactatatt ggtatcaaca
     3421 tggcttcact gaattcctgc attaacccaa ttgctctgta tttggtgagc aaaagattca
     3481 aaaactgctt taagtcatgc ttatgctgct ggtgccagtc atttgaagaa aaacagtcct
     3541 tggaggaaaa gcagtcgtgc ttaaagttca aagctaatga tcacggatat gacaacttcc
     3601 gttccagtaa taaatacagc tcatctggat ccgactacaa agaccatgac ggtgattata
     3661 aagatcatga catcgactac aaggatgacg atgacaagta gcggccgcga aggatctgcg
     3721 atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
     3781 ggggaggggt cggcaattga acgggtgcct agagaaggtg gcgcggggta aactgggaaa
     3841 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
     3901 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca cagctgaagc
     3961 ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc cgccatccac
     4021 gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg
     4081 tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc ccttggagcc
     4141 tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa ctctacgtct
     4201 ttgtttcgtt ttctgttctg cgccgttaca gatccaagct gtgaccggcg cctacgctag
     4261 ccaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg
     4321 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc
     4381 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
     4441 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
     4501 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
     4561 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
     4621 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
     4681 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
     4741 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
     4801 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg
     4861 cctgagtcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     4921 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     4981 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     5041 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     5101 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     5161 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     5221 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     5281 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     5341 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     5401 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     5461 ctggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     5521 gaaaaggggg gactggaagg gctaattcac tcccaacgaa aataagatct gctttttgct
     5581 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     5641 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     5701 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     5761 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga
     5821 atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     5881 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     5941 aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca
     6001 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     6061 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact
     6121 tttgcagaga cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     6181 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     6241 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6301 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6361 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6421 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6481 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6541 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6601 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6661 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6721 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     6781 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6841 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6901 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     6961 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     7021 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7081 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7141 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7201 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7261 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7321 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7381 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7441 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7501 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7561 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7621 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7681 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7741 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7801 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7861 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7921 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7981 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     8041 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     8101 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     8161 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8221 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8281 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     8341 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     8401 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     8461 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     8521 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     8581 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt
     8641 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     8701 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     8761 cggccagtgc caagctg
//
