LOCUS       V022355                 8735 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022355
VERSION     V022355
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..3936
                     /label="WNT1(NM_005430)"
                     /note="WNT1(NM_005430)"
                     /gene="WNT1"
     CDS             3958..3981
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             3991..4044
                     /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             4075..4782
                     /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    4789..5377
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5451..6084
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6213..6229)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6237..6253
                     /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(6261..6291)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6306..6327
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6615..7203)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7374..8234)
                     /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(8235..8339)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    8387..8521
                     /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 gccaccatgg ggctctgggc gctgttgcct ggctgggttt ctgctacgct gctgctggcg
     2881 ctggccgctc tgcccgcagc cctggctgcc aacagcagtg gccgatggtg gggtattgtg
     2941 aacgtagcct cctccacgaa cctgcttaca gactccaaga gtctgcaact ggtactcgag
     3001 cccagtctgc agctgttgag ccgcaaacag cggcgtctga tacgccaaaa tccggggatc
     3061 ctgcacagcg tgagtggggg gctgcagagt gccgtgcgcg agtgcaagtg gcagttccgg
     3121 aatcgccgct ggaactgtcc cactgctcca gggccccacc tcttcggcaa gatcgtcaac
     3181 cgaggctgtc gagaaacggc gtttatcttc gctatcacct ccgccggggt cacccattcg
     3241 gtggcgcgct cctgctcaga aggttccatc gaatcctgca cgtgtgacta ccggcggcgc
     3301 ggccccgggg gccccgactg gcactggggg ggctgcagcg acaacattga cttcggccgc
     3361 ctcttcggcc gggagttcgt ggactccggg gagaaggggc gggacctgcg cttcctcatg
     3421 aaccttcaca acaacgaggc aggccgtacg accgtattct ccgagatgcg ccaggagtgc
     3481 aagtgccacg ggatgtccgg ctcatgcacg gtgcgcacgt gctggatgcg gctgcccacg
     3541 ctgcgcgccg tgggcgatgt gctgcgcgac cgcttcgacg gcgcctcgcg cgtcctgtac
     3601 ggcaaccgcg gcagcaaccg cgcttcgcgg gcggagctgc tgcgcctgga gccggaagac
     3661 ccggcccaca aaccgccctc cccccacgac ctcgtctact tcgagaaatc gcccaacttc
     3721 tgcacgtaca gcggacgcct gggcacagca ggcacggcag ggcgcgcctg taacagctcg
     3781 tcgcccgcgc tggacggctg cgagctgctc tgctgcggca ggggccaccg cacgcgcacg
     3841 cagcgcgtca ccgagcgctg caactgcacc ttccactggt gctgccacgt cagctgccgc
     3901 aactgcacgc acacgcgcgt actgcacgag tgtctggaat tcggaggagg gggaagcgac
     3961 tacaaggacg acgatgacaa gggcagcggt gagggcagag gaagtcttct aacatgcggt
     4021 gacgtggagg agaatcccgg cccttccgga atggagagcg acgagagcgg cctggtgagc
     4081 aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg
     4141 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
     4201 gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg
     4261 gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac
     4321 atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac
     4381 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc
     4441 atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag
     4501 aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag
     4561 ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag
     4621 accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag
     4681 ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag
     4741 ggccgccact ccaccggcgg catggacgag ctgtacaagt aggtcgacaa tcaacctctg
     4801 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     4861 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     4921 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     4981 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     5041 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     5101 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     5161 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc
     5221 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     5281 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     5341 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa gaccaatgac
     5401 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aagaggggac tggaagggct
     5461 aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta
     5521 cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac tgacctttgg
     5581 atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca ataaaggaga
     5641 gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt
     5701 gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag agctgcatcc
     5761 ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg ctggggactt
     5821 tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat cctgcatata
     5881 agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga gcctgggagc
     5941 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     6001 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     6061 agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat
     6121 aacttgcaaa gaaatgaata tcagagagtg agaggccttg acattgctag cgttttaccg
     6181 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     6241 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     6301 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     6361 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     6421 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     6481 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     6541 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     6601 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     6661 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6721 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6781 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     6841 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6901 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6961 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     7021 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     7081 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     7141 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     7201 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     7261 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     7321 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     7381 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     7441 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     7501 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     7561 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     7621 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     7681 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     7741 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7801 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7861 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7921 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7981 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     8041 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     8101 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     8161 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     8221 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     8281 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     8341 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatcaact tgtttattgc
     8401 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     8461 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat
     8521 caactggata actcaagcta accaaaatca tcccaaactt cccaccccat accctattac
     8581 cactgccaat tacctgtggt ttcatttact ctaaacctgt gattcctctg aattattttc
     8641 attttaaaga aattgtattt gttaaatatg tactacaaac ttagtagttt ttaaagaaat
     8701 tgtatttgtt aaatatgtac tacaaactta gtagt
//
