LOCUS       V042181                 9323 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042181
VERSION     V042181
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..3426
                     /label="Prmt6(NM_001393858)"
                     /note="Prmt6(NM_001393858)"
                     /gene="Prmt6"
     CDS             3433..3459
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3463..3489
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3493..3519
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3526..4245
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK*"
                     /label="EGFP"
     promoter        4293..4504
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4517..4785
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4812..5411
                     /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    5418..6006
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6080..6313
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6385..6506
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6525..6660
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6698..6714)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6722..6738
                     /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(6746..6776)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6791..6812
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7100..7688)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7859..8719)
                     /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(8720..8824)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9298..9314
                     /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 catgtcgctg agcaagaaaa gaaagcttga gtcgggggtc
     2341 ggcggcgcag gaggggaggg agctgaggag gaaaatggcg gggagcagga ggcagccccg
     2401 ccacgacccc ggaggaccaa gcgcgagcgg gaccagctgt actacgagtg ctactccgac
     2461 gtatcggtcc acgaggagat gatcgccgac cgcgtccgca ccgacgccta ccgcctaggc
     2521 atcctgagga actgggccgc tctgcgaggc aagaccgtgc tggacgtggg cgcaggcacc
     2581 ggcattctta gcatcttctg cgcccaggcc ggggcccggc gcgtatacgc tgtggaggcc
     2641 agcgccatct ggcaacaggc ccaggaggtg gtgcggctca acgggttgga ggaccgcgtg
     2701 cacatcctgc caggcccggt ggagacggtg gagctgccgg agcaagtgga cgccatcgta
     2761 agcgagtgga tgggctacgg actactgcac gagtccatgc tgagctccgt gctccacgcg
     2821 cggaccaaat ggctgaagga gggcggtctc ctcctgccag attccgctga gctcttcgtg
     2881 gccccgatta gcgaccagat gctggaatgg cgcttgggtt tctggagcca agtgaagcag
     2941 cactatggcg tggatatgag ctgcatggaa agcttcgcca cgcgctgcct catgggccat
     3001 tcggagatcg tggtgcaggg tctgtccgga gaggacgtgc tggcccggcc acagcgcttt
     3061 gcccagctcg agctggcccg cgctggcctg gagcaggagc tggaggccgg tgtgggcggg
     3121 cgcttccgct gcagctgcta tggctccgcg cctctacatg gttttgccat ctggtttcag
     3181 gtaacctttc ctggagggga ctcggagaaa cctctggtgc tgtccacctc acctttccac
     3241 ccggccacgc actggaagca ggcgctcctc tacttgaatg agccggtgcc ggtggaacaa
     3301 gatacagaca tttctggaga gatcaccctg ctgccctccc gggacaaccc ccggcgtctg
     3361 cgcgtacttc tgcgctacaa agtgggggac catgaggaaa agaccaaaga ctttgccatg
     3421 gaggacggat cctacccata cgacgtacct gattatgcaa cttatcctta tgatgtacct
     3481 gattatgcta gctatcctta tgatgtccca gactacgcag ccaccatggt gagcaagggc
     3541 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     3601 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     3661 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     3721 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     3781 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     3841 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     3901 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     3961 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     4021 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     4081 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     4141 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     4201 accgccgccg ggatcactct cggcatggac gagctgtaca agtagggatc cgcggccgcg
     4261 aaggatctgc gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc
     4321 cgagaagttg gggggagggg tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt
     4381 aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc
     4441 gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac
     4501 acagctgaag cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg
     4561 ccgccatcca cgccggttga gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac
     4621 tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga ccgggccttt gtccggcgct
     4681 cccttggagc ctacctagac tcagccggct ctccacgctt tgcctgaccc tgcttgctca
     4741 actctacgtc tttgtttcgt tttctgttct gcgccgttac agatccaagc tgtgaccggc
     4801 gcctacgcta gatgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc
     4861 cccgggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg
     4921 tcgacccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg
     4981 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga
     5041 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg
     5101 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc
     5161 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca
     5221 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc
     5281 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca
     5341 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc
     5401 ccggtgcctg agtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     5461 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     5521 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     5581 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     5641 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     5701 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     5761 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     5821 cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     5881 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     5941 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     6001 ccccgcctgg tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt
     6061 ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaaaata agatctgctt
     6121 tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     6181 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     6241 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     6301 aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag
     6361 aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa
     6421 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     6481 ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc
     6541 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     6601 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     6661 tagacttttg cagagacggc ccaaattcgt aatcatggtc atagctgttt cctgtgtgaa
     6721 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     6781 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     6841 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     6901 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     6961 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     7021 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     7081 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     7141 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     7201 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     7261 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     7321 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     7381 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     7441 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     7501 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     7561 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     7621 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     7681 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     7741 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     7801 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     7861 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     7921 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     7981 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     8041 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     8101 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     8161 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     8221 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     8281 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     8341 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     8401 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     8461 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     8521 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     8581 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     8641 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     8701 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     8761 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     8821 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     8881 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     8941 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     9001 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     9061 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     9121 cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca
     9181 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     9241 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
     9301 aaacgacggc cagtgccaag ctg
//
