LOCUS       V046916                 9715 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046916
VERSION     V046916
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             2843..4471
                     /label="YES1(NM_005433)"
                     /note="YES1(NM_005433)"
                     /gene="YES1"
     promoter        4503..5002
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5023..5622
                     /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    5636..6224
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6431..7064
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7193..7209)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7217..7233
                     /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(7241..7271)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7286..7307
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7595..8183)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8354..9214)
                     /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(9215..9319)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9367..9501
                     /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 ctcaagcttc gaattcgcca ccatgggctg cattaaaagt aaagaaaaca aaagtccagc
     2881 cattaaatac agacctgaaa atactccaga gcctgtcagt acaagtgtga gccattatgg
     2941 agcagaaccc actacagtgt caccatgtcc gtcatcttca gcaaagggaa cagcagttaa
     3001 tttcagcagt ctttccatga caccatttgg aggatcctca ggggtaacgc cttttggagg
     3061 tgcatcttcc tcattttcag tggtgccaag ttcatatcct gctggtttaa caggtggtgt
     3121 tactatattt gtggccttat atgattatga agctagaact acagaagacc tttcatttaa
     3181 gaagggtgaa agatttcaaa taattaacaa tacggaagga gattggtggg aagcaagatc
     3241 aatcgctaca ggaaagaatg gttatatccc gagcaattat gtagcgcctg cagattccat
     3301 tcaggcagaa gaatggtatt ttggcaaaat ggggagaaaa gatgctgaaa gattactttt
     3361 gaatcctgga aatcaacgag gtattttctt agtaagagag agtgaaacaa ctaaaggtgc
     3421 ttattccctt tctattcgtg attgggatga gataaggggt gacaatgtga aacactacaa
     3481 aattaggaaa cttgacaatg gtggatacta tatcacaacc agagcacaat ttgatactct
     3541 gcagaaattg gtgaaacact acacagaaca tgctgatggt ttatgccaca agttgacaac
     3601 tgtgtgtcca actgtgaaac ctcagactca aggtctagca aaagatgctt gggaaatccc
     3661 tcgagaatct ttgcgactag aggttaaact aggacaagga tgtttcggcg aagtgtggat
     3721 gggaacatgg aatggaacca cgaaagtagc aatcaaaaca ctaaaaccag gtacaatgat
     3781 gccagaagct ttccttcaag aagctcagat aatgaaaaaa ttaagacatg ataaacttgt
     3841 tccactatat gctgttgttt ctgaagaacc aatttacatt gtcactgaat ttatgtcaaa
     3901 aggaagctta ttagatttcc ttaaggaagg agatggaaag tatttgaagc ttccacagct
     3961 ggttgatatg gctgctcaga ttgctgatgg tatggcatat attgaaagaa tgaactatat
     4021 tcaccgagat cttcgggctg ctaatattct tgtaggagaa aatcttgtgt gcaaaatagc
     4081 agactttggt ttagcaaggt taattgaaga caatgaatac acagcaagac aaggtgcaaa
     4141 atttccaatc aaatggacag ctcctgaagc tgcactgtat ggtcggttta caataaagtc
     4201 tgatgtctgg tcatttggaa ttctgcaaac agaactagta acaaagggcc gagtgccata
     4261 tccaggtatg gtgaaccgtg aagtattaga acaagtggag cgaggataca ggatgccgtg
     4321 ccctcagggc tgtccagaat ccctccatga attgatgaat ctgtgttgga agaaggaccc
     4381 tgatgaaaga ccaacatttg aatatattca gtccttcttg gaagactact tcactgctac
     4441 agagccacag taccagccag gagaaaattt ataaggatcc cgcgactcta gataattcta
     4501 ccgggtaggg gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct
     4561 gggcacttgg cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag
     4621 gcgccaaccg gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt
     4681 caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc
     4741 acgtctcact agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct
     4801 ttggggcagc ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag
     4861 gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc
     4921 ctccggaggc ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc
     4981 ttcctcatct ccgggccttt cgacctgcag cccaagctta ccatgaccga gtacaagccc
     5041 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     5101 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     5161 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     5221 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     5281 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     5341 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     5401 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     5461 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     5521 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     5581 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaccgcgtct ggaacaatca
     5641 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     5701 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     5761 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     5821 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     5881 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     5941 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     6001 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     6061 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     6121 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     6181 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa ttaattctgc
     6241 agtcgagacc tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct
     6301 gattgtgcct ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag
     6361 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     6421 aagaggggac tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg
     6481 atctaccaca cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc
     6541 agatatccac tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta
     6601 gaagaggcca ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg
     6661 gatgacccgg agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac
     6721 gtggcccgag agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag
     6781 ggactttccg ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga
     6841 gccctcagat cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga
     6901 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     6961 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     7021 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca
     7081 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggccttg
     7141 acattgctag cgttttaccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     7201 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     7261 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     7321 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     7381 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     7441 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     7501 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     7561 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     7621 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     7681 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     7741 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     7801 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     7861 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     7921 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     7981 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     8041 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     8101 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     8161 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     8221 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     8281 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     8341 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     8401 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     8461 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     8521 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     8581 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     8641 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     8701 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     8761 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     8821 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     8881 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     8941 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     9001 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     9061 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     9121 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     9181 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     9241 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     9301 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     9361 gagatcaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     9421 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     9481 gtatcttatc atgtctggat caactggata actcaagcta accaaaatca tcccaaactt
     9541 cccaccccat accctattac cactgccaat tacctgtggt ttcatttact ctaaacctgt
     9601 gattcctctg aattattttc attttaaaga aattgtattt gttaaatatg tactacaaac
     9661 ttagtagttt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagt
//
