LOCUS       V035665                 9137 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035665
VERSION     V035665
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..3318
                     /label="Ctsb(NM_007798)"
                     /note="Ctsb(NM_007798)"
                     /gene="Ctsb"
     CDS             3355..4065
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK*"
                     /label="mCherry"
     promoter        4107..4318
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4331..4599
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4626..5225
                     /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    5232..5820
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5894..6127
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6199..6320
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6339..6474
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6512..6528)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6536..6552
                     /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(6560..6590)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6605..6626
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6914..7502)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7673..8533)
                     /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(8534..8638)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9112..9128
                     /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 catgtggtgg tccttgatcc ttctttcttg cctgctggca
     2341 ctgaccagtg cccatgacaa gccttccttc cacccgctgt cggatgacct gattaactat
     2401 atcaacaaac agaatacaac atggcaggct ggacgcaact tctacaatgt tgacataagc
     2461 tatctgaaga agctgtgtgg cactgtcctg ggtggaccca aactgccagg aagggttgcg
     2521 ttcggtgagg acatagatct acctgaaacc tttgatgcac gggaacaatg gtccaactgc
     2581 ccgaccattg gacagattag agaccagggc tcctgcggct cttgttgggc atttggggca
     2641 gtggaagcca tttctgaccg aacctgcatt cacaccaatg gccgagtcaa cgtggaggtg
     2701 tctgctgaag acctgcttac ttgctgtggt atccagtgtg gggacggctg taatggtggc
     2761 tatccctctg gagcatggag cttctggaca aaaaaaggcc tggtttcagg tggagtctac
     2821 aattctcatg taggctgctt accatacacc atccctccct gcgagcacca tgtcaatggc
     2881 tcccgtcccc catgcactgg agaaggagat actcccaggt gcaacaagag ctgtgaagct
     2941 ggctactccc catcctacaa agaggataag cactttgggt acacttccta cagcgtgtct
     3001 aacagtgtga aggagatcat ggcagaaatc tacaaaaatg gcccagtgga gggtgccttc
     3061 actgtgtttt ctgacttctt gacttacaaa tcaggagtat acaagcatga agccggtgat
     3121 atgatgggtg gccacgccat ccgcatcctg ggctggggag tagagaatgg agttccctac
     3181 tggctggcag ccaactcttg gaaccttgac tggggtgata atggcttctt taaaatcctc
     3241 agaggagaga accactgtgg cattgaatca gaaattgtgg ctggaatccc acgcactgac
     3301 cagtactggg gaagattcgg atccggtgga ggcggatcag gaggcggtgg atctatggtg
     3361 agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac
     3421 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     3481 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc
     3541 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc
     3601 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
     3661 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
     3721 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
     3781 aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg
     3841 aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc
     3901 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc
     3961 aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc
     4021 gagggccgcc actccaccgg cggcatggac gagctgtaca agtaggcggc cgcgaaggat
     4081 ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa
     4141 gttgggggga ggggtcggca attgaacggg tgcctagaga aggtggcgcg gggtaaactg
     4201 ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat
     4261 aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagct
     4321 gaagcttcga ggggctcgca tctctccttc acgcgcccgc cgccctacct gaggccgcca
     4381 tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc
     4441 cgccgtctag gtaagtttaa agctcaggtc gagaccgggc ctttgtccgg cgctcccttg
     4501 gagcctacct agactcagcc ggctctccac gctttgcctg accctgcttg ctcaactcta
     4561 cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc aagctgtgac cggcgcctac
     4621 gctagatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg
     4681 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgacc
     4741 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
     4801 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
     4861 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
     4921 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
     4981 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
     5041 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
     5101 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
     5161 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg
     5221 cctgagtcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     5281 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     5341 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     5401 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     5461 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     5521 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     5581 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     5641 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     5701 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     5761 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     5821 ctggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     5881 gaaaaggggg gactggaagg gctaattcac tcccaacgaa aataagatct gctttttgct
     5941 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     6001 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     6061 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     6121 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga
     6181 atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     6241 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     6301 aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca
     6361 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     6421 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact
     6481 tttgcagaga cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     6541 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     6601 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6661 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6721 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6781 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6841 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6901 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6961 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     7021 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     7081 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     7141 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     7201 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     7261 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7321 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     7381 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7441 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7501 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7561 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7621 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7681 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7741 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7801 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7861 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7921 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7981 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     8041 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     8101 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     8161 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     8221 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     8281 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     8341 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     8401 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     8461 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     8521 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8581 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8641 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     8701 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     8761 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     8821 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     8881 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     8941 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt
     9001 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     9061 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     9121 cggccagtgc caagctg
//
