LOCUS       V023331                 8908 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023331
VERSION     V023331
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..3641
                     /label="RSPO3(NM_032784)"
                     /note="RSPO3(NM_032784)"
                     /gene="RSPO3"
     CDS             3648..3665
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     promoter        3696..4195
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4216..4815
                     /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    4829..5417
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5624..6257
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6386..6402)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6410..6426
                     /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(6434..6464)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6479..6500
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6788..7376)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7547..8407)
                     /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(8408..8512)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    8560..8694
                     /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 ccaccatgca cttgcgactg atttcttggc tttttatcat tttgaacttt atggaataca
     2881 tcggcagcca aaacgcctcc cggggaaggc gccagcgaag aatgcatcct aacgttagtc
     2941 aaggctgcca aggaggctgt gcaacatgct cagattacaa tggatgtttg tcatgtaagc
     3001 ccagactatt ttttgctctg gaaagaattg gcatgaagca gattggagta tgtctctctt
     3061 catgtccaag tggatattat ggaactcgat atccagatat aaataagtgt acaaaatgca
     3121 aagctgactg tgatacctgt ttcaacaaaa atttctgcac aaaatgtaaa agtggatttt
     3181 acttacacct tggaaagtgc cttgacaatt gcccagaagg gttggaagcc aacaaccata
     3241 ctatggagtg tgtcagtatt gtgcactgtg aggtcagtga atggaatcct tggagtccat
     3301 gcacgaagaa gggaaaaaca tgtggcttca aaagagggac tgaaacacgg gtccgagaaa
     3361 taatacagca tccttcagca aagggtaacc tgtgtccccc aacaaatgag acaagaaagt
     3421 gtacagtgca aaggaagaag tgtcagaagg gagaacgagg aaaaaaagga agggagagga
     3481 aaagaaaaaa acctaataaa ggagaaagta aagaagcaat acctgacagc aaaagtctgg
     3541 aatccagcaa agaaatccca gagcaacgag aaaacaaaca gcagcagaag aagcgaaaag
     3601 tccaagataa acagaaatcg gtatcagtca gcactgtaca cgaattccat catcaccatc
     3661 accattagga tcccgcgact ctagataatt ctaccgggta ggggaggcgc ttttcccaag
     3721 gcagtctgga gcatgcgctt tagcagcccc gctgggcact tggcgctaca caagtggcct
     3781 ctggcctcgc acacattcca catccaccgg taggcgccaa ccggctccgt tctttggtgg
     3841 ccccttcgcg ccaccttcta ctcctcccct agtcaggaag ttcccccccg ccccgcagct
     3901 cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc actagtctcg tgcagatgga
     3961 cagcaccgct gagcaatgga agcgggtagg cctttggggc agcggccaat agcagctttg
     4021 ctccttcgct ttctgggctc agaggctggg aaggggtggg tccgggggcg ggctcagggg
     4081 cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat tctgcacgct
     4141 tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca tctccgggcc tttcgacctg
     4201 cagcccaagc ttaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     4261 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     4321 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     4381 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     4441 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     4501 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg
     4561 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
     4621 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg
     4681 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
     4741 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
     4801 aagcccggtg cctgaccgcg tctggaacaa tcaacctctg gattacaaaa tttgtgaaag
     4861 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     4921 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     4981 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     5041 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     5101 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     5161 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     5221 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     5281 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     5341 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     5401 ttgggccgcc tccccgcctg gaattaattc tgcagtcgag acctagaaaa acatggagca
     5461 atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga agcacaagag
     5521 gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat gacttacaag
     5581 gcagctgtag atcttagcca ctttttaaaa gaaaagaggg gactggaagg gctaattcac
     5641 tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct
     5701 gattagcaga actacacacc agggccaggg gtcagatatc cactgacctt tggatggtgc
     5761 tacaagctag taccagttga gccagataag gtagaagagg ccaataaagg agagaacacc
     5821 agcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga agtgttagag
     5881 tggaggtttg acagccgcct agcatttcat cacgtggccc gagagctgca tccggagtac
     5941 ttcaagaact gctgatatcg agcttgctac aagggacttt ccgctgggga ctttccaggg
     6001 aggcgtggcc tgggcgggac tggggagtgg cgagccctca gatcctgcat ataagcagct
     6061 gctttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     6121 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     6181 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     6241 gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc
     6301 aaagaaatga atatcagaga gtgagaggcc ttgacattgc tagcgtttta ccgtcgacct
     6361 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     6421 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     6481 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     6541 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     6601 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6661 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6721 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6781 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6841 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6901 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6961 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     7021 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     7081 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     7141 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     7201 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     7261 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     7321 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     7381 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     7441 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     7501 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     7561 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     7621 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     7681 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     7741 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     7801 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     7861 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7921 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7981 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     8041 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     8101 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     8161 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     8221 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     8281 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     8341 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     8401 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     8461 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     8521 cccgaaaagt gccacctgac gtcgacggat cgggagatca acttgtttat tgcagcttat
     8581 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     8641 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcaactgg
     8701 ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc cataccctat taccactgcc
     8761 aattacctgt ggtttcattt actctaaacc tgtgattcct ctgaattatt ttcattttaa
     8821 agaaattgta tttgttaaat atgtactaca aacttagtag tttttaaaga aattgtattt
     8881 gttaaatatg tactacaaac ttagtagt
//
