LOCUS       V032019                10361 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032019
VERSION     V032019
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"
     regulatory      2820..2829
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             2829..2894
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             2901..4520
                     /label="GPR75(NM_006794)"
                     /note="GPR75(NM_006794)"
                     /gene="GPR75"
     promoter        5149..5648
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     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(7839..7855)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7863..7879
                     /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(7887..7917)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7932..7953
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8241..8829)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9000..9860)
                     /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(9861..9965)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    10013..10147
                     /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 ccaccatgga ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg
     2881 atgacgatga caaggaattc atgaactcaa caggccacct tcaggatgcc cccaatgcca
     2941 cctcgctcca tgtgcctcac tcacaggaag gaaacagcac ctctctccag gagggtcttc
     3001 aggatctcat ccacacagcc accttggtga cctgtacttt tctactggcg gtcatcttct
     3061 gcctgggttc ctatggcaac ttcattgtct tcttgtcctt cttcgatcca gccttcagga
     3121 aattcagaac caactttgat ttcatgatcc tgaacctgtc cttctgtgac ctcttcattt
     3181 gtggagtgac agcccccatg ttcacctttg tgttattctt cagctcagcc agtagtatcc
     3241 cggatgcttt ctgcttcact ttccatctca ccagttcagg cttcatcatc atgtctctga
     3301 agacagtggc agtgatcgcc ctgcaccggc tccggatggt gttggggaaa cagcctaatc
     3361 gcacggcctc ctttccctgc accgtactcc tcaccctgct tctctgggcc accagtttca
     3421 cccttgccac cttggctacc ttgaaaacca gcaagtccca cctctgtctt cccatgtcca
     3481 gtctgattgc tggaaaaggg aaagccattt tgtctctcta tgtggtcgac ttcaccttct
     3541 gtgttgctgt ggtctctgtc tcttacatca tgattgctca gaccctgcgg aagaacgctc
     3601 aagtcagaaa gtgcccccct gtaatcacag tcgatgcttc cagaccacag cctttcatgg
     3661 gggtccctgt gcagggaggt ggagatccca tccagtgtgc catgccggct ctgtatagga
     3721 accagaatta caacaaactg cagcacgttc agacccgtgg atataccaag agtcccaacc
     3781 aactggtcac ccctgcagca agccgactcc agctcgtatc agccatcaac ctctccactg
     3841 ccaaggattc caaagccgtg gtcacctgtg tgatcattgt gctgtcagtc ctggtgtgct
     3901 gtcttccact ggggatttcc ttggtacagg tggttctctc cagcaatggg agcttcattc
     3961 tttaccagtt tgaattgttt ggatttactc ttatattttt caagtcagga ttaaaccctt
     4021 ttatatattc tcggaacagt gcagggctga gaaggaaagt gctctggtgc ctccaataca
     4081 taggcctggg ttttttctgc tgcaaacaaa agactcgact tcgagccatg ggaaaaggga
     4141 acctcgaagt caacagaaac aaatcctccc atcatgaaac aaactctgcc tacatgttat
     4201 ctccaaagcc acagaagaaa tttgtggacc aggcttgtgg cccaagtcat tcaaaagaaa
     4261 gtatggtgag tcccaagatc tctgctggac atcaacactg tggtcagagc agctcgaccc
     4321 ccatcaacac tcggattgaa ccttactaca gcatctataa cagcagccct tcccaggagg
     4381 agagcagccc atgtaactta cagccagtaa actcttttgg atttgccaat tcatatattg
     4441 ccatgcatta tcacaccact aatgacttag tgcaggaata tgacagcact tcagccaagc
     4501 agattccagt cccctccgtt accagctctg gcagcagcgg cggcggcggc agcggcggcg
     4561 gcggcagcag cggcatggtc ttcacactcg aagatttcgt tggggactgg gaacagacag
     4621 ccgcctacaa cctggaccaa gtccttgaac agggaggtgt gtccagtttg ctgcagaatc
     4681 tcgccgtgtc cgtaactccg atccaaagga ttgtccggag cggtgaaaat gccctgaaga
     4741 tcgacatcca tgtcatcatc ccgtatgaag gtctgagcgc cgaccaaatg gcccagatcg
     4801 aagaggtgtt taaggtggtg taccctgtgg atgatcatca ctttaaggtg atcctgccct
     4861 atggcacact ggtaatcgac ggggttacgc cgaacatgct gaactatttc ggacggccgt
     4921 atgaaggcat cgccgtgttc gacggcaaaa agatcactgt aacagggacc ctgtggaacg
     4981 gcaacaaaat tatcgacgag cgcctgatca cccccgacgg ctccatgctg ttccgagtaa
     5041 ccatcaacag tgggagttcc ggtggtggcg ggagcggagg tggaggctcg agcggtggag
     5101 ctcaggggaa ttcagtctaa ggatcccgcg actctagata attctaccgg gtaggggagg
     5161 cgcttttccc aaggcagtct ggagcatgcg ctttagcagc cccgctgggc acttggcgct
     5221 acacaagtgg cctctggcct cgcacacatt ccacatccac cggtaggcgc caaccggctc
     5281 cgttctttgg tggccccttc gcgccacctt ctactcctcc cctagtcagg aagttccccc
     5341 ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga agtagcacgt ctcactagtc
     5401 tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt aggcctttgg ggcagcggcc
     5461 aatagcagct ttgctccttc gctttctggg ctcagaggct gggaaggggt gggtccgggg
     5521 gcgggctcag gggcgggctc aggggcgggg cgggcgcccg aaggtcctcc ggaggcccgg
     5581 cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt ctcctcttcc tcatctccgg
     5641 gcctttcgac ctgcagccca agcttaccat gaccgagtac aagcccacgg tgcgcctcgc
     5701 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     5761 cgccacgcgc cacaccgtcg atccggaccg 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 gtgcctgacc 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 ttaccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     7861 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     7921 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     7981 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     8041 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     8101 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     8161 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     8221 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     8281 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     8341 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     8401 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     8461 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     8521 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     8581 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     8641 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     8701 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     8761 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     8821 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     8881 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     8941 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     9001 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     9061 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     9121 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     9181 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     9241 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     9301 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     9361 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     9421 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     9481 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     9541 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     9601 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     9661 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     9721 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     9781 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     9841 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     9901 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     9961 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tcaacttgtt
    10021 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
    10081 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
    10141 ctggatcaac tggataactc aagctaacca aaatcatccc aaacttccca ccccataccc
    10201 tattaccact gccaattacc tgtggtttca tttactctaa acctgtgatt cctctgaatt
    10261 attttcattt taaagaaatt gtatttgtta aatatgtact acaaacttag tagtttttaa
    10321 agaaattgta tttgttaaat atgtactaca aacttagtag t
//
