LOCUS       V018731                 9146 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018731
VERSION     V018731
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             1..634
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     misc_feature    681..806
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1303..1536
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1721..1765
                     /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    2058..2175
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     enhancer        2227..2530
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2531..2734
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2853..4295
                     /label="OTUD1(NM_001145373)"
                     /note="OTUD1(NM_001145373)"
                     /gene="OTUD1"
     CDS             4314..4385
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags"
                     /transl_table=1
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
                     /label="3xFLAG"
     promoter        4403..4902
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4923..5522
                     /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    5536..6124
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6199..6432
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     promoter        6637..6741
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             6742..7602
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     rep_origin      7773..8361
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    8772..8793
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        8808..8838
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    8846..8862
                     /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"
     primer_bind     8870..8886
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtgga
     2221 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     2281 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     2341 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     2401 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     2461 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     2521 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     2581 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     2641 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     2701 gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaccgtca gatcgcctgg
     2761 agacgccatc cacgctgttt tgacctccat agaagacacc ggcggccacg cgtcgaagcc
     2821 cgggcggatc gggtttaaac ggatccgcca ccatgcagct ctacagcagc gtctgcaccc
     2881 actaccccgc cggggccccg ggtcccacgg ccgccgcccc cgcgccaccc gccgccgcta
     2941 cccccttcaa ggtctcgctg cagcccccgg gagccgccgg cgccgcgccc gagcccgaga
     3001 ccggtgagtg ccagcccgcc gcggccgccg agcaccggga agccgccgct gtccccgccg
     3061 ccaagatgcc cgccttctcc tcctgcttcg aggtggtgtc cggggccgcc gcgcccgcct
     3121 ccgccgccgc cggcccgccc ggcgcgtcct gcaagccgcc gctgccgccg cactacacgt
     3181 ccaccgcaca gatcaccgtg cgggccctgg gcgccgacag gctcctgctg cacgggcccg
     3241 atcccgttcc cggcgccgcg ggctccgccg ctgccccgcg cggccgctgc ctcctgctcg
     3301 ccccggcgcc cgcagccccg gtcccgccgc ggcggggctc ctcggcctgg ctcctggagg
     3361 agctgctgcg gcccgactgc cccgagcccg cgggcttgga cgcgacacgg gaggggcccg
     3421 atcggaactt ccgactgagc gagcaccgcc aggccctggc cgccgccaag caccgaggcc
     3481 ccgcggcgac cccggggagc cccgatcccg gccccggtcc gtggggcgaa gagcacttgg
     3541 cggagagggg ccccaggggc tgggagaggg gcggcgatcg ctgcgacgcg cccggtgggg
     3601 acgcggcgcg gaggcccgac ccagaggccg aggcaccccc cgccgggagc atcgaggccg
     3661 ccccgagtag tgcggcggag ccggtgatcg tctccaggtc ggatcccaga gacgagaagc
     3721 tggccctata cctggccgag gtggagaagc aggacaagta tctgcggcag aggaataagt
     3781 accgattcca catcattcca gacggcaact gcctctaccg agctgtcagc aagacggtgt
     3841 atggggacca gagcctgcac cgggagttga gggagcagac ggtgcactac atcgccgacc
     3901 atctcgacca cttcagcccc ctgattgagg gcgacgtggg ggagtttatc atcgctgctg
     3961 cccaagacgg ggcatgggcc gggtacccgg agttgctggc catggggcag atgctgaatg
     4021 tgaatatcca tttaactact ggagggaggc tggagagtcc cacggtgtct accatgattc
     4081 attatttggg cccagaggat tccctgaggc ctagtatttg gctcagttgg ctcagtaacg
     4141 gacactatga tgctgtattt gatcactcct atcctaaccc agagtacgac aactggtgca
     4201 aacaaactca agtgcaaagg aaacgcgacg aagaacttgc caaatctatg gccatatcct
     4261 tgtctaaaat gtatattgaa caaaatgcat gctctctcga gtctagaggg cccgattaca
     4321 aggatgacga cgataaggat tacaaggatg acgacgataa ggattacaag gatgacgacg
     4381 ataagtagtc gagaattcta ccgggtaggg gaggcgcttt tcccaaggca gtctggagca
     4441 tgcgctttag cagccccgct gggcacttgg cgctacacaa gtggcctctg gcctcgcaca
     4501 cattccacat ccaccggtag gcgccaaccg gctccgttct ttggtggccc cttcgcgcca
     4561 ccttctactc ctcccctagt caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg
     4621 acgtgacaaa tggaagtagc acgtctcact agtctcgtgc agatggacag caccgctgag
     4681 caatggaagc gggtaggcct ttggggcagc ggccaatagc agctttgctc cttcgctttc
     4741 tgggctcaga ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc
     4801 ggggcgggcg cccgaaggtc ctccggaggc ccggcattct gcacgcttca aaagcgcacg
     4861 tctgccgcgc tgttctcctc ttcctcatct ccgggccttt cgacctgcag cccaagctta
     4921 ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg
     4981 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg
     5041 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
     5101 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
     5161 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
     5221 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
     5281 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
     5341 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
     5401 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
     5461 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
     5521 gaatcgatag atcctaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     5581 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     5641 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     5701 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     5761 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     5821 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     5881 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     5941 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     6001 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     6061 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     6121 ccgcctgaga tcctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
     6181 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt
     6241 ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     6301 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     6361 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     6421 aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga
     6481 aatgaatatc agagagtgag aggcccgggt taattaagga aagggctaga tcattcttga
     6541 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     6601 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     6661 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     6721 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     6781 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     6841 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     6901 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     6961 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata
     7021 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     7081 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     7141 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     7201 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     7261 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     7321 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     7381 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     7441 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     7501 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     7561 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     7621 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     7681 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     7741 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     7801 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     7861 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     7921 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     7981 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     8041 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     8101 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     8161 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     8221 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     8281 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     8341 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     8401 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     8461 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     8521 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     8581 ccgattcatt aatgcagcaa gctcatggct gactaatttt ttttatttat gcagaggccg
     8641 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     8701 gcttttgcaa aaagctcccc gtggcacgac aggtttcccg actggaaagc gggcagtgag
     8761 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     8821 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     8881 tatgacatga ttacgaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     8941 ttgtccaaac tcatcaatgt atcttatcat gtctggatca actggataac tcaagctaac
     9001 caaaatcatc ccaaacttcc caccccatac cctattacca ctgccaatta cctgtggttt
     9061 catttactct aaacctgtga ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt
     9121 taaatatgta ctacaaactt agtagt
//
