LOCUS       V032017                10231 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032017
VERSION     V032017
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..4966
                     /label="MRE11(NM_005591)"
                     /note="MRE11(NM_005591)"
                     /gene="MRE11"
     CDS             4973..4999
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     promoter        5019..5518
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5539..6138
                     /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    6152..6740
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6947..7580
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7709..7725)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7733..7749
                     /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(7757..7787)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7802..7823
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8111..8699)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8870..9730)
                     /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(9731..9835)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9883..10017
                     /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 ccatgagtac tgcagatgca cttgatgatg aaaacacatt
     2881 taaaatatta gttgcaacag atattcatct tggatttatg gagaaagatg cagtcagagg
     2941 aaatgatacg tttgtaacac tcgatgaaat tttaagactt gcccaggaaa atgaagtgga
     3001 ttttattttg ttaggtggtg atctttttca tgaaaataag ccctcaagga aaacattaca
     3061 tacctgcctc gagttattaa gaaaatattg tatgggtgat cggcctgtcc agtttgaaat
     3121 tctcagtgat cagtcagtca actttggttt tagtaagttt ccatgggtga actatcaaga
     3181 tggcaacctc aacatttcaa ttccagtgtt tagtattcat ggcaatcatg acgatcccac
     3241 aggggcagat gcactttgtg ccttggacat tttaagttgt gctggatttg taaatcactt
     3301 tggacgttca atgtctgtgg agaagataga cattagtccg gttttgcttc aaaaaggaag
     3361 cacaaagatt gcgctatatg gtttaggatc cattccagat gaaaggctct atcgaatgtt
     3421 tgtcaataaa aaagtaacaa tgttgagacc aaaggaagat gagaactctt ggtttaactt
     3481 atttgtgatt catcagaaca ggagtaaaca tggaagtact aacttcattc cagaacaatt
     3541 tttggatgac ttcattgatc ttgttatctg gggccatgaa catgagtgta aaatagctcc
     3601 aaccaaaaat gaacaacagc tgttttatat ctcacaacct ggaagctcag tggttacttc
     3661 tctttcccca ggagaagctg taaagaaaca tgttggtttg ctgcgtatta aagggaggaa
     3721 gatgaatatg cataaaattc ctcttcacac agtgcggcag tttttcatgg aggatattgt
     3781 tctagctaat catccagaca tttttaaccc agataatcct aaagtaaccc aagccataca
     3841 aagcttctgt ttggagaaga ttgaagaaat gcttgaaaat gctgaacggg aacgtctggg
     3901 taattctcac cagccagaga agcctcttgt acgactgcga gtggactata gtggaggttt
     3961 tgaacctttc agtgttcttc gctttagcca gaaatttgtg gatcgggtag ctaatccaaa
     4021 agacattatc cattttttca ggcatagaga acaaaaggaa aaaacaggag aagagatcaa
     4081 ctttgggaaa cttatcacaa agccttcaga aggaacaact ttaagggtag aagatcttgt
     4141 aaaacagtac tttcaaaccg cagagaagaa tgtgcagctc tcactgctaa cagaaagagg
     4201 gatgggtgaa gcagtacaag aatttgtgga caaggaggag aaagatgcca ttgaggaatt
     4261 agtgaaatac cagttggaaa aaacacagcg atttcttaaa gaacgtcata ttgatgccct
     4321 cgaagacaaa atcgatgagg aggtacgtcg tttcagagaa accagacaaa aaaatactaa
     4381 tgaagaagat gatgaagtcc gtgaggctat gaccagggcc agagcactca gatctcagtc
     4441 agaggagtct gcttctgcct ttagtgctga tgaccttatg agtatagatt tagcagaaca
     4501 gatggctaat gactctgatg atagcatctc agcagcaacc aacaaaggaa gaggccgagg
     4561 aagaggtcga agaggtggaa gagggcagaa ttcagcatcg agaggagggt ctcaaagagg
     4621 aagagcagac actggtctgg agacttctac ccgtagcagg aactcaaaga ctgctgtgtc
     4681 agcatctaga aatatgtcta ttatagatgc ctttaaatct acaagacagc agccttcccg
     4741 aaatgtcact actaagaatt attcagaggt gattgaggta gatgaatcag atgtggaaga
     4801 agacattttt cctaccactt caaagacaga tcaaaggtgg tccagcacat catccagcaa
     4861 aatcatgtcc cagagtcaag tatcgaaagg ggttgatttt gaatcaagtg aggatgatga
     4921 tgatgatcct tttatgaaca ctagttcttt aagaagaaat agaagaggat cctacccata
     4981 cgacgtacca gattacgctt agtctagata attctaccgg gtaggggagg cgcttttccc
     5041 aaggcagtct ggagcatgcg ctttagcagc cccgctgggc acttggcgct acacaagtgg
     5101 cctctggcct cgcacacatt ccacatccac cggtaggcgc caaccggctc cgttctttgg
     5161 tggccccttc gcgccacctt ctactcctcc cctagtcagg aagttccccc ccgccccgca
     5221 gctcgcgtcg tgcaggacgt gacaaatgga agtagcacgt ctcactagtc tcgtgcagat
     5281 ggacagcacc gctgagcaat ggaagcgggt aggcctttgg ggcagcggcc aatagcagct
     5341 ttgctccttc gctttctggg ctcagaggct gggaaggggt gggtccgggg gcgggctcag
     5401 gggcgggctc aggggcgggg cgggcgcccg aaggtcctcc ggaggcccgg cattctgcac
     5461 gcttcaaaag cgcacgtctg ccgcgctgtt ctcctcttcc tcatctccgg gcctttcgac
     5521 ctgcagccca agcttaccat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac
     5581 gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc
     5641 cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc
     5701 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg
