LOCUS       V030031                 8621 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030031
VERSION     V030031
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..3377
                     /label="GREM1(NM_013372)"
                     /note="GREM1(NM_013372)"
                     /gene="GREM1"
     promoter        3409..3908
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             3929..4528
                     /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    4542..5130
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5337..5970
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6099..6115)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6123..6139
                     /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(6147..6177)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6192..6213
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6501..7089)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7260..8120)
                     /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(8121..8225)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    8273..8407
                     /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 ccaccatgag ccgcacagcc tacacggtgg gagccctgct tctcctcttg gggaccctgc
     2881 tgccggctgc tgaagggaaa aagaaagggt cccaaggtgc catccccccg ccagacaagg
     2941 cccagcacaa tgactcagag cagactcagt cgccccagca gcctggctcc aggaaccggg
     3001 ggcggggcca agggcggggc actgccatgc ccggggagga ggtgctggag tccagccaag
     3061 aggccctgca tgtgacggag cgcaaatacc tgaagcgaga ctggtgcaaa acccagccgc
     3121 ttaagcagac catccacgag gaaggctgca acagtcgcac catcatcaac cgcttctgtt
     3181 acggccagtg caactctttc tacatcccca ggcacatccg gaaggaggaa ggttcctttc
     3241 agtcctgctc cttctgcaag cccaagaaat tcactaccat gatggtcaca ctcaactgcc
     3301 ctgaactaca gccacctacc aagaagaaga gagtcacacg tgtgaagcag tgtcgttgca
     3361 tatccatcga tttggattaa ggatcccgcg actctagata attctaccgg gtaggggagg
     3421 cgcttttccc aaggcagtct ggagcatgcg ctttagcagc cccgctgggc acttggcgct
     3481 acacaagtgg cctctggcct cgcacacatt ccacatccac cggtaggcgc caaccggctc
     3541 cgttctttgg tggccccttc gcgccacctt ctactcctcc cctagtcagg aagttccccc
     3601 ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga agtagcacgt ctcactagtc
     3661 tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt aggcctttgg ggcagcggcc
     3721 aatagcagct ttgctccttc gctttctggg ctcagaggct gggaaggggt gggtccgggg
     3781 gcgggctcag gggcgggctc aggggcgggg cgggcgcccg aaggtcctcc ggaggcccgg
     3841 cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt ctcctcttcc tcatctccgg
     3901 gcctttcgac ctgcagccca agcttaccat gaccgagtac aagcccacgg tgcgcctcgc
     3961 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     4021 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga
     4081 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     4141 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     4201 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     4261 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     4321 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     4381 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     4441 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     4501 gtgcatgacc cgcaagcccg gtgcctgacc gcgtctggaa caatcaacct ctggattaca
     4561 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     4621 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     4681 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     4741 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     4801 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     4861 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     4921 tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga
     4981 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     5041 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     5101 gtcggatctc cctttgggcc gcctccccgc ctggaattaa ttctgcagtc gagacctaga
     5161 aaaacatgga gcaatcacaa gtagcaatac agcagctacc aatgctgatt gtgcctggct
     5221 agaagcacaa gaggaggagg aggtgggttt tccagtcaca cctcaggtac ctttaagacc
     5281 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaaga ggggactgga
     5341 agggctaatt cactcccaac gaagacaaga tatccttgat ctgtggatct accacacaca
     5401 aggctacttc cctgattagc agaactacac accagggcca ggggtcagat atccactgac
     5461 ctttggatgg tgctacaagc tagtaccagt tgagccagat aaggtagaag aggccaataa
     5521 aggagagaac accagcttgt tacaccctgt gagcctgcat gggatggatg acccggagag
     5581 agaagtgtta gagtggaggt ttgacagccg cctagcattt catcacgtgg cccgagagct
     5641 gcatccggag tacttcaaga actgctgata tcgagcttgc tacaagggac tttccgctgg
     5701 ggactttcca gggaggcgtg gcctgggcgg gactggggag tggcgagccc tcagatcctg
     5761 catataagca gctgcttttt gcctgtactg ggtctctctg gttagaccag atctgagcct
     5821 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     5881 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     5941 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt
     6001 atttataact tgcaaagaaa tgaatatcag agagtgagag gccttgacat tgctagcgtt
     6061 ttaccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     6121 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     6181 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     6241 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     6301 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     6361 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     6421 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     6481 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     6541 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6601 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6661 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     6721 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6781 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6841 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6901 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     6961 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     7021 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     7081 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     7141 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     7201 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     7261 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     7321 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     7381 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     7441 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     7501 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     7561 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     7621 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     7681 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     7741 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     7801 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7861 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     7921 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     7981 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     8041 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     8101 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     8161 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     8221 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tcaacttgtt
     8281 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     8341 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     8401 ctggatcaac tggataactc aagctaacca aaatcatccc aaacttccca ccccataccc
     8461 tattaccact gccaattacc tgtggtttca tttactctaa acctgtgatt cctctgaatt
     8521 attttcattt taaagaaatt gtatttgtta aatatgtact acaaacttag tagtttttaa
     8581 agaaattgta tttgttaaat atgtactaca aacttagtag t
//
