LOCUS       V021147                10360 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021147
VERSION     V021147
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             2815..5004
                     /label="DTL(NM_016448)"
                     /note="DTL(NM_016448)"
                     /gene="DTL"
     CDS             5005..5031
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    5063..5649
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES2"
     CDS             5669..6268
                     /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    6282..6870
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             7077..7710
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7838..7854)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7862..7878
                     /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(7886..7916)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7931..7952
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8240..8828)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8999..9859)
                     /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(9860..9964)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    10012..10146
                     /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 caccatgctc
     2821 ttcaattcgg tgctccgcca gccccagctt ggcgtcctga gaaatggatg gtcttcacaa
     2881 taccctcttc aatcccttct gactggttat cagtgcagtg gtaatgatga acacacttct
     2941 tatggagaaa caggagtccc agttcctcct tttggatgta ccttctcttc tgctcccaat
     3001 atggaacatg tactagcagt tgccaatgaa gaaggctttg ttcgattgta taacacagaa
     3061 tcacaaagtt tcagaaagaa gtgcttcaaa gaatggatgg ctcactggaa tgccgtcttt
     3121 gacctggcct gggttcctgg tgaacttaaa cttgttacag cagcaggtga tcaaacagcc
     3181 aaattttggg acgtaaaagc tggtgagctg attggaacat gcaaaggtca tcaatgcagc
     3241 ctcaagtcag ttgccttttc taagtttgag aaagctgtat tctgtacggg tggaagagat
     3301 ggcaacatta tggtctggga taccaggtgc aacaaaaaag atgggtttta taggcaagtg
     3361 aatcaaatca gtggagctca caatacctca gacaagcaaa ccccttcaaa acccaagaag
     3421 aaacagaatt caaaaggact tgctccttct gtggatttcc agcaaagtgt tactgtggtc
     3481 ctctttcaag acgagaatac cttagtctca gcaggagctg tggatgggat aatcaaagta
     3541 tgggatttac gtaagaatta tactgcttat cgacaagaac ccatagcatc caagtctttc
     3601 ctgtacccag gtagcagcac tcgaaaactt ggatattcaa gtctgatttt ggattccact
     3661 ggctctactt tatttgctaa ttgcacagac gataacatct acatgtttaa tatgactggg
     3721 ttgaagactt ctccagtggc tattttcaat ggacaccaga actctacctt ttatgtaaaa
     3781 tccagcctta gtccagatga ccagttttta gtcagtggct caagtgatga agctgcctac
     3841 atatggaagg tctccacacc ctggcaacct cctactgtgc tcctgggtca ttctcaagag
     3901 gtcacgtctg tgtgctggtg tccatctgac ttcacaaaga ttgctacctg ttctgatgac
     3961 aatacactaa aaatctggcg cttgaataga ggcttagagg agaaaccagg aggtgataaa
     4021 ctttccacgg tgggttgggc ctctcagaag aaaaaagagt caagacctgg cctagtaaca
     4081 gtaacgagta gccagagtac tcctgccaaa gcccccaggg caaagtgcaa tccatccaat
     4141 tcttccccgt catccgcagc ttgtgcccca agctgtgctg gagacctccc tcttccttca
     4201 aatactccta cgttctctat taaaacctct cctgccaagg cccggtctcc catcaacaga
     4261 agaggctctg tctcctccgt ctctcccaag ccaccttcat ctttcaagat gtcgattaga
     4321 aactgggtga cccgaacacc ttcctcatca ccacccatca ctccacctgc ttcggagacc
     4381 aagatcatgt ctccgagaaa agcccttatt cctgtgagcc agaagtcatc ccaagcagag
     4441 gcttgctctg agtctagaaa tagagtaaag aggaggctag actcaagctg tctggagagt
     4501 gtgaaacaaa agtgtgtgaa gagttgtaac tgtgtgactg agcttgatgg ccaagttgaa
     4561 aatcttcatt tggatctgtg ctgccttgct ggtaaccagg aagaccttag taaggactct
     4621 ctaggtccta ccaaatcaag caaaattgaa ggagctggta ccagtatctc agagcctccg
     4681 tctcctatca gtccgtatgc ttcagaaagc tgtggaacgc tacctcttcc tttgagacct
     4741 tgtggagaag ggtctgaaat ggtaggcaaa gagaatagtt ccccagagaa taaaaactgg
     4801 ttgttggcca tggcagccaa acggaaggct gagaatccat ctccacgaag tccgtcatcc
     4861 cagacaccca attccaggag acagagcgga aagaaattgc caagcccggt caccatcacg
     4921 cccagctcca tgaggaaaat ctgcacatac ttccatagaa agtcccagga ggacttctgt
     4981 ggtcctgaac actcaacaga attataccca tacgacgtac cagattacgc ttagactagt
     5041 tctagagcgg ccgcggatcc cgcccctctc cctccccccc ccctaacgtt actggccgaa
     5101 gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt
     5161 cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg
     5221 gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc
     5281 ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc
     5341 ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa
     5401 aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc
     5461 tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg
     5521 gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac
     5581 gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataagctt
     5641 gccacaaccc acaaggagac gaccttccat gaccgagtac aagcccacgg tgcgcctcgc
     5701 cacccgcgac gacgtccccc gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     5761 cgccacgcgc cacaccgtcg acccggaccg ccacatcgag cgggtcaccg agctgcaaga
