LOCUS       V022116                 9310 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022116
VERSION     V022116
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             2826..4049
                     /label="Map2k4(NM_001316367)"
                     /note="Map2k4(NM_001316367)"
                     /gene="Map2k4"
     CDS             4050..4067
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     promoter        4098..4597
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4618..5217
                     /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    5231..5819
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6026..6659
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6788..6804)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6812..6828
                     /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(6836..6866)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6881..6902
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7190..7778)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7949..8809)
                     /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(8810..8914)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    8962..9096
                     /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 ccaccatggc ggctccgagc ccgagcggcg gcggcggctc cgggggcggt ggcggcaccc
     2881 cggggcccat cggacctcca gcttctggcc acccggcggt cagcagcatg cagggctttc
     2941 agataaactt ctgtgaaaag gcacaaagta agcgcaaagc actgaagttg aattttgcaa
     3001 atccacctgt caaatcgaca gcacggttta ctctgaatcc taatactaca ggagtccaga
     3061 acccacacat agagagactg agaacacaca gcattgagtc atcaggaaaa ctgaagatct
     3121 cccctgaaca acactgggat ttcactgcag aggacttgaa agaccttgga gaaattggac
     3181 gaggagctta tggttctgtc aacaaaatgg tccacaaacc aagtgggcag ataatggcag
     3241 ttaaaagaat tcggtcaact gtggatgaaa aagaacaaaa acaacttctc atggatttgg
     3301 atgtagtaat gcggagtagt gattgcccat acattgttca gttctatggt gcactcttca
     3361 gagagggcga ctgttggatc tgtatggagc tcatgtctac ctcgttcgat aagttttaca
     3421 aatatgtata tagtgtgtta gatgacgtta ttccggaaga gatcttaggc aaaatcactt
     3481 tagcaactgt gaaagcacta aaccacttaa aagaaaactt gaaaattatt cacagagaca
     3541 tcaaaccttc caatattctt ctggacagaa gtggaaatat aaagctctgt gatttcggca
     3601 tcagtggaca gcttgtggac tctattgcca agacaagaga tgctgggtgt aggccgtata
     3661 tggcacctga aagaatagac ccaagtgcat caagacaagg gtatgatgtc cgctctgatg
     3721 tctggagttt ggggatcaca ttgtacgagt tggccacagg ccgatttcct tatccaaagt
     3781 ggaatagtgt atttgatcag ctaacacaag tggtgaaagg agaccctccg cagctgagta
     3841 attctgaaga aagggagttc tcccccagtt tcatcaactt tgtcaacttg tgccttacga
     3901 aggatgaatc caaaaggcca aagtataaag agcttctgaa acatcccttt attttgatgt
     3961 atgaagcacg tactgtagag gtcgcatgct atgtttgtaa aatcctggat cagatgccag
     4021 ccactcccag ctcgcccatg tatgtcgacc accatcacca tcaccattag gatcccgcga
     4081 ctctagataa ttctaccggg taggggaggc gcttttccca aggcagtctg gagcatgcgc
     4141 tttagcagcc ccgctgggca cttggcgcta cacaagtggc ctctggcctc gcacacattc
     4201 cacatccacc ggtaggcgcc aaccggctcc gttctttggt ggccccttcg cgccaccttc
     4261 tactcctccc ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg
     4321 acaaatggaa gtagcacgtc tcactagtct cgtgcagatg gacagcaccg ctgagcaatg
     4381 gaagcgggta ggcctttggg gcagcggcca atagcagctt tgctccttcg ctttctgggc
     4441 tcagaggctg ggaaggggtg ggtccggggg cgggctcagg ggcgggctca ggggcggggc
     4501 gggcgcccga aggtcctccg gaggcccggc attctgcacg cttcaaaagc gcacgtctgc
     4561 cgcgctgttc tcctcttcct catctccggg cctttcgacc tgcagcccaa gcttaccatg
     4621 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc
     4681 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc
     4741 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc
     4801 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc
     4861 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc
     4921 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc
     4981 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc
     5041 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag
     5101 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac
     5161 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgaccg
     5221 cgtctggaac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     5281 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     5341 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     5401 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     5461 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     5521 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     5581 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc
     5641 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     5701 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     5761 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     5821 tggaattaat tctgcagtcg agacctagaa aaacatggag caatcacaag tagcaataca
     5881 gcagctacca atgctgattg tgcctggcta gaagcacaag aggaggagga ggtgggtttt
     5941 ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
     6001 cactttttaa aagaaaagag gggactggaa gggctaattc actcccaacg aagacaagat
     6061 atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattagca gaactacaca
     6121 ccagggccag gggtcagata tccactgacc tttggatggt gctacaagct agtaccagtt
     6181 gagccagata aggtagaaga ggccaataaa ggagagaaca ccagcttgtt acaccctgtg
     6241 agcctgcatg ggatggatga cccggagaga gaagtgttag agtggaggtt tgacagccgc
     6301 ctagcatttc atcacgtggc ccgagagctg catccggagt acttcaagaa ctgctgatat
     6361 cgagcttgct acaagggact ttccgctggg gactttccag ggaggcgtgg cctgggcggg
     6421 actggggagt ggcgagccct cagatcctgc atataagcag ctgctttttg cctgtactgg
     6481 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     6541 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     6601 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     6661 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga
     6721 gagtgagagg ccttgacatt gctagcgttt taccgtcgac ctctagctag agcttggcgt
     6781 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     6841 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     6901 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     6961 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     7021 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     7081 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     7141 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     7201 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     7261 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     7321 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     7381 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     7441 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     7501 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     7561 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     7621 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     7681 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     7741 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     7801 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     7861 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     7921 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     7981 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     8041 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     8101 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     8161 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     8221 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     8281 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     8341 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     8401 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     8461 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     8521 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     8581 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     8641 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     8701 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     8761 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     8821 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     8881 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     8941 acgtcgacgg atcgggagat caacttgttt attgcagctt ataatggtta caaataaagc
     9001 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     9061 tccaaactca tcaatgtatc ttatcatgtc tggatcaact ggataactca agctaaccaa
     9121 aatcatccca aacttcccac cccataccct attaccactg ccaattacct gtggtttcat
     9181 ttactctaaa cctgtgattc ctctgaatta ttttcatttt aaagaaattg tatttgttaa
     9241 atatgtacta caaacttagt agtttttaaa gaaattgtat ttgttaaata tgtactacaa
     9301 acttagtagt
//
