LOCUS       V042355                 9358 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042355
VERSION     V042355
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             234..414
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    458..583
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1076..1309
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1493..1537
                     /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    1804..1920
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2010..2213
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2302..4212
                     /label="PRMT5(NM_006109)"
                     /note="PRMT5(NM_006109)"
                     /gene="PRMT5"
     CDS             4219..4284
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4328..4539
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4552..4820
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4847..5446
                     /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    5453..6041
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6115..6348
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6420..6541
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6560..6695
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6733..6749)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6757..6773
                     /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(6781..6811)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6826..6847
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7135..7723)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7894..8754)
                     /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(8755..8859)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9333..9349
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt
     1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag
     2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct
     2281 agagctagcg aattcgccac catggcggcg atggcggtcg ggggtgctgg tgggagccgc
     2341 gtgtccagcg ggagggacct gaattgcgtc cccgaaatag ctgacacact aggggctgtg
     2401 gccaagcagg ggtttgattt cctctgcatg cctgtcttcc atccgcgttt caagagggag
     2461 ttcattcagg aacctgctaa gaatcggccc ggtccccaga cacgatcaga cctactgctg
     2521 tcaggaaggg actggaatac gctaattgtg ggaaagcttt ctccatggat tcgtccagac
     2581 tcaaaagtgg agaagattcg caggaactcc gaggcggcca tgttacagga gctgaatttt
     2641 ggtgcatatt tgggtcttcc agctttcctg ctgcccctta atcaggaaga taacaccaac
     2701 ctggccagag ttttgaccaa ccacatccac actggccatc actcttccat gttctggatg
     2761 cgggtaccct tggtggcacc agaggacctg agagatgata taattgagaa tgcaccaact
     2821 acacacacag aggagtacag tggggaggag aaaacgtgga tgtggtggca caacttccgg
     2881 actttgtgtg actatagtaa gaggattgca gtggctcttg aaattggggc tgacctccca
     2941 tctaatcatg tcattgatcg ctggcttggg gagcccatca aagcagccat tctccccact
     3001 agcattttcc tgaccaataa gaagggattt cctgttcttt ctaagatgca ccagaggctc
     3061 atcttccggc tcctcaagtt ggaggtgcag ttcatcatca caggcaccaa ccaccactca
     3121 gagaaggagt tctgctccta cctccaatac ctggaatact taagccagaa ccgtcctcca
     3181 cctaatgcct atgaactctt tgccaagggc tatgaagact atctgcagtc cccgcttcag
     3241 ccactgatgg acaatctgga atctcagaca tatgaagtgt ttgaaaagga ccccatcaaa
     3301 tactctcagt accagcaggc catctataaa tgtctgctag accgagtacc agaagaggag
     3361 aaggatacca atgtccaggt actgatggtg ctgggagcag gacggggacc cctggtgaac
     3421 gcttccctgc gggcagccaa gcaggccgac cggcggataa agctgtatgc tgtggagaaa
     3481 aacccaaatg ccgtggtgac gctagagaac tggcagtttg aagaatgggg aagccaagtg
     3541 accgtagtct catcagacat gagggaatgg gtggctccag agaaagcaga catcattgtc
     3601 agtgagcttc tgggctcatt tgctgacaat gaattgtcgc ctgagtgcct ggatggagcc
     3661 cagcacttcc taaaagatga tggtgtgagc atccccgggg agtacacttc ctttctggct
     3721 cccatctctt cctccaagct gtacaatgag gtccgagcct gtagggagaa ggaccgtgac
     3781 cctgaggccc agtttgagat gccttatgtg gtacggctgc acaacttcca ccagctctct
     3841 gcaccccagc cctgtttcac cttcagccat cccaacagag atcctatgat tgacaacaac
     3901 cgctattgca ccttggaatt tcctgtggag gtgaacacag tactacatgg ctttgccggc
     3961 tactttgaga ctgtgcttta tcaggacatc actctgagta tccgtccaga gactcactct
     4021 cctgggatgt tctcatggtt tcccatcctc ttccctatta agcagcccat aacggtacgt
     4081 gaaggccaaa ccatctgtgt gcgtttctgg cgatgcagca attccaagaa ggtgtggtat
     4141 gagtgggctg tgacagcacc agtctgttct gctattcata accccacagg ccgctcatat
     4201 accattggcc tcggatccga ctacaaagac catgacggtg attataaaga tcatgacatc
     4261 gactacaagg atgacgatga caagtagcgg ccgcgaagga tctgcgatcg ctccggtgcc
     4321 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc
     4381 aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac
     4441 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg
     4501 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc
     4561 atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc
     4621 gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta
     4681 aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc
     4741 cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct
     4801 gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagatga ccgagtacaa
     4861 gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca ccctcgccgc
     4921 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc acatcgagcg
     4981 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     5041 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     5101 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     5161 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     5221 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     5281 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc
     5341 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     5401 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgagtcg acaatcaacc
     5461 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     5521 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     5581 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     5641 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     5701 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     5761 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     5821 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     5881 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     5941 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     6001 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggtacct ttaagaccaa
     6061 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     6121 ggctaattca ctcccaacga aaataagatc tgctttttgc ttgtactggg tctctctggt
     6181 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     6241 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     6301 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat
     6361 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga
     6421 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     6481 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     6541 atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc attctccgcc
     6601 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     6661 attccagaag tagtgaggag gcttttttgg aggcctagac ttttgcagag acggcccaaa
     6721 ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     6781 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     6841 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     6901 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     6961 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     7021 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     7081 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     7141 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     7201 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     7261 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     7321 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     7381 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     7441 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     7501 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     7561 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     7621 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     7681 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     7741 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     7801 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     7861 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     7921 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     7981 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     8041 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     8101 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     8161 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     8221 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     8281 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     8341 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     8401 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     8461 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     8521 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     8581 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     8641 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     8701 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     8761 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     8821 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     8881 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     8941 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     9001 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     9061 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     9121 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     9181 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     9241 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     9301 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctg
//
