LOCUS       V022357                 8687 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022357
VERSION     V022357
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..3888
                     /label="WNT11(NM_004626)"
                     /note="WNT11(NM_004626)"
                     /gene="WNT11"
     CDS             3910..3933
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             3943..3996
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from&nbsp;Thosea asigna virus capsid
                     protein"
                     /protein_id=""
                     /transl_table=1
                     /translation="EGRGSLLTCGDVEENPGP"
                     /label="T2A"
     CDS             4027..4734
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK*"
                     /label="mCherry"
     misc_feature    4741..5329
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5403..6036
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6165..6181)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6189..6205
                     /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(6213..6243)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6258..6279
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6567..7155)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7326..8186)
                     /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(8187..8291)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    8339..8473
                     /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 gggcgcggcc gcaggtctgc gaggcgctgc tcttcgccct ggcgctccag
     2881 accggcgtgt gctatggcat caagtggctg gcgctgtcca agacaccatc ggccctggca
     2941 ctgaaccaga cgcaacactg caagcagctg gagggtctgg tgtctgcaca ggtgcagctg
     3001 tgccgcagca acctggagct catgcacacg gtggtgcacg ccgcccgcga ggtcatgaag
     3061 gcctgtcgcc gggcctttgc cgacatgcgc tggaactgct cctccattga gctcgccccc
     3121 aactatttgc ttgacctgga gagagggacc cgggagtcgg ccttcgtgta tgcgctgtcg
     3181 gccgccgcca tcagccacgc catcgcccgg gcctgcacct ccggcgacct gcccggctgc
     3241 tcctgcggcc ccgtcccagg tgagccaccc gggcccggga accgctgggg aggatgtgcg
     3301 gacaacctca gctacgggct cctcatgggg gccaagtttt ccgatgctcc tatgaaggtg
     3361 aaaaaaacag gatcccaagc caataaactg atgcgtctac acaacagtga agtggggaga
     3421 caggctctgc gcgcctctct ggaaatgaag tgtaagtgcc atggggtgtc tggctcctgc
     3481 tccatccgca cctgctggaa ggggctgcag gagctgcagg atgtggctgc tgacctcaag
     3541 acccgatacc tgtcggccac caaggtagtg caccgaccca tgggcacccg caagcacctg
     3601 gtgcccaagg acctggatat ccggcctgtg aaggactcgg aactcgtcta tctgcagagc
     3661 tcacctgact tctgcatgaa gaatgagaag gtgggctccc acgggacaca agacaggcag
     3721 tgcaacaaga catccaacgg aagcgacagc tgcgacctta tgtgctgcgg gcgtggctac
     3781 aacccctaca cagaccgcgt ggtcgagcgg tgccactgta agtaccactg gtgctgctac
     3841 gtcacctgcc gcaggtgtga gcgtaccgtg gagcgctatg tctgcaagga attcggagga
     3901 gggggaagcg actacaagga cgacgatgac aagggcagcg gtgagggcag aggaagtctt
     3961 ctaacatgcg gtgacgtgga ggagaatccc ggcccttccg gaatggagag cgacgagagc
     4021 ggcctggtga gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc
     4081 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
     4141 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
     4201 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
     4261 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
     4321 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
     4381 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     4441 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
     4501 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
     4561 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
     4621 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     4681 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaggtcgac
     4741 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     4801 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     4861 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     4921 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     4981 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     5041 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     5101 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     5161 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     5221 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     5281 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggtaccttt
     5341 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagagggg
     5401 actggaaggg ctaattcact cccaacgaag acaagatatc cttgatctgt ggatctacca
     5461 cacacaaggc tacttccctg attagcagaa ctacacacca gggccagggg tcagatatcc
     5521 actgaccttt ggatggtgct acaagctagt accagttgag ccagataagg tagaagaggc
     5581 caataaagga gagaacacca gcttgttaca ccctgtgagc ctgcatggga tggatgaccc
     5641 ggagagagaa gtgttagagt ggaggtttga cagccgccta gcatttcatc acgtggcccg
     5701 agagctgcat ccggagtact tcaagaactg ctgatatcga gcttgctaca agggactttc
     5761 cgctggggac tttccaggga ggcgtggcct gggcgggact ggggagtggc gagccctcag
     5821 atcctgcata taagcagctg ctttttgcct gtactgggtc tctctggtta gaccagatct
     5881 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     5941 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     6001 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta
     6061 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggcct tgacattgct
     6121 agcgttttac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     6181 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     6241 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     6301 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     6361 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     6421 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     6481 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     6541 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     6601 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     6661 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     6721 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     6781 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6841 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6901 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6961 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     7021 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     7081 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     7141 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     7201 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     7261 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     7321 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     7381 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     7441 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     7501 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     7561 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     7621 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     7681 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     7741 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     7801 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7861 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7921 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7981 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     8041 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     8101 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     8161 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     8221 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     8281 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatcaa
     8341 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     8401 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     8461 tcatgtctgg atcaactgga taactcaagc taaccaaaat catcccaaac ttcccacccc
     8521 ataccctatt accactgcca attacctgtg gtttcattta ctctaaacct gtgattcctc
     8581 tgaattattt tcattttaaa gaaattgtat ttgttaaata tgtactacaa acttagtagt
     8641 ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagt
//
