LOCUS       V031390                10025 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031390
VERSION     V031390
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..4551
                     /label="YTHDF2(NM_001173128)"
                     /note="YTHDF2(NM_001173128)"
                     /gene="YTHDF2"
     CDS             4561..4587
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    4599..5185
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES2"
     CDS             5186..5980
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     misc_feature    5994..6582
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6789..7422
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7550..7566)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7574..7590
                     /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(7598..7628)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7643..7664
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7952..8540)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8711..9571)
                     /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(9572..9676)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9724..9858
                     /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 caccatgtcg
     2821 gccagcagcc tcttggagca gagaccaaaa ggtcaaggaa acaaagtaca aaatggatct
     2881 gtacatcaaa aggatggatt aaacgatgat gattttgaac cttacttgag tccacaggca
     2941 aggcccaata atgcatatac tgccatgtca gattcctact tacccagtta ctacagtccc
     3001 tccattggct tctcctattc tttgggtgaa gctgcttggt ctacgggggg tgacacagcc
     3061 atgccctact taacttctta tggacagctg agcaacggag agccccactt cctaccagat
     3121 gcaatgtttg ggcaaccagg agccctaggt agcactccat ttcttggtca gcatggtttt
     3181 aatttctttc ccagtgggat tgacttctca gcatggggaa ataacagttc tcagggacag
     3241 tctactcaga gctctggata tagtagcaat tatgcttatg cacctagctc cttaggtgga
     3301 gccatgattg atggacagtc agcttttgcc aatgagaccc tcaataaggc tcctggcatg
     3361 aatactatag accaagggat ggcagcactg aagttgggta gcacagaagt tgcaagcaat
     3421 gttccaaaag ttgtaggttc tgctgttggt agcgggtcca ttactagtaa catcgtggct
     3481 tccaatagtt tgcctccagc caccattgct cctccaaaac cagcatcttg ggctgatatt
     3541 gctagcaagc ctgcaaaaca gcaacctaaa ctgaagacca agaatggcat tgcagggtca
     3601 agtcttccgc cacccccgat aaagcataac atggatattg gaacttggga taacaagggt
     3661 cccgttgcaa aagccccctc acaggctttg gttcagaata taggtcagcc aacccagggg
     3721 tctcctcagc ctgtaggtca gcaggctaac aatagcccac cagtggctca ggcatcagta
     3781 gggcaacaga cacagccatt gcctccacct ccaccacagc ctgcccagct ttcagtccag
     3841 caacaggcag ctcagccaac ccgctgggta gcacctcgga accgtggcag tgggttcggt
     3901 cataatgggg tggatggtaa tggagtagga cagtctcagg ctggttctgg atctactcct
     3961 tcagaacccc acccagtgtt ggagaagctt cggtccatta ataactataa ccccaaagat
     4021 tttgactgga atctgaaaca tggccgggtt ttcatcatta agagctactc tgaggacgat
     4081 attcaccgtt ccattaagta taatatttgg tgcagcacag agcatggtaa caagagactg
     4141 gatgctgctt atcgttccat gaacgggaaa ggccccgttt acttactttt cagtgtcaac
     4201 ggcagtggac acttctgtgg cgtggcagaa atgaaatctg ctgtggacta caacacatgt
     4261 gcaggtgtgt ggtcccagga caaatggaag ggtcgttttg atgtcaggtg gatttttgtg
     4321 aaggacgttc ccaatagcca actgcgacac attcgcctag agaacaacga gaataaacca
     4381 gtgaccaact ctagggacac tcaggaagtg cctctggaaa aggctaagca ggtgttgaaa
     4441 attatagcca gctacaagca caccacttcc atttttgatg acttctcaca ctatgagaaa
     4501 cgccaagagg aagaagaaag tgttaaaaag gaacgtcaag gtcgtgggaa aggcggtagt
     4561 tatccttatg acgtgcctga ctatgcctga ggatcccgcc cctctccctc ccccccccct
     4621 aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt
     4681 tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg
     4741 acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc
     4801 gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt
     4861 tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta
     4921 taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg
     4981 gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag
     5041 gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag
     5101 tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa
     5161 acacgatgat aagcttgcca caaccatggc tgaacaagat ggattgcacg caggttctcc
     5221 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     5281 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     5341 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     5401 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     5461 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     5521 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     5581 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     5641 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     5701 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     5761 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     5821 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     5881 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     5941 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga acgcgtctgg aacaatcaac
     6001 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     6061 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     6121 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     6181 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     6241 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     6301 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     6361 ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg
     6421 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     6481 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     6541 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaatt aattctgcag
     6601 tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga
     6661 ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt
     6721 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     6781 gaggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat
     6841 ctaccacaca caaggctact tccctgatta gcagaactac acaccagggc caggggtcag
     6901 atatccactg acctttggat ggtgctacaa gctagtacca gttgagccag ataaggtaga
     6961 agaggccaat aaaggagaga acaccagctt gttacaccct gtgagcctgc atgggatgga
     7021 tgacccggag agagaagtgt tagagtggag gtttgacagc cgcctagcat ttcatcacgt
     7081 ggcccgagag ctgcatccgg agtacttcaa gaactgctga tatcgagctt gctacaaggg
     7141 actttccgct ggggactttc cagggaggcg tggcctgggc gggactgggg agtggcgagc
     7201 cctcagatcc tgcatataag cagctgcttt ttgcctgtac tgggtctctc tggttagacc
     7261 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     7321 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     7381 gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc
     7441 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggccttgac
     7501 attgctagcg tttaccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     7561 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     7621 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     7681 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     7741 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     7801 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     7861 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     7921 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     7981 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     8041 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     8101 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     8161 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     8221 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     8281 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     8341 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     8401 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     8461 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     8521 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     8581 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     8641 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     8701 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     8761 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     8821 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     8881 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     8941 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     9001 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     9061 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     9121 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     9181 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     9241 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     9301 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     9361 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     9421 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     9481 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     9541 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     9601 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     9661 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     9721 atcaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     9781 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     9841 tcttatcatg tctggatcaa ctggataact caagctaacc aaaatcatcc caaacttccc
     9901 accccatacc ctattaccac tgccaattac ctgtggtttc atttactcta aacctgtgat
     9961 tcctctgaat tattttcatt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta
    10021 gtagt
//
