LOCUS       V022710                 9522 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022710
VERSION     V022710
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..4176
                     /label="MYC(NM_002467)"
                     /note="MYC(NM_002467)"
                     /gene="MYC"
     CDS             4177..4194
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     misc_feature    4225..4811
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES2"
     CDS             4831..5430
                     /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    5444..6032
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6239..6872
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7000..7016)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7024..7040
                     /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(7048..7078)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7093..7114
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7402..7990)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8161..9021)
                     /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(9022..9126)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9174..9308
                     /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 caccatggat
     2821 ttttttcggg tagtggaaaa ccagcagcct cccgcgacga tgcccctcaa cgttagcttc
     2881 accaacagga actatgacct cgactacgac tcggtgcagc cgtatttcta ctgcgacgag
     2941 gaggagaact tctaccagca gcagcagcag agcgagctgc agcccccggc gcccagcgag
     3001 gatatctgga agaaattcga gctgctgccc accccgcccc tgtcccctag ccgccgctcc
     3061 gggctctgct cgccctccta cgttgcggtc acacccttct cccttcgggg agacaacgac
     3121 ggcggtggcg ggagcttctc cacggccgac cagctggaga tggtgaccga gctgctggga
     3181 ggagacatgg tgaaccagag tttcatctgc gacccggacg acgagacctt catcaaaaac
     3241 atcatcatcc aggactgtat gtggagcggc ttctcggccg ccgccaagct cgtctcagag
     3301 aagctggcct cctaccaggc tgcgcgcaaa gacagcggca gcccgaaccc cgcccgcggc
     3361 cacagcgtct gctccacctc cagcttgtac ctgcaggatc tgagcgccgc cgcctcagag
     3421 tgcatcgacc cctcggtggt cttcccctac cctctcaacg acagcagctc gcccaagtcc
     3481 tgcgcctcgc aagactccag cgccttctct ccgtcctcgg attctctgct ctcctcgacg
     3541 gagtcctccc cgcagggcag ccccgagccc ctggtgctcc atgaggagac accgcccacc
     3601 accagcagcg actctgagga ggaacaagaa gatgaggaag aaatcgatgt tgtttctgtg
     3661 gaaaagaggc aggctcctgg caaaaggtca gagtctggat caccttctgc tggaggccac
     3721 agcaaacctc ctcacagccc actggtcctc aagaggtgcc acgtctccac acatcagcac
     3781 aactacgcag cgcctccctc cactcggaag gactatcctg ctgccaagag ggtcaagttg
     3841 gacagtgtca gagtcctgag acagatcagc aacaaccgaa aatgcaccag ccccaggtcc
     3901 tcggacaccg aggagaatgt caagaggcga acacacaacg tcttggagcg ccagaggagg
     3961 aacgagctaa aacggagctt ttttgccctg cgtgaccaga tcccggagtt ggaaaacaat
     4021 gaaaaggccc ccaaggtagt tatccttaaa aaagccacag catacatcct gtccgtccaa
     4081 gcagaggagc aaaagctcat ttctgaagag gacttgttgc ggaaacgacg agaacagttg
     4141 aaacacaaac ttgaacagct acggaactct tgtgcgcatc atcaccatca ccattaacta
     4201 gttctagagc ggccgcggat cccgcccctc tccctccccc ccccctaacg ttactggccg
     4261 aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc
     4321 gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag
     4381 gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt
     4441 tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa
     4501 ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc
     4561 aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg
     4621 gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat
     4681 gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa
     4741 acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataagc
     4801 ttgccacaac ccacaaggag acgaccttcc atgaccgagt acaagcccac ggtgcgcctc
     4861 gccacccgcg acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt cgccgactac
     4921 cccgccacgc gccacaccgt cgacccggac cgccacatcg agcgggtcac cgagctgcaa
     4981 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     5041 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     5101 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa
     5161 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc
     5221 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg
     5281 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
     5341 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     5401 tggtgcatga cccgcaagcc cggtgcctag acgcgtctgg aacaatcaac ctctggatta
     5461 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     5521 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     5581 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     5641 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     5701 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     5761 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     5821 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     5881 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     5941 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     6001 gagtcggatc tccctttggg ccgcctcccc gcctggaatt aattctgcag tcgagaccta
     6061 gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga ttgtgcctgg
     6121 ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt acctttaaga
     6181 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa gaggggactg
     6241 gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat ctaccacaca
     6301 caaggctact tccctgatta gcagaactac acaccagggc caggggtcag atatccactg
     6361 acctttggat ggtgctacaa gctagtacca gttgagccag ataaggtaga agaggccaat
     6421 aaaggagaga acaccagctt gttacaccct gtgagcctgc atgggatgga tgacccggag
     6481 agagaagtgt tagagtggag gtttgacagc cgcctagcat ttcatcacgt ggcccgagag
     6541 ctgcatccgg agtacttcaa gaactgctga tatcgagctt gctacaaggg actttccgct
     6601 ggggactttc cagggaggcg tggcctgggc gggactgggg agtggcgagc cctcagatcc
     6661 tgcatataag cagctgcttt ttgcctgtac tgggtctctc tggttagacc agatctgagc
     6721 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     6781 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     6841 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca
     6901 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggccttgac attgctagcg
     6961 tttaccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     7021 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     7081 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     7141 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     7201 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     7261 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     7321 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     7381 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     7441 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     7501 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     7561 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     7621 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     7681 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     7741 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     7801 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     7861 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     7921 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     7981 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     8041 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     8101 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     8161 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     8221 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     8281 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     8341 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     8401 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     8461 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     8521 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     8581 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     8641 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     8701 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     8761 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     8821 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     8881 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     8941 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     9001 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     9061 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     9121 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atcaacttgt
     9181 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     9241 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     9301 tctggatcaa ctggataact caagctaacc aaaatcatcc caaacttccc accccatacc
     9361 ctattaccac tgccaattac ctgtggtttc atttactcta aacctgtgat tcctctgaat
     9421 tattttcatt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagttttta
     9481 aagaaattgt atttgttaaa tatgtactac aaacttagta gt
//
