LOCUS       V027864                 9841 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027864
VERSION     V027864
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..4537
                     /label="WSCD2(NM_014653)"
                     /note="WSCD2(NM_014653)"
                     /gene="WSCD2"
     CDS             4544..4609
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4629..5128
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5149..5748
                     /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    5762..6350
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6557..7190
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7319..7335)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7343..7359
                     /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(7367..7397)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7412..7433
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7721..8309)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8480..9340)
                     /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(9341..9445)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9493..9627
                     /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 ccatggccaa gctctggttc aaattccagc ggtacttccg
     2881 ccggaaacct gtgcgcttct ttaccttcct ggcactctac ctgactgctg ggagccttgt
     2941 cttccttcac tctggctttg tgggccagcc cgctgtctcg gggaaccagg cgaaccccgc
     3001 tgctgcagga ggcccagctg agggtgctga gctgtccttc ttgggtgaca tgcatctggg
     3061 cagaggtttc cgggacacag gtgaagcctc aagcattgct cgcaggtacg gaccctggtt
     3121 caagggcaag gatgggaatg agagagccaa gcttggcgac tacggtggag cctggagccg
     3181 agccctcaag gggagggttg tccgggagaa ggaggaagag cgagccaagt acatcggctg
     3241 ctacctggat gacacccaga gtcgggccct tcgaggagtg tccttttttg actacaaaaa
     3301 gatgaccatc ttccgttgcc aggacaactg tgctgaacgg ggttacctgt atggcgggct
     3361 ggagttcggc gccgagtgct actgcggcca caagatccag gcgacgaacg tgagcgaggc
     3421 agagtgcgac atggagtgca agggcgagcg aggcagcgtg tgcggcggcg ccaaccgcct
     3481 ctctgtctac cggctgcagc tggcccagga gtcggcccgc aggtatggaa gtgcagtgtt
     3541 ccggggctgc ttccgcaggc ccgacaacct ttccctggcc ttacccgtga cagctgccat
     3601 gctgaacatg tctgtggaca aatgcgtgga cttctgcact gagaaggagt acccgctggc
     3661 agctcttgca ggcaccgcct gccactgtgg gtttcccacc acccgattcc cgctccatga
     3721 cagagaggat gagcagctct gtgcccagaa gtgcagcgcg gaggagtttg agagctgcgg
     3781 gactcctagt tacttcattg tgtaccagac acaagtccaa gacaaccgtt gcatggacag
     3841 aaggttcctg ccaggcaagt ccaagcagct cattgctttg gccagcttcc caggtgctgg
     3901 caacacgtgg gctcgccacc tcattgaatt ggccacaggc ttctacactg gcagctacta
     3961 cttcgatggc tccctctaca acaaagggtt taaaggtgag cgggaccact ggcgcagcgg
     4021 acggaccatc tgcatcaaga cgcacgaaag cggccagaaa gagatcgagg ccttcgacgc
     4081 cgccatcctg ctcatccgca acccctacaa agccctcatg gctgagttca accgcaagta
     4141 cggcggccac ataggctttg ctgcgcatgc ccactggaag ggcaaagagt ggccagagtt
     4201 cgtgaggaac tatgccccgt ggtgggccac tcacacactg gactggctca agtttggcaa
     4261 gaaggtgctg gtggtgcact ttgaggacct gaagcaggac ctctttgtcc agctgggccg
     4321 gatggtcagc ctgctgggcg tggctgtcag ggaggaccgg ctgctctgtg tggagagcca
     4381 gaaggatggc aacttcaagc gctcagggct ccggaagctc gagtatgacc cctatactgc
     4441 ggacatgcag aagaccatct ctgcctacat caagatggtg gatgcagccc tcaaagggcg
     4501 gaacctaacg ggtgtccccg atgactacta cccaagagga tccgactaca aagaccatga
     4561 cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt agtctagata
     4621 attctaccgg gtaggggagg cgcttttccc aaggcagtct ggagcatgcg ctttagcagc
     4681 cccgctgggc acttggcgct acacaagtgg cctctggcct cgcacacatt ccacatccac
     4741 cggtaggcgc caaccggctc cgttctttgg tggccccttc gcgccacctt ctactcctcc
     4801 cctagtcagg aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga
     4861 agtagcacgt ctcactagtc tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt
     4921 aggcctttgg ggcagcggcc aatagcagct ttgctccttc gctttctggg ctcagaggct
     4981 gggaaggggt gggtccgggg gcgggctcag gggcgggctc aggggcgggg cgggcgcccg
     5041 aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt
     5101 ctcctcttcc tcatctccgg gcctttcgac ctgcagccca agcttaccat gaccgagtac
     5161 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
     5221 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
     5281 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
     5341 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
     5401 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
     5461 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
     5521 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
     5581 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     5641 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
     5701 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacc gcgtctggaa
     5761 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     5821 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     5881 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     5941 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     6001 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     6061 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     6121 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     6181 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     6241 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     6301 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaattaa
     6361 ttctgcagtc gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc
     6421 aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca
     6481 cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     6541 aaagaaaaga ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat
     6601 ctgtggatct accacacaca aggctacttc cctgattagc agaactacac accagggcca
     6661 ggggtcagat atccactgac ctttggatgg tgctacaagc tagtaccagt tgagccagat
     6721 aaggtagaag aggccaataa aggagagaac accagcttgt tacaccctgt gagcctgcat
     6781 gggatggatg acccggagag agaagtgtta gagtggaggt ttgacagccg cctagcattt
     6841 catcacgtgg cccgagagct gcatccggag tacttcaaga actgctgata tcgagcttgc
     6901 tacaagggac tttccgctgg ggactttcca gggaggcgtg gcctgggcgg gactggggag
     6961 tggcgagccc tcagatcctg catataagca gctgcttttt gcctgtactg ggtctctctg
     7021 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     7081 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     7141 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc
     7201 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag
     7261 gccttgacat tgctagcgtt ttaccgtcga cctctagcta gagcttggcg taatcatggt
     7321 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     7381 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     7441 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     7501 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     7561 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     7621 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     7681 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     7741 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     7801 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     7861 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     7921 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     7981 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     8041 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     8101 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     8161 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     8221 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     8281 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     8341 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     8401 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     8461 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     8521 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     8581 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     8641 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     8701 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     8761 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     8821 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     8881 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     8941 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     9001 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     9061 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     9121 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     9181 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     9241 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     9301 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     9361 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     9421 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     9481 gatcgggaga tcaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     9541 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     9601 atcaatgtat cttatcatgt ctggatcaac tggataactc aagctaacca aaatcatccc
     9661 aaacttccca ccccataccc tattaccact gccaattacc tgtggtttca tttactctaa
     9721 acctgtgatt cctctgaatt attttcattt taaagaaatt gtatttgtta aatatgtact
     9781 acaaacttag tagtttttaa agaaattgta tttgttaaat atgtactaca aacttagtag
     9841 t
//
