LOCUS       V032803                 8584 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032803
VERSION     V032803
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..3486
                     /label="DHODH(NM_001361)"
                     /note="DHODH(NM_001361)"
                     /gene="DHODH"
     CDS             3493..3510
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     promoter        3554..3765
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3778..4046
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4073..4672
                     /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    4679..5267
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5341..5574
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5646..5767
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5786..5921
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(5959..5975)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5983..5999
                     /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(6007..6037)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6052..6073
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6361..6949)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7120..7980)
                     /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(7981..8085)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8559..8575
                     /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 catggcgtgg agacacctga aaaagcgggc ccaggatgct
     2341 gtgatcatcc tggggggagg aggacttctc ttcgcctcct acctgatggc cacgggagat
     2401 gagcgtttct atgctgaaca cctgatgccg actctgcagg ggctgctgga cccggagtca
     2461 gcccacagac tggctgttcg cttcacctcc ctggggctcc ttccacgggc cagatttcaa
     2521 gactctgaca tgctggaagt gagagttctg ggccataaat tccgaaatcc agtaggaatt
     2581 gctgcaggat ttgacaagca tggggaagcc gtggacggac tttataagat gggctttggt
     2641 tttgttgaga taggaagtgt gactccaaaa cctcaggaag gaaaccctag acccagagtc
     2701 ttccgcctcc ctgaggacca agctgtcatt aacaggtatg gatttaacag tcacgggctt
     2761 tcagtggtgg aacacaggtt acgggccaga cagcagaagc aggccaagct cacagaagat
     2821 ggactgcctc tgggggtcaa cttggggaag aacaagacct cagtggacgc cgcggaggac
     2881 tacgcagaag gggtgcgcgt actgggcccc ctggccgact acctggtggt gaatgtgtcc
     2941 agccccaaca ctgccgggct gcggagcctt cagggaaagg ccgagctgcg ccgcctgctg
     3001 accaaggtgc tgcaggagag ggatggcttg cggagagtgc acaggccggc agtcctggtg
     3061 aagatcgctc ctgacctcac cagccaggat aaggaggaca ttgccagtgt ggtcaaagag
     3121 ttgggcatcg atgggctgat tgttacgaac accaccgtga gtcgccctgc gggcctccag
     3181 ggtgccctgc gctctgaaac aggagggctg agtgggaagc ccctccggga tttatcaact
     3241 caaaccattc gggagatgta tgcactcacc caaggccgag ttcccataat tggggttggt
     3301 ggtgtgagca gcgggcagga cgcgctggag aagatccggg caggggcctc cctggtgcag
     3361 ctgtacacgg ccctcacctt ctgggggcca cccgttgtgg gcaaagtcaa gcgggaactg
     3421 gaggcccttc tgaaagagca gggctttggc ggagtcacag atgccattgg agcagatcat
     3481 cggaggggat cccatcacca tcaccatcac tagcggccgc gaaggatctg cgatcgctcc
     3541 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
     3601 gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
     3661 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
     3721 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa gcttcgaggg
     3781 gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc acgccggttg
     3841 agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta
     3901 agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag cctacctaga
     3961 ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg
     4021 ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct agatgaccga
     4081 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg tacgcaccct
     4141 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg accgccacat
     4201 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     4261 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     4321 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     4381 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     4441 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     4501 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     4561 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     4621 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gagtcgacaa
     4681 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     4741 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     4801 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     4861 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     4921 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     4981 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     5041 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     5101 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     5161 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     5221 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa
     5281 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
     5341 tggaagggct aattcactcc caacgaaaat aagatctgct ttttgcttgt actgggtctc
     5401 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     5461 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     5521 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta
     5581 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg
     5641 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5701 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5761 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccagtt ccgcccattc
     5821 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     5881 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagactttt gcagagacgg
     5941 cccaaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     6001 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     6061 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     6121 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     6181 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     6241 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     6301 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     6361 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6421 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6481 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6541 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6601 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6661 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6721 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6781 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     6841 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     6901 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6961 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     7021 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     7081 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     7141 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     7201 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     7261 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     7321 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     7381 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     7441 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     7501 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     7561 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     7621 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     7681 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     7741 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     7801 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     7861 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     7921 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     7981 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     8041 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     8101 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     8161 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     8221 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     8281 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     8341 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     8401 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     8461 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     8521 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa
     8581 gctg
//
