LOCUS       V022911                 8881 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022911
VERSION     V022911
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..3614
                     /label="RSPO1(NM_001038633)"
                     /note="RSPO1(NM_001038633)"
                     /gene="RSPO1"
     CDS             3621..3638
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     promoter        3669..4168
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4189..4788
                     /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    4802..5390
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5597..6230
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6359..6375)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6383..6399
                     /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(6407..6437)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6452..6473
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6761..7349)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7520..8380)
                     /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(8381..8485)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    8533..8667
                     /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 ccaccatgcg gcttgggctg tgtgtggtgg ccctggttct gagctggacg cacctcacca
     2881 tcagcagccg ggggatcaag gggaaaaggc agaggcggat cagtgccgag gggagccagg
     2941 cctgtgccaa aggctgtgag ctctgctctg aagtcaacgg ctgcctcaag tgctcaccca
     3001 agctgttcat cctgctggag aggaacgaca tccgccaggt gggcgtctgc ttgccgtcct
     3061 gcccacctgg atacttcgac gcccgcaacc ccgacatgaa caagtgcatc aaatgcaaga
     3121 tcgagcactg tgaggcctgc ttcagccata acttctgcac caagtgtaag gagggcttgt
     3181 acctgcacaa gggccgctgc tatccagctt gtcccgaggg ctcctcagct gccaatggca
     3241 ccatggagtg cagtagtcct gcgcaatgtg aaatgagcga gtggtctccg tgggggccct
     3301 gctccaagaa gcagcagctc tgtggtttcc ggaggggctc cgaggagcgg acacgcaggg
     3361 tgctacatgc ccctgtgggg gaccatgctg cctgctctga caccaaggag acccggaggt
     3421 gcacagtgag gagagtgccg tgtcctgagg ggcagaagag gaggaaggga ggccagggcc
     3481 ggcgggagaa tgccaacagg aacctggcca ggaaggagag caaggaggcg ggtgctggct
     3541 ctcgaagacg caaggggcag caacagcagc agcagcaagg gacagtgggg ccactcacat
     3601 ctgcagggcc tgccgaattc catcatcacc atcaccatta ggatcccgcg actctagata
     3661 attctaccgg gtaggggagg cgcttttccc aaggcagtct ggagcatgcg ctttagcagc
     3721 cccgctgggc acttggcgct acacaagtgg cctctggcct cgcacacatt ccacatccac
     3781 cggtaggcgc caaccggctc cgttctttgg tggccccttc gcgccacctt ctactcctcc
     3841 cctagtcagg aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga
     3901 agtagcacgt ctcactagtc tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt
     3961 aggcctttgg ggcagcggcc aatagcagct ttgctccttc gctttctggg ctcagaggct
     4021 gggaaggggt gggtccgggg gcgggctcag gggcgggctc aggggcgggg cgggcgcccg
     4081 aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt
     4141 ctcctcttcc tcatctccgg gcctttcgac ctgcagccca agcttaccat gaccgagtac
     4201 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
     4261 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
     4321 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
     4381 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
     4441 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
     4501 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
     4561 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
     4621 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     4681 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
     4741 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacc gcgtctggaa
     4801 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     4861 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     4921 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     4981 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     5041 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     5101 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     5161 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     5221 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     5281 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     5341 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaattaa
     5401 ttctgcagtc gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc
     5461 aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca
     5521 cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     5581 aaagaaaaga ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat
     5641 ctgtggatct accacacaca aggctacttc cctgattagc agaactacac accagggcca
     5701 ggggtcagat atccactgac ctttggatgg tgctacaagc tagtaccagt tgagccagat
     5761 aaggtagaag aggccaataa aggagagaac accagcttgt tacaccctgt gagcctgcat
     5821 gggatggatg acccggagag agaagtgtta gagtggaggt ttgacagccg cctagcattt
     5881 catcacgtgg cccgagagct gcatccggag tacttcaaga actgctgata tcgagcttgc
     5941 tacaagggac tttccgctgg ggactttcca gggaggcgtg gcctgggcgg gactggggag
     6001 tggcgagccc tcagatcctg catataagca gctgcttttt gcctgtactg ggtctctctg
     6061 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     6121 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     6181 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc
     6241 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag
     6301 gccttgacat tgctagcgtt ttaccgtcga cctctagcta gagcttggcg taatcatggt
     6361 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     6421 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     6481 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     6541 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     6601 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     6661 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6721 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6781 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6841 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6901 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6961 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     7021 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     7081 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     7141 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     7201 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     7261 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     7321 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     7381 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     7441 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     7501 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     7561 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     7621 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     7681 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     7741 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     7801 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     7861 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7921 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7981 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     8041 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     8101 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     8161 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     8221 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     8281 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     8341 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     8401 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     8461 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     8521 gatcgggaga tcaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     8581 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     8641 atcaatgtat cttatcatgt ctggatcaac tggataactc aagctaacca aaatcatccc
     8701 aaacttccca ccccataccc tattaccact gccaattacc tgtggtttca tttactctaa
     8761 acctgtgatt cctctgaatt attttcattt taaagaaatt gtatttgtta aatatgtact
     8821 acaaacttag tagtttttaa agaaattgta tttgttaaat atgtactaca aacttagtag
     8881 t
//
