LOCUS       V023974                 9198 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023974
VERSION     V023974
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    2028..2143
                     /note="central polypurine tract and central termination
                     sequence of HIV-1 (lacking the first T)"
                     /label="cPPT/CTS"
     enhancer        2201..2504
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2505..2708
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2826..3833
                     /label="RNF2(NM_007212)"
                     /note="RNF2(NM_007212)"
                     /gene="RNF2"
     CDS             3840..3929
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     promoter        3986..4485
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4506..5105
                     /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    5119..5707
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5914..6547
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6676..6692)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6700..6716
                     /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(6724..6754)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6769..6790
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7078..7666)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7837..8697)
                     /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(8698..8802)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    8850..8984
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
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 tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa
     2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     2761 tccatagaag acaccgactc tactagagga tcgctagcgc taccggactc agatctcgag
     2821 ccaccatgtc tcaggctgtg cagacaaacg gaactcaacc attaagcaaa acatgggaac
     2881 tcagtttata tgagttacaa cgaacacctc aggaggcaat aacagatggc ttagaaattg
     2941 tggtttcacc tcgaagtcta cacagtgaat taatgtgccc aatttgtttg gatatgttga
     3001 agaacaccat gactacaaag gagtgtttac atcgtttttg tgcagactgc atcatcacag
     3061 cccttagaag tggcaacaaa gaatgtccta cctgtcggaa aaaactagtt tccaaaagat
     3121 cactaaggcc agacccaaac tttgatgcac tcatcagcaa aatttatcca agtcgtgatg
     3181 agtatgaagc tcatcaagag agagtattag ccaggatcaa caagcacaat aatcagcaag
     3241 cactcagtca cagcattgag gaaggactga agatacaggc catgaacaga ctgcagcgag
     3301 gcaagaaaca acagattgaa aatggtagtg gagcagaaga taatggtgac agttcacact
     3361 gcagtaatgc atccacacat agcaatcagg aagcaggccc tagtaacaaa cggaccaaaa
     3421 catctgatga ttctgggcta gagcttgata ataacaatgc agcaatggca attgatccag
     3481 taatggatgg tgctagtgaa attgaattag tattcaggcc tcatcccaca cttatggaaa
     3541 aagatgacag tgcacagacg agatacataa agacttctgg taacgccact gttgatcact
     3601 tatccaagta tctggctgtg aggttagctt tagaagaact tcgaagcaaa ggtgaatcaa
     3661 accagatgaa ccttgataca gccagtgaga agcagtatac catttatata gcaacagcca
     3721 gtggccagtt cactgtatta aatggctctt tttctttgga attggtcagt gagaaatact
     3781 ggaaagtgaa caaacccatg gaactttatt acgcacctac aaaggagcac aaagaattcg
     3841 aacaaaaact aatatcagag gaagatttgg aacagaaact aatcagtgaa gaagatttag
     3901 agcaaaagtt aatttccgag gaggacttat agtgcagtcg acggtaccgc gggcccggga
     3961 tcccgcgact ctagataatt ctaccgggta ggggaggcgc ttttcccaag gcagtctgga
     4021 gcatgcgctt tagcagcccc gctgggcact tggcgctaca caagtggcct ctggcctcgc
     4081 acacattcca catccaccgg taggcgccaa ccggctccgt tctttggtgg ccccttcgcg
     4141 ccaccttcta ctcctcccct agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc
     4201 aggacgtgac aaatggaagt agcacgtctc actagtctcg tgcagatgga cagcaccgct
     4261 gagcaatgga agcgggtagg cctttggggc agcggccaat agcagctttg ctccttcgct
     4321 ttctgggctc agaggctggg aaggggtggg tccgggggcg ggctcagggg cgggctcagg
     4381 ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat tctgcacgct tcaaaagcgc
     4441 acgtctgccg cgctgttctc ctcttcctca tctccgggcc tttcgacctg cagcccaagc
     4501 ttaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg
     4561 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc
     4621 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
     4681 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
     4741 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
     4801 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
     4861 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
     4921 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
     4981 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
     5041 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg
     5101 cctgaccgcg tctggaacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     5161 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     5221 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     5281 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     5341 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     5401 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     5461 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg
     5521 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     5581 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     5641 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     5701 tccccgcctg gaattaattc tgcagtcgag acctagaaaa acatggagca atcacaagta
     5761 gcaatacagc agctaccaat gctgattgtg cctggctaga agcacaagag gaggaggagg
     5821 tgggttttcc agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag
     5881 atcttagcca ctttttaaaa gaaaagaggg gactggaagg gctaattcac tcccaacgaa
     5941 gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct gattagcaga
     6001 actacacacc agggccaggg gtcagatatc cactgacctt tggatggtgc tacaagctag
     6061 taccagttga gccagataag gtagaagagg ccaataaagg agagaacacc agcttgttac
     6121 accctgtgag cctgcatggg atggatgacc cggagagaga agtgttagag tggaggtttg
     6181 acagccgcct agcatttcat cacgtggccc gagagctgca tccggagtac ttcaagaact
     6241 gctgatatcg agcttgctac aagggacttt ccgctgggga ctttccaggg aggcgtggcc
     6301 tgggcgggac tggggagtgg cgagccctca gatcctgcat ataagcagct gctttttgcc
     6361 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     6421 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     6481 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     6541 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga
     6601 atatcagaga gtgagaggcc ttgacattgc tagcgtttta ccgtcgacct ctagctagag
     6661 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     6721 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     6781 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     6841 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     6901 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     6961 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     7021 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     7081 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     7141 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     7201 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     7261 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     7321 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     7381 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     7441 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     7501 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     7561 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     7621 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     7681 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     7741 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     7801 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     7861 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     7921 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     7981 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     8041 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     8101 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     8161 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     8221 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     8281 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     8341 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     8401 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     8461 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     8521 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     8581 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     8641 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     8701 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     8761 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     8821 gccacctgac gtcgacggat cgggagatca acttgtttat tgcagcttat aatggttaca
     8881 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     8941 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcaactgg ataactcaag
     9001 ctaaccaaaa tcatcccaaa cttcccaccc cataccctat taccactgcc aattacctgt
     9061 ggtttcattt actctaaacc tgtgattcct ctgaattatt ttcattttaa agaaattgta
     9121 tttgttaaat atgtactaca aacttagtag tttttaaaga aattgtattt gttaaatatg
     9181 tactacaaac ttagtagt
//
