LOCUS       V021226                 9957 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021226
VERSION     V021226
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             2807..3514
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
                     /label="mCherry"
     CDS             3530..3556
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3557..4693
                     /label="MAPK3(NM_002746)"
                     /note="MAPK3(NM_002746)"
                     /gene="MAPK3"
     CDS             4703..4750
                     /codon_start=1
                     /product="bipartite nuclear localization signal from
                     nucleoplasmin"
                     /transl_table=1
                     /translation="KRPAATKKAGQAKKKK"
                     /label="nucleoplasmin NLS"
     promoter        4792..5291
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5312..5911
                     /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    5925..6513
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6720..7353
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7482..7498)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7506..7522
                     /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(7530..7560)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7575..7596
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7884..8472)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8643..9503)
                     /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(9504..9608)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9656..9790
                     /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 tctaccggtc gccaccatgg tgagcaaggg
     2821 cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg
     2881 ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
     2941 cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat
     3001 cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc
     3061 cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
     3121 ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta
     3181 caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac
     3241 catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga
     3301 gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac
     3361 ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga
     3421 catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg
     3481 ccactccacc ggcggcatgg acgagctgta caagggagga gggggaagct acccatacga
     3541 cgtaccagat tacgctatgg cggcggcggc ggctcagggg ggcgggggcg gggagccccg
     3601 tagaaccgag ggggtcggcc cgggggtccc gggggaggtg gagatggtga aggggcagcc
     3661 gttcgacgtg ggcccgcgct acacgcagtt gcagtacatc ggcgagggcg cgtacggcat
     3721 ggtcagctcg gcctatgacc acgtgcgcaa gactcgcgtg gccatcaaga agatcagccc
     3781 cttcgaacat cagacctact gccagcgcac gctccgggag atccagatcc tgctgcgctt
     3841 ccgccatgag aatgtcatcg gcatccgaga cattctgcgg gcgtccaccc tggaagccat
     3901 gagagatgtc tacattgtgc aggacctgat ggagactgac ctgtacaagt tgctgaaaag
     3961 ccagcagctg agcaatgacc atatctgcta cttcctctac cagatcctgc ggggcctcaa
     4021 gtacatccac tccgccaacg tgctccaccg agatctaaag ccctccaacc tgctcatcaa
     4081 caccacctgc gaccttaaga tttgtgattt cggcctggcc cggattgccg atcctgagca
     4141 tgaccacacc ggcttcctga cggagtatgt ggctacgcgc tggtaccggg ccccagagat
     4201 catgctgaac tccaagggct ataccaagtc catcgacatc tggtctgtgg gctgcattct
     4261 ggctgagatg ctctctaacc ggcccatctt ccctggcaag cactacctgg atcagctcaa
     4321 ccacattctg ggcatcctgg gctccccatc ccaggaggac ctgaattgta tcatcaacat
     4381 gaaggcccga aactacctac agtctctgcc ctccaagacc aaggtggctt gggccaagct
     4441 tttccccaag tcagactcca aagcccttga cctgctggac cggatgttaa cctttaaccc
     4501 caataaacgg atcacagtgg aggaagcgct ggctcacccc tacctggagc agtactatga
     4561 cccgacggat gagccagtgg ccgaggagcc cttcaccttc gccatggagc tggatgacct
     4621 acctaaggag cggctgaagg agctcatctt ccaggagaca gcacgcttcc agcccggagt
     4681 gctggaggcc cccactcgag ctaagcgacc tgccgccaca aagaaggctg gacaggctaa
     4741 gaagaagaaa tagggatcca ccggatctag ataactgatc ataattctac cgggtagggg
     4801 aggcgctttt cccaaggcag tctggagcat gcgctttagc agccccgctg ggcacttggc
     4861 gctacacaag tggcctctgg cctcgcacac attccacatc caccggtagg cgccaaccgg
     4921 ctccgttctt tggtggcccc ttcgcgccac cttctactcc tcccctagtc aggaagttcc
     4981 cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta
     5041 gtctcgtgca gatggacagc accgctgagc aatggaagcg ggtaggcctt tggggcagcg
     5101 gccaatagca gctttgctcc ttcgctttct gggctcagag gctgggaagg ggtgggtccg
     5161 ggggcgggct caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc
     5221 cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct tcctcatctc
     5281 cgggcctttc gacctgcagc ccaagcttac catgaccgag tacaagccca cggtgcgcct
     5341 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta
     5401 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca
     5461 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     5521 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     5581 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     5641 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     5701 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     5761 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
     5821 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     5881 ctggtgcatg acccgcaagc ccggtgcctg accgcgtctg gaacaatcaa cctctggatt
     5941 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     6001 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     6061 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     6121 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     6181 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     6241 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     6301 ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct
     6361 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     6421 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     6481 cgagtcggat ctccctttgg gccgcctccc cgcctggaat taattctgca gtcgagacct
     6541 agaaaaacat ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg
     6601 gctagaagca caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag
     6661 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa agaggggact
     6721 ggaagggcta attcactccc aacgaagaca agatatcctt gatctgtgga tctaccacac
     6781 acaaggctac ttccctgatt agcagaacta cacaccaggg ccaggggtca gatatccact
     6841 gacctttgga tggtgctaca agctagtacc agttgagcca gataaggtag aagaggccaa
     6901 taaaggagag aacaccagct tgttacaccc tgtgagcctg catgggatgg atgacccgga
     6961 gagagaagtg ttagagtgga ggtttgacag ccgcctagca tttcatcacg tggcccgaga
     7021 gctgcatccg gagtacttca agaactgctg atatcgagct tgctacaagg gactttccgc
     7081 tggggacttt ccagggaggc gtggcctggg cgggactggg gagtggcgag ccctcagatc
     7141 ctgcatataa gcagctgctt tttgcctgta ctgggtctct ctggttagac cagatctgag
     7201 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     7261 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     7321 gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc
     7381 agtatttata acttgcaaag aaatgaatat cagagagtga gaggccttga cattgctagc
     7441 gttttaccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     7501 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     7561 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     7621 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     7681 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     7741 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     7801 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     7861 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     7921 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     7981 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     8041 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     8101 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     8161 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     8221 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     8281 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     8341 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     8401 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     8461 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     8521 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     8581 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     8641 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     8701 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     8761 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     8821 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     8881 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     8941 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     9001 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     9061 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     9121 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     9181 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     9241 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     9301 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     9361 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     9421 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     9481 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     9541 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     9601 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcaactt
     9661 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     9721 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     9781 tgtctggatc aactggataa ctcaagctaa ccaaaatcat cccaaacttc ccaccccata
     9841 ccctattacc actgccaatt acctgtggtt tcatttactc taaacctgtg attcctctga
     9901 attattttca ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagt
//
