LOCUS       V035971                 8507 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035971
VERSION     V035971
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             2302..2679
                     /label="H2BC21(NM_003528)"
                     /note="H2BC21(NM_003528)"
                     /gene="H2BC21"
     CDS             2716..3426
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="enhanced monomeric blue fluorescent protein
                     (Subach et al., 2011)"
                     /transl_table=1
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
                     /label="mTagBFP2"
     promoter        3477..3688
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3701..3969
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             3996..4595
                     /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    4602..5190
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5264..5497
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5569..5690
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5709..5844
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(5882..5898)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5906..5922
                     /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(5930..5960)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5975..5996
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6284..6872)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7043..7903)
                     /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(7904..8008)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8482..8498
                     /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 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct
     2281 agagctagcg aattcgccac catgcctgaa ccggcaaaat ccgctccggc ccctaaaaag
     2341 ggctccaaga aagccgtcac caaagcccag aagaaagacg gcaagaagcg caagcgcagc
     2401 cgcaaagaga gctactccat ctacgtgtac aaggtgctga agcaggtcca ccccgacacc
     2461 ggcatctcgt ccaaggccat gggcatcatg aactccttcg tcaacgacat cttcgagcgc
     2521 atcgcgggag aggcttcccg cctggcgcac tacaacaagc gctccaccat cacatcccgc
     2581 gagatccaga cggccgtgcg cctgctgctg cccggcgagc tggccaagca cgccgtgtcc
     2641 gagggcacca aggcggtcac caagtacacc agctccaagc tcgagggtgg aggcggatca
     2701 ggaggcggtg gatctatggt gtctaagggc gaagagctga ttaaggagaa catgcacatg
     2761 aagctgtaca tggagggcac cgtggacaac catcacttca agtgcacatc cgagggcgaa
     2821 ggcaagccct acgagggcac ccagaccatg agaatcaagg tggtcgaggg cggccctctc
     2881 cccttcgcct tcgacatcct ggctactagc ttcctctacg gcagcaagac cttcatcaac
     2941 cacacccagg gcatccccga cttcttcaag cagtccttcc ctgagggctt cacatgggag
     3001 agagtcacca catacgaaga cgggggcgtg ctgaccgcta cccaggacac cagcctccag
     3061 gacggctgcc tcatctacaa cgtcaagatc agaggggtga acttcacatc caacggccct
     3121 gtgatgcaga agaaaacact cggctgggag gccttcaccg agacgctgta ccccgctgac
     3181 ggcggcctgg aaggcagaaa cgacatggcc ctgaagctcg tgggcgggag ccatctgatc
     3241 gcaaacgcca agaccacata tagatccaag aaacccgcta agaacctcaa gatgcctggc
     3301 gtctactatg tggactacag actggaaaga atcaaggagg ccaacaacga gacctacgtc
     3361 gagcagcacg aggtggcagt ggccagatac tgcgacctcc ctagcaaact ggggcacaag
     3421 cttaattaag gatccgcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc
     3481 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg
     3541 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
     3601 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
     3661 tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc
     3721 acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc
     3781 tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc
     3841 gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac
     3901 gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg
     3961 ttacagatcc aagctgtgac cggcgcctac gctagatgac cgagtacaag cccacggtgc
     4021 gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc gcgttcgccg
     4081 actaccccgc cacgcgccac accgtcgacc cggaccgcca catcgagcgg gtcaccgagc
     4141 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
     4201 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
     4261 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
     4321 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     4381 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     4441 aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
     4501 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     4561 gcacctggtg catgacccgc aagcccggtg cctgagtcga caatcaacct ctggattaca
     4621 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     4681 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     4741 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     4801 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     4861 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     4921 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     4981 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga
     5041 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     5101 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     5161 gtcggatctc cctttgggcc gcctccccgc ctggtacctt taagaccaat gacttacaag
     5221 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac
     5281 tcccaacgaa aataagatct gctttttgct tgtactgggt ctctctggtt agaccagatc
     5341 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     5401 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     5461 ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt
     5521 attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt
     5581 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     5641 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     5701 ctctagctat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     5761 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     5821 agtgaggagg cttttttgga ggcctagact tttgcagaga cggcccaaat tcgtaatcat
     5881 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     5941 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     6001 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     6061 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     6121 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6181 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6241 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6301 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6361 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6421 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6481 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6541 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6601 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6661 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6721 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6781 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     6841 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6901 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6961 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7021 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7081 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     7141 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     7201 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7261 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7321 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     7381 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7441 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7501 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7561 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7621 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     7681 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7741 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7801 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     7861 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     7921 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     7981 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     8041 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     8101 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     8161 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     8221 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     8281 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     8341 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     8401 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     8461 ggttttccca gtcacgacgt tgtaaaacga cggccagtgc caagctg
//
