LOCUS       V045881                 9748 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045881
VERSION     V045881
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      2809..2818
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             2818..2883
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             2890..4440
                     /label="ZNF582(NM_001320371)"
                     /note="ZNF582(NM_001320371)"
                     /gene="ZNF582"
     misc_feature    4451..5037
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES2"
     CDS             5057..5656
                     /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    5670..6258
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6465..7098
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7226..7242)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7250..7266
                     /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(7274..7304)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7319..7340
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7628..8216)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8387..9247)
                     /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(9248..9352)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9400..9534
                     /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 tctatttccg gtgaattcgc caccatggac
     2821 tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac
     2881 aagctcgaga tgtcccttgg gtcagaattg ttcagggatg tggccatagt cttctcccaa
     2941 gaagaatggc agtggttggc acctgctcag agggatttgt acagagacgt gatgttggag
     3001 acctacagca acctggtctc actgggtctt gccgtttcca aacctgatgt gatctccttc
     3061 ctagagcaag gcaaagagcc ctggatggtg gagagagtgg tgtctggagg cctgtgtcca
     3121 gtattggagt ccagatatga taccaaggaa ttatttccaa agcagcatgt ttatgaagtc
     3181 gaatcacccc aatgggagat aatggaaagc cttacaagtt atggtcttga gtgttcaagt
     3241 ttccaagatg attgggaatg cagaaaccag tttgacagac aacagggaaa tccagacaga
     3301 catttccatc agatgatcat cagacatgaa gaaatgccca cttttgacca gcatgcatcc
     3361 cttacttttt atcagaaaat tcatactaga gaaaaacctt ttgggtataa taaatgtaga
     3421 aaagacttct ggcaaaagga actccttatt aatcatcaag gaatttatac taatgagaaa
     3481 ccctataaat gtaaggaatg tgggaaggcc tttaaatatg gctcacgact aattcaacat
     3541 gagaatattc attctggtaa gaaaccatat gaatgtaagg aatgtggaaa ggccttcaat
     3601 tctggttcaa attttataca acatcagaga gttcatactg gtgagaaacc gtatgaatgt
     3661 aaagattgtg agaaggcctt tagtcgaagc tcacagttga ttgaacatca gcgaactcac
     3721 acaggcgaga aaccctatca gtgtaaggaa tgtggcaagg ccttcaatcg gatctcacat
     3781 cttaaagtac attatagaat tcatactggt gaaaaaccct atgcatgcaa ggaatgtggg
     3841 aaaaccttta gtcatcgttc tcagttgatt caacatcaga ctgttcatac tggcaggaaa
     3901 ctctatgaat gtaaggaatg tgggaaggct tttaatcaag gctcaactct tatacgacat
     3961 cagagaattc ataccggtga gaaaccctat gagtgcaaag tatgtggaaa ggcctttaga
     4021 gtgagctcac aactcaagca acatcagaga attcacactg gagagaaacc ctaccaatgt
     4081 aaggtatgtg gtagggcctt caaacgggtc tcacatctta ctgtacatta cagaattcat
     4141 acaggtgaaa agccatatga atgtaaggaa tgtgggaagg cttttagtca ttgctcacaa
     4201 ttgattcatc atcaggttat tcacactgag aaaaagccct atgaatataa ggaatgtgag
     4261 aagaccttga gtcatgattc aactaccgtt caacctcaga gaatgcataa tagagaaaca
     4321 catgtgaata taataaatgt agaaaagcct tccatcagca gttacccctt actaatcatc
     4381 agagaattta tgttggcaag caaccatatg aatggaagta atggggaaag cccattagcc
     4441 taggatcccg cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat
     4501 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
     4561 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
     4621 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
     4681 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
     4741 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
     4801 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
     4861 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
     4921 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
     4981 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataagcttgc cacaacccac
     5041 aaggagacga ccttccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     5101 cgtcccccgg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     5161 caccgtcgac ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     5221 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     5281 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     5341 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     5401 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     5461 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     5521 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     5581 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     5641 caagcccggt gcctagacgc gtctggaaca atcaacctct ggattacaaa atttgtgaaa
     5701 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     5761 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     5821 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     5881 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     5941 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     6001 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     6061 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     6121 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     6181 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     6241 tttgggccgc ctccccgcct ggaattaatt ctgcagtcga gacctagaaa aacatggagc
     6301 aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag aagcacaaga
     6361 ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa tgacttacaa
     6421 ggcagctgta gatcttagcc actttttaaa agaaaagagg ggactggaag ggctaattca
     6481 ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag gctacttccc
     6541 tgattagcag aactacacac cagggccagg ggtcagatat ccactgacct ttggatggtg
     6601 ctacaagcta gtaccagttg agccagataa ggtagaagag gccaataaag gagagaacac
     6661 cagcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag aagtgttaga
     6721 gtggaggttt gacagccgcc tagcatttca tcacgtggcc cgagagctgc atccggagta
     6781 cttcaagaac tgctgatatc gagcttgcta caagggactt tccgctgggg actttccagg
     6841 gaggcgtggc ctgggcggga ctggggagtg gcgagccctc agatcctgca tataagcagc
     6901 tgctttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     6961 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     7021 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     7081 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
     7141 caaagaaatg aatatcagag agtgagaggc cttgacattg ctagcgttta ccgtcgacct
     7201 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     7261 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     7321 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     7381 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     7441 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     7501 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     7561 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     7621 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     7681 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     7741 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     7801 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     7861 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     7921 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     7981 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     8041 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     8101 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     8161 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     8221 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     8281 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     8341 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     8401 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     8461 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     8521 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     8581 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     8641 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     8701 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     8761 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     8821 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     8881 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     8941 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     9001 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     9061 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     9121 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     9181 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     9241 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     9301 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     9361 cccgaaaagt gccacctgac gtcgacggat cgggagatca acttgtttat tgcagcttat
     9421 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     9481 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcaactgg
     9541 ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc cataccctat taccactgcc
     9601 aattacctgt ggtttcattt actctaaacc tgtgattcct ctgaattatt ttcattttaa
     9661 agaaattgta tttgttaaat atgtactaca aacttagtag tttttaaaga aattgtattt
     9721 gttaaatatg tactacaaac ttagtagt
//
