LOCUS       V022694                 9533 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022694
VERSION     V022694
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..4289
                     /label="Hnrnpk(NM_057141)"
                     /note="Hnrnpk(NM_057141)"
                     /gene="Hnrnpk"
     promoter        4321..4820
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4841..5440
                     /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    5454..6042
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6249..6882
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7011..7027)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7035..7051
                     /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(7059..7089)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7104..7125
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7413..8001)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8172..9032)
                     /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(9033..9137)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9185..9319
                     /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 atggagaccg aacagccaga agaaaccttc cctaacaccg
     2941 aaaccaatgg tgaatttggt aaacgccctg cagaagatat ggaagaggaa caagccttta
     3001 aaagatctag aaatactgat gagatggttg aattgcgcat tttgcttcag agcaagaatg
     3061 ctggggcagt gattggaaaa ggaggcaaga atattaaggc tctccgtaca gactacaatg
     3121 ccagtgtttc agtcccagac agcagtggcc ccgagcgcat actgagtatc agtgctgata
     3181 ttgagacgat tggagaaatt ctgaagaaaa tcatccctac tttggaagag ggcctgcagt
     3241 tgccatcacc cactgcaacc agccagctcc cgctcgaatc tgatgctgtg gaatgcttaa
     3301 attaccagca ttataaaggg agtgacttcg actgcgagct gagactgttg attcatcaaa
     3361 gtctggccgg aggaattatt ggtgttaaag gtgctaaaat caaagaactt cgagaaaaca
     3421 ctcagacaac aatcaagctt ttccaggaat gctgccctca ttctactgac agagttgttc
     3481 ttattggagg aaagcccgat agggttgtag aatgcatcaa gatcatcctt gaccttatat
     3541 ctgagtctcc catcaaagga cgtgcacaac cttatgatcc caacttctat gatgaaacct
     3601 atgattatgg tggttttaca atgatgtttg atgaccgccg aggacgacct gtgggattcc
     3661 ccatgcgggg aagaggtggt tttgacagaa tgcctcctgg tcggggtggg cgtcccatgc
     3721 ctccttctag aagagattat gatgatatga gccctcgtcg aggaccacca ccaccaccac
     3781 ctggtcgagg tggacggggt ggcagcagag ctcggaatct tcctcttcct ccaccaccac
     3841 cacccagagg gggagatctc atggcttacg acagaagagg acggcctgga gaccgctatg
     3901 atggcatggt tgggttcagt gctgatgaaa cttgggactc tgcaattgac acatggagcc
     3961 catcagaatg gcaaatggct tatgaaccac agggtggttc tggatatgat tattcttatg
     4021 cagggggccg tggctcgtat ggtgatcttg gcggacctat tatcactaca caagtaacta
     4081 ttcccaaaga tttggctgga tctattattg gcaaaggtgg tcagcggatt aaacaaattc
     4141 gtcatgagtc tggagcatct atcaaaattg atgaaccttt agaaggatct gaagatcgga
     4201 tcattaccat tacgggaaca caggaccaga tacagaacgc acagtatttg ctgcagaaca
     4261 gtgtgaagca gtattctgga aagtttttct aaggatcccg cgactctaga taattctacc
     4321 gggtagggga ggcgcttttc ccaaggcagt ctggagcatg cgctttagca gccccgctgg
     4381 gcacttggcg ctacacaagt ggcctctggc ctcgcacaca ttccacatcc accggtaggc
     4441 gccaaccggc tccgttcttt ggtggcccct tcgcgccacc ttctactcct cccctagtca
     4501 ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac
     4561 gtctcactag tctcgtgcag atggacagca ccgctgagca atggaagcgg gtaggccttt
     4621 ggggcagcgg ccaatagcag ctttgctcct tcgctttctg ggctcagagg ctgggaaggg
     4681 gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct
     4741 ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt
     4801 cctcatctcc gggcctttcg acctgcagcc caagcttacc atgaccgagt acaagcccac
     4861 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt
     4921 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac
     4981 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     5041 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     5101 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     5161 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     5221 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     5281 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     5341 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     5401 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga ccgcgtctgg aacaatcaac
     5461 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     5521 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     5581 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     5641 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     5701 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     5761 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     5821 ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg
     5881 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     5941 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     6001 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaatt aattctgcag
     6061 tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga
     6121 ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt
     6181 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     6241 gaggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat
     6301 ctaccacaca caaggctact tccctgatta gcagaactac acaccagggc caggggtcag
     6361 atatccactg acctttggat ggtgctacaa gctagtacca gttgagccag ataaggtaga
     6421 agaggccaat aaaggagaga acaccagctt gttacaccct gtgagcctgc atgggatgga
     6481 tgacccggag agagaagtgt tagagtggag gtttgacagc cgcctagcat ttcatcacgt
     6541 ggcccgagag ctgcatccgg agtacttcaa gaactgctga tatcgagctt gctacaaggg
     6601 actttccgct ggggactttc cagggaggcg tggcctgggc gggactgggg agtggcgagc
     6661 cctcagatcc tgcatataag cagctgcttt ttgcctgtac tgggtctctc tggttagacc
     6721 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     6781 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     6841 gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc
     6901 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggccttgac
     6961 attgctagcg ttttaccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     7021 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     7081 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     7141 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     7201 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     7261 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     7321 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     7381 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     7441 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     7501 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     7561 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     7621 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     7681 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     7741 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     7801 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     7861 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     7921 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     7981 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     8041 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     8101 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     8161 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     8221 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     8281 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     8341 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     8401 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     8461 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     8521 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     8581 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     8641 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     8701 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     8761 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     8821 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     8881 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     8941 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     9001 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     9061 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     9121 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     9181 gatcaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     9241 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     9301 atcttatcat gtctggatca actggataac tcaagctaac caaaatcatc ccaaacttcc
     9361 caccccatac cctattacca ctgccaatta cctgtggttt catttactct aaacctgtga
     9421 ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt taaatatgta ctacaaactt
     9481 agtagttttt aaagaaattg tatttgttaa atatgtacta caaacttagt agt
//