     5821 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     5881 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     5941 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     6001 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     6061 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     6121 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     6181 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     6241 gtgcatgacc cgcaagcccg gtgcctagac gcgtctggaa caatcaacct ctggattaca
     6301 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     6361 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     6421 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     6481 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     6541 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     6601 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     6661 tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga
     6721 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     6781 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     6841 gtcggatctc cctttgggcc gcctccccgc ctggaattaa ttctgcagtc gagacctaga
     6901 aaaacatgga gcaatcacaa gtagcaatac agcagctacc aatgctgatt gtgcctggct
     6961 agaagcacaa gaggaggagg aggtgggttt tccagtcaca cctcaggtac ctttaagacc
     7021 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaaga ggggactgga
     7081 agggctaatt cactcccaac gaagacaaga tatccttgat ctgtggatct accacacaca
     7141 aggctacttc cctgattagc agaactacac accagggcca ggggtcagat atccactgac
     7201 ctttggatgg tgctacaagc tagtaccagt tgagccagat aaggtagaag aggccaataa
     7261 aggagagaac accagcttgt tacaccctgt gagcctgcat gggatggatg acccggagag
     7321 agaagtgtta gagtggaggt ttgacagccg cctagcattt catcacgtgg cccgagagct
     7381 gcatccggag tacttcaaga actgctgata tcgagcttgc tacaagggac tttccgctgg
     7441 ggactttcca gggaggcgtg gcctgggcgg gactggggag tggcgagccc tcagatcctg
     7501 catataagca gctgcttttt gcctgtactg ggtctctctg gttagaccag atctgagcct
     7561 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     7621 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     7681 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt
     7741 atttataact tgcaaagaaa tgaatatcag agagtgagag gccttgacat tgctagcgtt
     7801 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     7861 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     7921 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     7981 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     8041 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     8101 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     8161 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     8221 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     8281 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     8341 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     8401 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     8461 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     8521 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     8581 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     8641 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     8701 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     8761 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     8821 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     8881 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     8941 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     9001 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     9061 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     9121 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     9181 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     9241 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     9301 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     9361 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     9421 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     9481 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     9541 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     9601 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     9661 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     9721 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     9781 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     9841 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     9901 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     9961 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat caacttgttt
    10021 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
    10081 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
    10141 tggatcaact ggataactca agctaaccaa aatcatccca aacttcccac cccataccct
    10201 attaccactg ccaattacct gtggtttcat ttactctaaa cctgtgattc ctctgaatta
    10261 ttttcatttt aaagaaattg tatttgttaa atatgtacta caaacttagt agtttttaaa
    10321 gaaattgtat ttgttaaata tgtactacaa acttagtagt
//