     5761 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc
     5821 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg
     5881 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac
     5941 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc
     6001 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc
     6061 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc
     6121 cgcaagcccg gtgcctgacc gcgtctggaa caatcaacct ctggattaca aaatttgtga
     6181 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     6241 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     6301 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     6361 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     6421 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     6481 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     6541 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     6601 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     6661 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     6721 cctttgggcc gcctccccgc ctggaattaa ttctgcagtc gagacctaga aaaacatgga
     6781 gcaatcacaa gtagcaatac agcagctacc aatgctgatt gtgcctggct agaagcacaa
     6841 gaggaggagg aggtgggttt tccagtcaca cctcaggtac ctttaagacc aatgacttac
     6901 aaggcagctg tagatcttag ccacttttta aaagaaaaga ggggactgga agggctaatt
     6961 cactcccaac gaagacaaga tatccttgat ctgtggatct accacacaca aggctacttc
     7021 cctgattagc agaactacac accagggcca ggggtcagat atccactgac ctttggatgg
     7081 tgctacaagc tagtaccagt tgagccagat aaggtagaag aggccaataa aggagagaac
     7141 accagcttgt tacaccctgt gagcctgcat gggatggatg acccggagag agaagtgtta
     7201 gagtggaggt ttgacagccg cctagcattt catcacgtgg cccgagagct gcatccggag
     7261 tacttcaaga actgctgata tcgagcttgc tacaagggac tttccgctgg ggactttcca
     7321 gggaggcgtg gcctgggcgg gactggggag tggcgagccc tcagatcctg catataagca
     7381 gctgcttttt gcctgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     7441 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     7501 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     7561 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact
     7621 tgcaaagaaa tgaatatcag agagtgagag gccttgacat tgctagcgtt ttaccgtcga
     7681 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     7741 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     7801 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     7861 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     7921 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     7981 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     8041 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     8101 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     8161 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     8221 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     8281 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     8341 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     8401 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     8461 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     8521 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     8581 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     8641 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     8701 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     8761 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     8821 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     8881 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     8941 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     9001 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     9061 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     9121 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     9181 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     9241 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     9301 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     9361 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     9421 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     9481 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     9541 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     9601 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     9661 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     9721 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     9781 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     9841 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tcaacttgtt tattgcagct
     9901 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     9961 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcaac
    10021 tggataactc aagctaacca aaatcatccc aaacttccca ccccataccc tattaccact
    10081 gccaattacc tgtggtttca tttactctaa acctgtgatt cctctgaatt attttcattt
    10141 taaagaaatt gtatttgtta aatatgtact acaaacttag tagtttttaa agaaattgta
    10201 tttgttaaat atgtactaca aacttagtag t
//
