LOCUS       V020035                 8867 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020035
VERSION     V020035
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    1798..1915
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2005..2208
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2297..3787
                     /label="MDM2(NM_002392)"
                     /note="MDM2(NM_002392)"
                     /gene="MDM2"
     promoter        3837..4048
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4061..4329
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4356..4955
                     /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    4962..5550
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5624..5857
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5929..6050
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6069..6204
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6242..6258)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6266..6282
                     /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(6290..6320)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6335..6356
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6644..7232)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7403..8263)
                     /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(8264..8368)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8842..8858
                     /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 ggttaacttt
     1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga
     1921 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
     2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc
     2281 tagcgaattc gccaccatgg tgaggagcag gcaaatgtgc aataccaaca tgtctgtacc
     2341 tactgatggt gctgtaacca cctcacagat tccagcttcg gaacaagaga ccctggttag
     2401 accaaagcca ttgcttttga agttattaaa gtctgttggt gcacaaaaag acacttatac
     2461 tatgaaagag gttctttttt atcttggcca gtatattatg actaaacgat tatatgatga
     2521 gaagcaacaa catattgtat attgttcaaa tgatcttcta ggagatttgt ttggcgtgcc
     2581 aagcttctct gtgaaagagc acaggaaaat atataccatg atctacagga acttggtagt
     2641 agtcaatcag caggaatcat cggactcagg tacatctgtg agtgagaaca ggtgtcacct
     2701 tgaaggtggg agtgatcaaa aggaccttgt acaagagctt caggaagaga aaccttcatc
     2761 ttcacatttg gtttctagac catctacctc atctagaagg agagcaatta gtgagacaga
     2821 agaaaattca gatgaattat ctggtgaacg acaaagaaaa cgccacaaat ctgatagtat
     2881 ttccctttcc tttgatgaaa gcctggctct gtgtgtaata agggagatat gttgtgaaag
     2941 aagcagtagc agtgaatcta cagggacgcc atcgaatccg gatcttgatg ctggtgtaag
     3001 tgaacattca ggtgattggt tggatcagga ttcagtttca gatcagttta gtgtagaatt
     3061 tgaagttgaa tctctcgact cagaagatta tagccttagt gaagaaggac aagaactctc
     3121 agatgaagat gatgaggtat atcaagttac tgtgtatcag gcaggggaga gtgatacaga
     3181 ttcatttgaa gaagatcctg aaatttcctt agctgactat tggaaatgca cttcatgcaa
     3241 tgaaatgaat cccccccttc catcacattg caacagatgt tgggcccttc gtgagaattg
     3301 gcttcctgaa gataaaggga aagataaagg ggaaatctct gagaaagcca aactggaaaa
     3361 ctcaacacaa gctgaagagg gctttgatgt tcctgattgt aaaaaaacta tagtgaatga
     3421 ttccagagag tcatgtgttg aggaaaatga tgataaaatt acacaagctt cacaatcaca
     3481 agaaagtgaa gactattctc agccatcaac ttctagtagc attatttata gcagccaaga
     3541 agatgtgaaa gagtttgaaa gggaagaaac ccaagacaaa gaagagagtg tggaatctag
     3601 tttgcccctt aatgccattg aaccttgtgt gatttgtcaa ggtcgaccta aaaatggttg
     3661 cattgtccat ggcaaaacag gacatcttat ggcctgcttt acatgtgcaa agaagctaaa
     3721 gaaaaggaat aagccctgcc cagtatgtag acaaccaatt caaatgattg tgctaactta
     3781 tttcccctag gatccgcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc
     3841 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg
     3901 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
     3961 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
     4021 tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc
     4081 acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc
     4141 tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc
     4201 gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac
     4261 gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg
     4321 ttacagatcc aagctgtgac cggcgcctac gctagatgac cgagtacaag cccacggtgc
     4381 gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc gcgttcgccg
     4441 actaccccgc cacgcgccac accgtcgacc cggaccgcca catcgagcgg gtcaccgagc
     4501 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
     4561 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
     4621 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
     4681 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     4741 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     4801 aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
     4861 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     4921 gcacctggtg catgacccgc aagcccggtg cctgagtcga caatcaacct ctggattaca
     4981 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     5041 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     5101 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     5161 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     5221 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     5281 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     5341 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga
     5401 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     5461 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     5521 gtcggatctc cctttgggcc gcctccccgc ctggtacctt taagaccaat gacttacaag
     5581 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac
     5641 tcccaacgaa aataagatct gctttttgct tgtactgggt ctctctggtt agaccagatc
     5701 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     5761 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     5821 ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt
     5881 attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt
     5941 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     6001 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     6061 ctctagctat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     6121 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     6181 agtgaggagg cttttttgga ggcctagact tttgcagaga cggcccaaat tcgtaatcat
     6241 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     6301 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     6361 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     6421 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     6481 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6541 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6601 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6661 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6721 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6781 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6841 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6901 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6961 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     7021 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     7081 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     7141 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7201 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7261 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7321 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7381 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7441 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     7501 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     7561 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7621 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7681 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     7741 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7801 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7861 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7921 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7981 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     8041 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     8101 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     8161 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     8221 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8281 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8341 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     8401 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     8461 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     8521 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     8581 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     8641 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     8701 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     8761 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     8821 ggttttccca gtcacgacgt tgtaaaacga cggccagtgc caagctg
//
