LOCUS       V043684                 9941 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043684
VERSION     V043684
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..3018
                     /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"
     CDS             3055..4866
                     /label="Nfe2l2(NM_001399173)"
                     /note="Nfe2l2(NM_001399173)"
                     /gene="Nfe2l2"
     promoter        4911..5122
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5135..5403
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5430..6029
                     /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    6036..6624
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6698..6931
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    7003..7124
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7143..7278
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7316..7332)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7340..7356
                     /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(7364..7394)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7409..7430
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7718..8306)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8477..9337)
                     /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(9338..9442)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9916..9932
                     /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 catggtgagc aagggcgagg agctgttcac cggggtggtg
     2341 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     2401 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     2461 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     2521 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     2581 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     2641 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     2701 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     2761 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     2821 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     2881 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     2941 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     3001 atggacgagc tgtacaaggg cggcggaggg agtggaggag ggggaagtgg atccatgatg
     3061 gacttggaat tgccaccgcc aggactacag tcccagcagg acatggattt gattgacatc
     3121 ctttggaggc aagacataga tcttggggta agtcgagaag tgtttgactt tagtcagcga
     3181 cagaaggatt atgagctgga aaaacagaaa aaactcgaaa aggagagaca agagcaactc
     3241 cagaaggaac aggagaaggc ctttttcgct cagttacaac tggatgaaga gaccggagaa
     3301 ttcctcccaa ttcagccagc ccagcacatc cagacagaca ccagtggatc tgtcagctac
     3361 tcccaggttg cccacattcc caaacaagat gccttgtact ttgaagactg tatgcagctt
     3421 ttggcagaga cattcccatt tgtagatgac catgaggttt cttcacctac atttcagtcg
     3481 cttgccctgg atattcccag ccacgttgag agctcagtct tcaccacccc tgatcaggct
     3541 cagtcactcg atagctctct ggagacggcc atgactgatt taagcagcat acagcaggac
     3601 atggagcaag tttggcagga gctattttcc attcccgagt tacagtgtct taatacggaa
     3661 aacaagcagc aggctgagac taccactgtc cccagcccag aggccacact gacagagatg
     3721 gacagcaatt accattttta ctcatcgatc ccctcactgg aaaaagaagt ggacagctgt
     3781 agtccacatt tccttcatgg ttttgaggat tcttttagta gcatcctctc cacggatgat
     3841 gccagccagc tgaactcctt agactcaaat cccaccttga acacagattt cggtgatgaa
     3901 ttttactctg ctttcctagc agagcccagt ggcggtggca gcatgccttc ctctgctgcc
     3961 attagtcagt cgctctctga acttctgggc gggcccattg agggctgtga tctgtccctg
     4021 tgtaaagctt tcaaccagaa gcacactgaa ggcacggtgg agttcaatga ctctgactcc
     4081 ggcatttcgc tgaacacaag tcccagccga gcatccccag agcactctgt ggagtcttcc
     4141 atttacggag acccaccgcc tgggttcagt gactcggaaa tggaagagct agacagtgcc
     4201 cctggaagtg tcaaacagaa tggacctaaa gcacagccaa cacattcttc tggggataca
     4261 gtacagcctc tgtcgccagc tcaagggcac agtgctgctg tgcacgaatc ccagtgtgaa
     4321 aatacaacaa aaaaagaagt acctgtgagt cctggtcatc aaaaagtccc attcacaaaa
     4381 gacaaacatt caagccgatt agaggctcat ctcacaagag atgagcttag ggcaaaagct
     4441 ctccatattc cattccctgt tgaaaaaatc attaatctcc ctgttgatga cttcaatgaa
     4501 atgatgtcca aggagcaatt caacgaagct cagcttgcat taattcgaga tatacgcagg
     4561 agagggaaga ataaagttgc cgctcagaac tgtaggaaaa ggaagctgga aaacattgta
     4621 gagctggagc aagacttggg ccacttaaaa gacgagagag aaaaactact cagagaaaag
     4681 ggagaaaacg acagaaacct ccatcttctg aaaagaaaac tcagcacttt gtatctggaa
     4741 gtcttcagca tgttacgtga tgaggatggg aaaccttact ctcccagtga gtactctctg
     4801 cagcagacca gagatggcaa cgtgttcctt gttcccaaga gcaagaagcc agatacaaag
     4861 aaaaactagg cggccgcgaa ggatctgcga tcgctccggt gcccgtcagt gggcagagcg
     4921 cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa cgggtgccta
     4981 gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc
     5041 cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa
     5101 cgggtttgcc gccagaacac agctgaagct tcgaggggct cgcatctctc cttcacgcgc
     5161 ccgccgccct acctgaggcc gccatccacg ccggttgagt cgcgttctgc cgcctcccgc
     5221 ctgtggtgcc tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc
     5281 gggcctttgt ccggcgctcc cttggagcct acctagactc agccggctct ccacgctttg
     5341 cctgaccctg cttgctcaac tctacgtctt tgtttcgttt tctgttctgc gccgttacag
     5401 atccaagctg tgaccggcgc ctacgctaga tgaccgagta caagcccacg gtgcgcctcg
     5461 ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc cgccgcgttc gccgactacc
     5521 ccgccacgcg ccacaccgtc gacccggacc gccacatcga gcgggtcacc gagctgcaag
     5581 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     5641 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     5701 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     5761 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
     5821 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     5881 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     5941 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     6001 ggtgcatgac ccgcaagccc ggtgcctgag tcgacaatca acctctggat tacaaaattt
     6061 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     6121 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     6181 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     6241 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     6301 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     6361 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     6421 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
     6481 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     6541 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     6601 tctccctttg ggccgcctcc ccgcctggta cctttaagac caatgactta caaggcagct
     6661 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     6721 cgaaaataag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
     6781 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     6841 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     6901 cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag
     6961 tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct
     7021 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     7081 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag
     7141 ctatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     7201 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     7261 gaggcttttt tggaggccta gacttttgca gagacggccc aaattcgtaa tcatggtcat
     7321 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     7381 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     7441 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     7501 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     7561 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     7621 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     7681 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     7741 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     7801 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     7861 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     7921 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     7981 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     8041 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     8101 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     8161 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     8221 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     8281 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     8341 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     8401 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     8461 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     8521 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     8581 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     8641 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     8701 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     8761 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     8821 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     8881 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     8941 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     9001 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     9061 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     9121 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     9181 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     9241 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     9301 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     9361 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     9421 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     9481 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     9541 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     9601 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     9661 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     9721 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt
     9781 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     9841 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     9901 cccagtcacg acgttgtaaa acgacggcca gtgccaagct g
//
