LOCUS       V019471                 9813 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019471
VERSION     V019471
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..4503
                     /label="YTHDF1(NM_017798)"
                     /note="YTHDF1(NM_017798)"
                     /gene="YTHDF1"
     CDS             4504..4569
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4601..5100
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5121..5720
                     /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    5734..6322
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6529..7162
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7291..7307)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7315..7331
                     /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(7339..7369)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7384..7405
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7693..8281)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8452..9312)
                     /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(9313..9417)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9465..9599
                     /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 gccaccatgt cggccaccag cgtggacacc cagagaacaa aaggacaaga taataaagta
     2881 caaaatggtt cgttacatca gaaggataca gttcatgaca atgactttga gccctacctt
     2941 actggacagt caaatcagag taacagttac ccctcaatga gcgaccccta cctgtccagc
     3001 tattacccgc cgtccattgg atttccttac tccctcaatg aggctccgtg gtctactgca
     3061 ggggaccctc cgattccata cctcaccacc tacggacagc tcagtaacgg agaccatcat
     3121 tttatgcacg atgctgtttt tgggcagcct gggggcctgg ggaacaacat ctatcagcac
     3181 aggttcaatt ttttccctga aaaccctgcg ttctcagcat gggggacaag tgggtctcaa
     3241 ggtcagcaga cccagagctc cgcgtatggg agcagctaca cctacccccc gagctccctg
     3301 ggtggcacgg tggttgatgg gcagccaggc tttcacagcg acaccctcag caaggccccc
     3361 gggatgaaca gcctggagca gggcatggtt ggcctgaaga ttggggacgt cagctcctcc
     3421 gccgtcaaga cggtgggctc tgtcgtcagc agcgtggcac tgactggtgt cctttctggc
     3481 aacggtggga caaatgtgaa catgccagtt tcaaagccga cctcgtgggc tgccattgcc
     3541 agcaagcctg caaaaccaca gcctaaaatg aaaacaaaga gcgggcctgt catggggggt
     3601 gggctgcccc ctccacccat aaagcataac atggacattg gcacctggga taacaagggg
     3661 cctgtgccga aggccccagt cccccagcag gcaccctctc cacaggctgc cccacagccc
     3721 cagcaggtgg ctcagcctct cccagcacag cccccagctt tggctcaacc gcagtatcag
     3781 agccctcagc agccacccca gacccgctgg gttgccccac gcaacagaaa cgcggcgttt
     3841 gggcagagcg gaggggctgg cagcgatagc aactctcctg gaaacgtcca gcctaattct
     3901 gcccccagcg tcgaatccca ccccgtcctt gaaaaactga aggctgctca cagctacaac
     3961 ccgaaagagt ttgagtggaa tctgaaaagc gggcgtgtgt tcatcatcaa gagctactct
     4021 gaggacgaca tccaccgctc cattaagtac tccatctggt gtagcacaga gcacggcaac
     4081 aagcgcctgg acagcgcctt ccgctgcatg agcagcaagg ggcccgtcta cctgctcttc
     4141 agcgtcaatg ggagtgggca tttttgtggg gtggccgaga tgaagtcccc cgtggactac
     4201 ggcaccagtg ccggggtctg gtctcaggac aagtggaagg ggaagtttga tgtccagtgg
     4261 atttttgtta aggatgtacc caataaccag ctccggcaca tcaggctgga gaataacgac
     4321 aacaaaccgg tcacaaactc ccgggacacc caggaggtgc ccttagaaaa agccaagcaa
     4381 gtgctgaaaa ttatcagttc ctacaagcac acaacctcca tcttcgacga ctttgctcac
     4441 tacgagaagc gccaggagga ggaggaggtg gtgcgcaagg aacggcagag tcgaaacaaa
     4501 caagactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac
     4561 gatgacaagt agggatcccg cgactctaga taattctacc gggtagggga ggcgcttttc
     4621 ccaaggcagt ctggagcatg cgctttagca gccccgctgg gcacttggcg ctacacaagt
     4681 ggcctctggc ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt
     4741 ggtggcccct tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg
     4801 cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag
     4861 atggacagca ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag
     4921 ctttgctcct tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc
     4981 aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc
     5041 acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc gggcctttcg
     5101 acctgcagcc caagcttacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg
     5161 acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
     5221 gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
     5281 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
     5341 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
     5401 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg
     5461 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc
     5521 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg
     5581 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
     5641 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
     5701 cccgcaagcc cggtgcctga ccgcgtctgg aacaatcaac ctctggatta caaaatttgt
     5761 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     5821 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     5881 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     5941 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     6001 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     6061 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     6121 tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc
     6181 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     6241 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     6301 tccctttggg ccgcctcccc gcctggaatt aattctgcag tcgagaccta gaaaaacatg
     6361 gagcaatcac aagtagcaat acagcagcta ccaatgctga ttgtgcctgg ctagaagcac
     6421 aagaggagga ggaggtgggt tttccagtca cacctcaggt acctttaaga ccaatgactt
     6481 acaaggcagc tgtagatctt agccactttt taaaagaaaa gaggggactg gaagggctaa
     6541 ttcactccca acgaagacaa gatatccttg atctgtggat ctaccacaca caaggctact
     6601 tccctgatta gcagaactac acaccagggc caggggtcag atatccactg acctttggat
     6661 ggtgctacaa gctagtacca gttgagccag ataaggtaga agaggccaat aaaggagaga
     6721 acaccagctt gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtgt
     6781 tagagtggag gtttgacagc cgcctagcat ttcatcacgt ggcccgagag ctgcatccgg
     6841 agtacttcaa gaactgctga tatcgagctt gctacaaggg actttccgct ggggactttc
     6901 cagggaggcg tggcctgggc gggactgggg agtggcgagc cctcagatcc tgcatataag
     6961 cagctgcttt ttgcctgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     7021 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     7081 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     7141 tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa
     7201 cttgcaaaga aatgaatatc agagagtgag aggccttgac attgctagcg ttttaccgtc
     7261 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     7321 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     7381 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     7441 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     7501 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     7561 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     7621 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     7681 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     7741 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     7801 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     7861 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     7921 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     7981 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     8041 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     8101 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     8161 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     8221 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     8281 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     8341 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     8401 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     8461 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     8521 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     8581 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     8641 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     8701 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     8761 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     8821 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     8881 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     8941 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     9001 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     9061 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     9121 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     9181 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     9241 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     9301 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     9361 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     9421 atttccccga aaagtgccac ctgacgtcga cggatcggga gatcaacttg tttattgcag
     9481 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     9541 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatca
     9601 actggataac tcaagctaac caaaatcatc ccaaacttcc caccccatac cctattacca
     9661 ctgccaatta cctgtggttt catttactct aaacctgtga ttcctctgaa ttattttcat
     9721 tttaaagaaa ttgtatttgt taaatatgta ctacaaactt agtagttttt aaagaaattg
     9781 tatttgttaa atatgtacta caaacttagt agt
//
