LOCUS       V029891                 8993 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029891
VERSION     V029891
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             2815..3603
                     /label="KCTD2(NM_015353)"
                     /note="KCTD2(NM_015353)"
                     /gene="KCTD2"
     CDS             3604..3633
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     misc_feature    3664..4236
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES"
     CDS             4238..4948
                     /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"
     misc_feature    4962..5550
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5757..6390
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6518..6534)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6542..6558
                     /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(6566..6596)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6611..6632
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6920..7508)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7679..8539)
                     /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(8540..8644)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    8692..8826
                     /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 tctatttccg gtgaattcgc caccatggcg
     2821 gaactgcagc tggacccggc gatggcgggg ctgggagggg gcggcgggag tggggtgggc
     2881 gacgggggtg gcccagtccg cgggcccccc agcccacgcc cggctggccc cacgccccgc
     2941 gggcacggcc gcccggctgc cgccgtcgcg cagccgctgg agccgggtcc cggaccaccc
     3001 gagcgggcag ggggcggcgg cgcggcccgc tgggtcaggc tgaacgtggg aggcacctac
     3061 ttcgtgacca ccagacagac cttaggccgg gagcccaagt catttctctg ccgcctctgc
     3121 tgccaggagg acccggagct ggactcagac aaggatgaga caggagccta tctgattgac
     3181 agggacccca cctactttgg tcctatcctc aactacctcc gccacgggaa actcatcatc
     3241 actaaggagt tggcagaaga aggtgtgctg gaggaagcgg agttttacaa catcgcgtcc
     3301 cttgtgcggc tggttaagga aaggatacgg gacaatgaga acagaacttc acaaggcccc
     3361 gtgaagcacg tgtacagagt cctgcagtgt caggaagaag agctcacgca gatggtgtcc
     3421 acgatgtccg acggctggaa attcgaacag ctcatcagca tcggatcttc ctataactac
     3481 ggcaatgagg atcaggcaga attcctctgt gttgtctcca gagaactaaa taattctacc
     3541 aatggcatcg tcatagagcc gagcgaaaag gcgaagattc ttcaggagag aggatcgcgg
     3601 atggagcaga agctgatctc agaggaggac ctgtgactag ttctagagcg gccgcggatc
     3661 ccgcccctct ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg
     3721 gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc
     3781 ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa
     3841 aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag
     3901 acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg
     3961 cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg
     4021 ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctcacctcaa gcgtattcaa
     4081 caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg
     4141 gtgcacatgc tttacatgtg tttagtcgag gttaaaaaac gtctaggccc cccgaaccac
     4201 ggggacgtgg ttttcctttg aaaaacacga tgataatatg gtgagcaagg gcgaggagga
     4261 taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa
     4321 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
     4381 cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc
     4441 tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt
     4501 gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
     4561 cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa
     4621 gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg
     4681 ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca
     4741 gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc
     4801 caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc
     4861 ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac
     4921 cggcggcatg gacgagctgt acaagtgaac gcgtctggaa caatcaacct ctggattaca
     4981 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     5041 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     5101 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     5161 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     5221 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     5281 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     5341 tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga
     5401 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     5461 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     5521 gtcggatctc cctttgggcc gcctccccgc ctggaattaa ttctgcagtc gagacctaga
     5581 aaaacatgga gcaatcacaa gtagcaatac agcagctacc aatgctgatt gtgcctggct
     5641 agaagcacaa gaggaggagg aggtgggttt tccagtcaca cctcaggtac ctttaagacc
     5701 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaaga ggggactgga
     5761 agggctaatt cactcccaac gaagacaaga tatccttgat ctgtggatct accacacaca
     5821 aggctacttc cctgattagc agaactacac accagggcca ggggtcagat atccactgac
     5881 ctttggatgg tgctacaagc tagtaccagt tgagccagat aaggtagaag aggccaataa
     5941 aggagagaac accagcttgt tacaccctgt gagcctgcat gggatggatg acccggagag
     6001 agaagtgtta gagtggaggt ttgacagccg cctagcattt catcacgtgg cccgagagct
     6061 gcatccggag tacttcaaga actgctgata tcgagcttgc tacaagggac tttccgctgg
     6121 ggactttcca gggaggcgtg gcctgggcgg gactggggag tggcgagccc tcagatcctg
     6181 catataagca gctgcttttt gcctgtactg ggtctctctg gttagaccag atctgagcct
     6241 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     6301 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     6361 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt
     6421 atttataact tgcaaagaaa tgaatatcag agagtgagag gccttgacat tgctagcgtt
     6481 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     6541 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     6601 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     6661 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     6721 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     6781 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     6841 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6901 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6961 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     7021 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     7081 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     7141 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     7201 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     7261 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     7321 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     7381 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     7441 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     7501 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     7561 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     7621 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     7681 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     7741 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     7801 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     7861 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7921 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7981 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     8041 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     8101 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     8161 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     8221 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     8281 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     8341 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     8401 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     8461 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     8521 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     8581 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     8641 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat caacttgttt
     8701 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     8761 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     8821 tggatcaact ggataactca agctaaccaa aatcatccca aacttcccac cccataccct
     8881 attaccactg ccaattacct gtggtttcat ttactctaaa cctgtgattc ctctgaatta
     8941 ttttcatttt aaagaaattg tatttgttaa atatgtacta caaacttagt agt
//
