LOCUS       V039497                 9528 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039497
VERSION     V039497
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             2827..4254
                     /label="EGR2(NM_000399)"
                     /note="EGR2(NM_000399)"
                     /gene="EGR2"
     CDS             4261..4284
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     promoter        4316..4815
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4836..5435
                     /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    5449..6037
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6244..6877
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7006..7022)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7030..7046
                     /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(7054..7084)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7099..7120
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7408..7996)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8167..9027)
                     /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(9028..9132)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9180..9314
                     /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 gccaccatga tgaccgccaa ggccgtagac aaaatcccag taactctcag tggttttgtg
     2881 caccagctgt ctgacaacat ctacccggtg gaggacctcg ccgccacgtc ggtgaccatc
     2941 tttcccaatg ccgaactggg aggccccttt gaccagatga acggagtggc cggagatggc
     3001 atgatcaaca ttgacatgac tggagagaag aggtcgttgg atctcccata tcccagcagc
     3061 tttgctcccg tctctgcacc tagaaaccag accttcactt acatgggcaa gttctccatt
     3121 gaccctcagt accctggtgc cagctgctac ccagaaggca taatcaatat tgtgagtgca
     3181 ggcatcttgc aaggggtcac ttccccagct tcaaccacag cctcatccag cgtcacctct
     3241 gcctccccca acccactggc cacaggaccc ctgggtgtgt gcaccatgtc ccagacccag
     3301 cctgacctgg accacctgta ctctccgcca ccgcctcctc ctccttattc tggctgtgca
     3361 ggagacctct accaggaccc ttctgcgttc ctgtcagcag ccaccacctc cacctcttcc
     3421 tctctggcct acccaccacc tccttcctat ccatccccca agccagccac ggacccaggt
     3481 ctcttcccaa tgatcccaga ctatcctgga ttctttccat ctcagtgcca gagagaccta
     3541 catggtacag ctggcccaga ccgtaagccc tttccctgcc cactggacac cctgcgggtg
     3601 ccccctccac tcactccact ctctacaatc cgtaacttta ccctgggggg ccccagtgct
     3661 ggggtgaccg gaccaggggc cagtggaggc agcgagggac cccggctgcc tggtagcagc
     3721 tcagcagcag cagcagccgc cgccgccgcc gcctataacc cacaccacct gccactgcgg
     3781 cccattctga ggcctcgcaa gtaccccaac agacccagca agacgccggt gcacgagagg
     3841 ccctacccgt gcccagcaga aggctgcgac cggcggttct cccgctctga cgagctgaca
     3901 cggcacatcc gaatccacac tgggcataag cccttccagt gtcggatctg catgcgcaac
     3961 ttcagccgca gtgaccacct caccacccat atccgcaccc acaccggtga gaagcccttc
     4021 gcctgtgact actgtggccg aaagtttgcc cggagtgatg agaggaagcg ccacaccaag
     4081 atccacctga gacagaaaga gcggaaaagc agtgccccct ctgcatcggt gccagccccc
     4141 tctacagcct cctgctctgg gggcgtgcag cctgggggta ccctgtgcag cagtaacagc
     4201 agcagtcttg gcggagggcc gctcgcccct tgctcctctc ggacccggac acctgaattc
     4261 gattacaagg atgacgacga taagtgagga tcccgcgact ctagataatt ctaccgggta
     4321 ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt tagcagcccc gctgggcact
     4381 tggcgctaca caagtggcct ctggcctcgc acacattcca catccaccgg taggcgccaa
     4441 ccggctccgt tctttggtgg ccccttcgcg ccaccttcta ctcctcccct agtcaggaag
     4501 ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc
     4561 actagtctcg tgcagatgga cagcaccgct gagcaatgga agcgggtagg cctttggggc
     4621 agcggccaat agcagctttg ctccttcgct ttctgggctc agaggctggg aaggggtggg
     4681 tccgggggcg ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga
     4741 ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca
     4801 tctccgggcc tttcgacctg cagcccaagc ttaccatgac cgagtacaag cccacggtgc
     4861 gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg
     4921 actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc
     4981 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
     5041 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
     5101 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
     5161 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     5221 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     5281 aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
     5341 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     5401 gcacctggtg catgacccgc aagcccggtg cctgaccgcg tctggaacaa tcaacctctg
     5461 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     5521 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     5581 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     5641 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     5701 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     5761 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     5821 aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc
     5881 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     5941 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     6001 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaattaattc tgcagtcgag
     6061 acctagaaaa acatggagca atcacaagta gcaatacagc agctaccaat gctgattgtg
     6121 cctggctaga agcacaagag gaggaggagg tgggttttcc agtcacacct caggtacctt
     6181 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaagaggg
     6241 gactggaagg gctaattcac tcccaacgaa gacaagatat ccttgatctg tggatctacc
     6301 acacacaagg ctacttccct gattagcaga actacacacc agggccaggg gtcagatatc
     6361 cactgacctt tggatggtgc tacaagctag taccagttga gccagataag gtagaagagg
     6421 ccaataaagg agagaacacc agcttgttac accctgtgag cctgcatggg atggatgacc
     6481 cggagagaga agtgttagag tggaggtttg acagccgcct agcatttcat cacgtggccc
     6541 gagagctgca tccggagtac ttcaagaact gctgatatcg agcttgctac aagggacttt
     6601 ccgctgggga ctttccaggg aggcgtggcc tgggcgggac tggggagtgg cgagccctca
     6661 gatcctgcat ataagcagct gctttttgcc tgtactgggt ctctctggtt agaccagatc
     6721 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     6781 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     6841 ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt
     6901 attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggcc ttgacattgc
     6961 tagcgtttta ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     7021 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     7081 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     7141 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     7201 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     7261 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     7321 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     7381 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     7441 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     7501 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     7561 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     7621 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     7681 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     7741 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7801 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     7861 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     7921 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     7981 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     8041 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     8101 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     8161 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     8221 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     8281 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     8341 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     8401 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     8461 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     8521 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     8581 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     8641 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     8701 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     8761 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     8821 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     8881 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     8941 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     9001 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     9061 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     9121 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatca
     9181 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     9241 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     9301 atcatgtctg gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc
     9361 cataccctat taccactgcc aattacctgt ggtttcattt actctaaacc tgtgattcct
     9421 ctgaattatt ttcattttaa agaaattgta tttgttaaat atgtactaca aacttagtag
     9481 tttttaaaga aattgtattt gttaaatatg tactacaaac ttagtagt
//
