LOCUS       V038345                 9416 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038345
VERSION     V038345
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..3717
                     /label="Mlkl(NM_001310613)"
                     /note="Mlkl(NM_001310613)"
                     /gene="Mlkl"
     CDS             3724..3750
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3754..3780
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3784..3810
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3820..4536
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK*"
                     /label="EGFP"
     promoter        4581..4792
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4805..5073
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5106..5504
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /note="confers resistance to blasticidin"
                     /product="blasticidin S deaminase"
                     /transl_table=1
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG*"
                     /label="BSD"
     misc_feature    5511..6099
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6173..6406
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6478..6599
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6618..6753
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6791..6807)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6815..6831
                     /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(6839..6869)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6884..6905
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7193..7781)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7952..8812)
                     /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(8813..8917)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9391..9407
                     /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 catggataaa ttgggacaga tcatcaagtt aggccagctc
     2341 atctatgaac agtgtgaaaa gatgaaatac tgccggaaac aatgccagcg tctaggaaac
     2401 cgtgtgcacg gcctgctaca gcctctccag agactccagg cccaaggaaa gaagaacctg
     2461 cccgatgaca ttactgctgc cctgggccgt tttgatgaag tcctgaagga ggctaaccag
     2521 cagatagaaa agttcagcaa gaagtcccat atttggaagt ttgtgagtgt gggcaatgat
     2581 aagatcctct tccatgaagt gaatgagaag ctgagagacg tctgggagga gctgttgctg
     2641 ctgcttcagg tttatcattg gaataccgtt tcagatgtca gccagccagc atcctggcag
     2701 caggaagatc gacaggatgc agaggaagac ggaaatgaaa atatgaaagt tatcctgatg
     2761 cagttgcaaa ttagcgtgga agaaatcaac aaaaccctga agcaatgctc actaaaaccc
     2821 acacaggaga tcccacaaga tctccaaatc aaggagattc caaaggaaca tcttggacct
     2881 ccgtggacca aactgaagac aagtaaaatg agcaccattt atagaggaga gtatcacaga
     2941 tctccagtta ccatcaaagt attcaacaac ccccaggccg aaagtgttgg aatagtgagg
     3001 ttcactttca atgacgagat caaaaccatg aagaaattcg attctcccaa catcttgcgt
     3061 atatttggga tttgcattga tcaaacagtg aagccccctg agttctccat tgtcatggag
     3121 tactgtgaac ttggaaccct gagggaactg ctggatagag aaaaagacct cacaatgagt
     3181 gtgcgcagcc tcctagtcct gagggcagcc agaggcttat acaggctaca ccattcggaa
     3241 acactccaca gaaacatcag cagctccagt ttcctcgtag ccggaggcta ccaagtaaag
     3301 cttgcaggat ttgagttaag caaaacacag aattccatca gccggacagc aaagagcact
     3361 aaagcagaga gatccagttc aacgatatat gtctcccctg agagactgaa aaatccattt
     3421 tgcctttatg acataaaagc tgaaatatat agctttggaa ttgtactctg ggaaattgcc
     3481 actggaaaga tcccatttga aggctgtgat tctaagaaga tccgggagct ggtggctgag
     3541 gacaagaagc aggaaccagt gggtcaggat tgccctgagt tgttgcggga aatcattaat
     3601 gagtgtcgtg cccatgagcc ctcccaacgg ccctctgtgg acggtaggag tctttctggc
     3661 agagaacgaa tcttggagag actgtctgcg gttgaagaat ccacggacaa gaaggtggga
     3721 tcctacccat acgacgtacc tgattatgca acttatcctt atgatgtacc tgattatgct
     3781 agctatcctt atgatgtccc agactacgca ttaccggtcg tgagcaaggg cgaggagctg
     3841 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     3901 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     3961 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     4021 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     4081 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     4141 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     4201 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     4261 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     4321 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     4381 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     4441 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     4501 gggatcactc tcggcatgga cgagctgtac aagtaataag cggccgcgaa ggatctgcga
     4561 tcgctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
     4621 gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg cgcggggtaa actgggaaag
     4681 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
     4741 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac agctgaagct
     4801 tcgaggggct cgcatctctc cttcacgcgc ccgccgccct acctgaggcc gccatccacg
     4861 ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc tcctgaactg cgtccgccgt
     4921 ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt ccggcgctcc cttggagcct
     4981 acctagactc agccggctct ccacgctttg cctgaccctg cttgctcaac tctacgtctt
     5041 tgtttcgttt tctgttctgc gccgttacag atccaagctg tgaccggcgc ctacgctagg
     5101 ccaccatggc caagcctttg tctcaagaag aatccaccct cattgaaaga gcaacggcta
     5161 caatcaacag catccccatc tctgaagact acagcgtcgc cagcgcagct ctctctagcg
     5221 acggccgcat cttcactggt gtcaatgtat atcattttac tgggggacct tgtgcagaac
     5281 tcgtggtgct gggcactgct gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga
     5341 tcggaaatga gaacaggggc atcttgagcc cctgcggacg gtgccgacag gtgcttctcg
     5401 atctgcatcc tgggatcaaa gccatagtga aggacagtga tggacagccg acggcagttg
     5461 ggattcgtga attgctgccc tctggttatg tgtgggaggg ctaagtcgac aatcaacctc
     5521 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     5581 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     5641 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     5701 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     5761 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     5821 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     5881 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg
     5941 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     6001 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     6061 ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggtaccttt aagaccaatg
     6121 acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg
     6181 ctaattcact cccaacgaaa ataagatctg ctttttgctt gtactgggtc tctctggtta
     6241 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     6301 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     6361 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt
     6421 catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac
     6481 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     6541 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     6601 catgtctggc tctagctatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     6661 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     6721 tccagaagta gtgaggaggc ttttttggag gcctagactt ttgcagagac ggcccaaatt
     6781 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     6841 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     6901 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     6961 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     7021 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     7081 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     7141 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     7201 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     7261 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     7321 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     7381 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     7441 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     7501 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     7561 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     7621 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     7681 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     7741 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     7801 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     7861 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     7921 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     7981 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     8041 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     8101 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     8161 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     8221 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     8281 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     8341 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     8401 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     8461 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     8521 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     8581 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     8641 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     8701 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     8761 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     8821 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     8881 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     8941 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     9001 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     9061 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     9121 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     9181 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     9241 gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     9301 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     9361 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctg
//
