LOCUS       V041187                 9463 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041187
VERSION     V041187
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..4092
                     /label="Nfe2l2(NM_010902)"
                     /note="Nfe2l2(NM_010902)"
                     /gene="Nfe2l2"
     CDS             4099..4188
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     promoter        4232..4443
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4456..4724
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4757..5551
                     /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    5558..6146
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6220..6453
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6525..6646
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6665..6800
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6838..6854)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6862..6878
                     /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(6886..6916)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6931..6952
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7240..7828)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7999..8859)
                     /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(8860..8964)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9438..9454
                     /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 catgatggac ttggagttgc caccgccagg actacagtcc
     2341 cagcaggaca tggatttgat tgacatcctt tggaggcaag acatagatct tggagtaagt
     2401 cgagaagtgt ttgactttag tcagcgacag aaggactatg agctggaaaa acagaaaaaa
     2461 ctcgaaaagg aaagacaaga gcaactccag aaggaacagg agaaggcctt ttttgctcag
     2521 tttcaactgg atgaagaaac aggagaattc ctcccaattc agccggccca gcacatccag
     2581 acagacacca gtggatccgc cagctactcc caggttgccc acattcccaa acaagatgcc
     2641 ttgtactttg aagactgtat gcagcttttg gcagagacat tcccatttgt agatgaccat
     2701 gagtcgcttg ccctggatat ccccagccac gctgaaagtt cagtcttcac tgcccctcat
     2761 caggcccagt ccctcaatag ctctctggag gcagccatga ctgatttaag cagcatagag
     2821 caggacatgg agcaagtttg gcaggagcta ttttccattc ccgaattaca gtgtcttaat
     2881 accgaaaaca agcagctggc tgatactacc gctgttccca gcccagaagc cacactgaca
     2941 gaaatggaca gcaattacca tttttactca tcgatctcct cgctggaaaa agaagtgggc
     3001 aactgtggtc cacatttcct tcatggtttt gaggattctt tcagcagcat cctctccact
     3061 gatgatgcca gccagctgac ctccttagac tcaaatccca ccttaaacac agattttggc
     3121 gatgaatttt attctgcttt catagcagag cccagtgacg gtggcagcat gccttcctcc
     3181 gctgccatca gtcagtcact ctctgaactc ctggacggga ctattgaagg ctgtgacctg
     3241 tcactgtgta aagctttcaa cccgaagcac gctgaaggca caatggaatt caatgactct
     3301 gactctggca tttcactgaa cacgagtccc agccgagcgt ccccagagca ctccgtggag
     3361 tcttccattt acggagaccc accgcctggg ttcagtgact cggaaatgga ggagctagat
     3421 agtgcccctg gaagtgtcaa acagaacggc cctaaagcac agccagcaca ttctcctgga
     3481 gacacagtac agcctctgtc accagctcaa gggcacagtg ctcctatgcg tgaatcccaa
     3541 tgtgaaaata caacaaaaaa agaagttccc gtgagtcctg gtcatcaaaa agccccattc
     3601 acaaaagaca aacattcaag ccgcttagag gctcatctca cacgagatga gcttagggca
     3661 aaagctctcc atattccatt ccctgtcgaa aaaatcatta acctccctgt tgatgacttc
     3721 aatgaaatga tgtccaagga gcaattcaat gaagctcagc tcgcattgat ccgagatata
     3781 cgcaggagag gtaagaataa agtcgccgcc cagaactgta ggaaaaggaa gctggagaac
     3841 attgtcgagc tggagcaaga cttgggccac ttaaaagacg agagagaaaa actactcaga
     3901 gaaaagggag aaaacgacag aaacctccat ctactgaaaa ggcggctcag caccttgtat
     3961 cttgaagtct tcagcatgtt acgtgatgag gatggaaagc cttactctcc cagtgaatac
     4021 tctctgcagc aaaccagaga tggcaatgtg ttccttgttc ccaaaagcaa gaagccagat
     4081 acaaagaaaa acggatccga acaaaaacta atatcagagg aagatttgga acagaaacta
     4141 atcagtgaag aagatttaga gcaaaagtta atttccgagg aggacttata gcggccgcga
     4201 aggatctgcg atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc
     4261 gagaagttgg ggggaggggt cggcaattga acgggtgcct agagaaggtg gcgcggggta
     4321 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg
     4381 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca
     4441 cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc
     4501 cgccatccac gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact
     4561 gcgtccgccg tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc
     4621 ccttggagcc tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa
     4681 ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca gatccaagct gtgaccggcg
     4741 cctacgctag gccaccatgg ctgaacaaga tggattgcac gcaggttctc cggccgcttg
     4801 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     4861 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     4921 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     4981 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     5041 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     5101 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     5161 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     5221 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     5281 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     5341 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     5401 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     5461 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     5521 cttctatcgc cttcttgacg agttcttctg agtcgacaat caacctctgg attacaaaat
     5581 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     5641 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     5701 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     5761 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     5821 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     5881 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     5941 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     6001 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     6061 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     6121 gatctccctt tgggccgcct ccccgcctgg tacctttaag accaatgact tacaaggcag
     6181 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc
     6241 aacgaaaata agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag
     6301 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     6361 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     6421 gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc
     6481 agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag
     6541 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     6601 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct
     6661 agctatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     6721 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     6781 aggaggcttt tttggaggcc tagacttttg cagagacggc ccaaattcgt aatcatggtc
     6841 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     6901 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     6961 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     7021 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     7081 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     7141 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     7201 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     7261 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     7321 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     7381 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     7441 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     7501 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     7561 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     7621 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     7681 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     7741 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     7801 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     7861 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     7921 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     7981 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     8041 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     8101 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     8161 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     8221 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     8281 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     8341 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     8401 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     8461 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     8521 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     8581 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     8641 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     8701 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     8761 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     8821 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     8881 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     8941 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     9001 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     9061 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     9121 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     9181 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     9241 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     9301 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     9361 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     9421 ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctg
//
