LOCUS       V018280                 9337 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018280
VERSION     V018280
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"
     regulatory      2809..2818
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             2818..2847
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             2854..4020
                     /label="Kctd9(NM_001108871)"
                     /note="Kctd9(NM_001108871)"
                     /gene="Kctd9"
     misc_feature    4040..4626
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES2"
     CDS             4646..5245
                     /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    5259..5847
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6054..6687
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6815..6831)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6839..6855
                     /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(6863..6893)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6908..6929
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7217..7805)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7976..8836)
                     /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(8837..8941)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    8989..9123
                     /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 tctatttccg gtgaattcgc caccatggaa
     2821 caaaaactca tctcagaaga ggatctgctc gagatgcggc gggtcacgct gttcttgaac
     2881 ggcagcccga ggaacggcaa ggtggttgct gtatacggaa ctttgtctga tttgctctca
     2941 gtagccagca gtaaactcgg cataaaagcc accagtgtat ataatgggaa aggcggactg
     3001 attgatgata ttgctttgat cagggatgat gatgtgttgt ttgtgtgtga aggagaacca
     3061 tttattgatc ctcagacaga tgccagaccg cctggaggat tgctaggatc tcacacagac
     3121 tggcttacat tgaatgtagg agggcgatac tttacaacaa caaggagcac tttagtgaat
     3181 aaagagcctg acagtatgct ggcgcacatg ttcaaggaca aaggtgtctg gggaaataag
     3241 caagatcata gaggagcttt cttaattgac cgaagtcctg agtactttga acctattttg
     3301 aactatttac gtcatggaca gctcattgta aatgatggca ttaacttatt gggtgttcta
     3361 gaagaagcca ggttttttgg tattgactca ctgattgaac acctagaagt ggccataaag
     3421 aattcccaac caccagagga tcattcacca atatcccgaa aggaatttgt tcgatttctg
     3481 ctggccactc caaccaagtc agagctgcgc tgccagggtt taaacttcag tggtgctgat
     3541 ctttcccgtt tggaccttcg atacattaac ttcaaaatgg ccaatttaag ccgctgcaac
     3601 ctcgcacatg caaatctctg ctgtgctaat cttgaacgag ctgatctttc tggatcagtg
     3661 ctcgactgtg caaatctcca gggagtcaag atgctctgct caaacgcaga aggtgcatcc
     3721 ctgaggctct gtaattttga ggatccttca ggtcttaaag ccaacttaga gggtgctaat
     3781 ctgaaaggtg tggacatgga aggaagccag atgacaggga ttaacctcag agttgctacc
     3841 ttaaaaaatg cgaaactgaa gaactgtaat ctcagaggag caactctggc aggaacagat
     3901 ttagaaaact gtgatttgtc ggggtgtgac ctccaggaag ccaacttgag aggctccaac
     3961 gtgaagggtg ccatatttga agagatgctg acaccattac atatgtctca gagtgtcaga
     4021 tgagcggccg cggatcccgc ccctctccct cccccccccc taacgttact ggccgaagcc
     4081 gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt
     4141 ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc
     4201 tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc
     4261 tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc
     4321 cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg
     4381 cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct
     4441 cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat
     4501 ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc
     4561 taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga taagcttgcc
     4621 acaacccaca aggagacgac cttccatgac cgagtacaag cccacggtgc gcctcgccac
     4681 ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     4741 cacgcgccac accgtcgacc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     4801 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     4861 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     4921 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     4981 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     5041 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     5101 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     5161 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     5221 catgacccgc aagcccggtg cctagacgcg tctggaacaa tcaacctctg gattacaaaa
     5281 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     5341 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     5401 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     5461 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     5521 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     5581 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     5641 tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc
     5701 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     5761 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     5821 ggatctccct ttgggccgcc tccccgcctg gaattaattc tgcagtcgag acctagaaaa
     5881 acatggagca atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga
     5941 agcacaagag gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat
     6001 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaagaggg gactggaagg
     6061 gctaattcac tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg
     6121 ctacttccct gattagcaga actacacacc agggccaggg gtcagatatc cactgacctt
     6181 tggatggtgc tacaagctag taccagttga gccagataag gtagaagagg ccaataaagg
     6241 agagaacacc agcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga
     6301 agtgttagag tggaggtttg acagccgcct agcatttcat cacgtggccc gagagctgca
     6361 tccggagtac ttcaagaact gctgatatcg agcttgctac aagggacttt ccgctgggga
     6421 ctttccaggg aggcgtggcc tgggcgggac tggggagtgg cgagccctca gatcctgcat
     6481 ataagcagct gctttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg
     6541 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     6601 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     6661 tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt
     6721 tataacttgc aaagaaatga atatcagaga gtgagaggcc ttgacattgc tagcgtttac
     6781 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     6841 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     6901 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     6961 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     7021 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     7081 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     7141 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     7201 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     7261 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     7321 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     7381 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     7441 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     7501 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     7561 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     7621 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     7681 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     7741 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     7801 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     7861 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     7921 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     7981 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     8041 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     8101 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     8161 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     8221 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     8281 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     8341 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     8401 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     8461 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     8521 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     8581 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     8641 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     8701 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     8761 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     8821 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     8881 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     8941 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatcaa cttgtttatt
     9001 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     9061 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     9121 atcaactgga taactcaagc taaccaaaat catcccaaac ttcccacccc ataccctatt
     9181 accactgcca attacctgtg gtttcattta ctctaaacct gtgattcctc tgaattattt
     9241 tcattttaaa gaaattgtat ttgttaaata tgtactacaa acttagtagt ttttaaagaa
     9301 attgtatttg ttaaatatgt actacaaact tagtagt
//
