LOCUS       V047231                 9247 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047231
VERSION     V047231
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..3876
                     /label="USP22(NM_015276)"
                     /note="USP22(NM_015276)"
                     /gene="USP22"
     CDS             3883..3972
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     promoter        4016..4227
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4240..4508
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4541..5335
                     /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    5342..5930
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6004..6237
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6309..6430
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6449..6584
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6622..6638)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6646..6662
                     /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(6670..6700)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6715..6736
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7024..7612)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7783..8643)
                     /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(8644..8748)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9222..9238
                     /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 catggtgtcc cggccagagc ccgagggcga ggccatggac
     2341 gccgagctgg cggtagcgcc gccgggctgc tcgcacctgg gcagcttcaa ggtggacaac
     2401 tggaagcaga acctgcgggc catctaccag tgcttcgtgt ggagcggcac ggctgaggcc
     2461 cgcaagcgca aggccaagtc ctgtatctgc catgtctgtg gcgtccacct caacaggctg
     2521 cattcctgcc tctactgtgt cttcttcggc tgtttcacaa agaagcatat tcacgagcat
     2581 gcgaaggcga agcggcacaa cctggccatt gatctgatgt acggaggcat ctactgtttt
     2641 ctgtgccagg actacatcta tgacaaagac atggaaataa tcgccaagga ggagcagcga
     2701 aaagcttgga aaatgcaagg cgttggagag aagttttcaa cttgggaacc aaccaaacgg
     2761 gagcttgaac tgctgaagca caacccgaaa aggagaaaga tcacctcgaa ctgcaccata
     2821 ggtctgcgtg ggctgatcaa ccttgggaac acatgcttca tgaactgcat cgtgcaggcc
     2881 ctgacccaca cgccacttct gcgggacttc ttcctgtctg acaggcaccg ctgtgagatg
     2941 cagagcccca gctcctgtct ggtctgtgag atgtcctcac tgtttcagga gttttactct
     3001 ggacaccggt cccctcacat cccgtataag ttgctgcacc tggtgtggac ccacgcgagg
     3061 cacctagcag gctacgagca gcaggacgcc cacgagttcc tcatcgcggc cctggacgtg
     3121 ctccaccgac actgcaaagg tgatgacaat gggaagaagg ccaacaaccc caaccactgc
     3181 aactgcatca tagaccagat cttcacaggc gggttgcagt cagacgtcac ctgccaagtc
     3241 tgccatggag tctccaccac catcgacccc ttctgggaca tcagcttgga tctccccggc
     3301 tcttccaccc cattctggcc cctgagccca gggagcgagg gcaacgtggt aaacggggaa
     3361 agccacgtgt cgggaaccac cacgctcacg gactgcctgc gacgattcac cagaccagag
     3421 cacttgggca gcagcgccaa gatcaagtgc agcggttgcc atagctacca ggagtccaca
     3481 aagcagctca ctatgaagaa actgcccatc gtagcctgtt ttcatctcaa acgatttgaa
     3541 cactcagcca agctgcggcg gaagatcacc acgtatgtgt ccttccccct ggagctggac
     3601 atgacccctt tcatggcctc cagcaaagag agcaggatga atggacagta ccagcagccc
     3661 acggacagtc tcaacaatga caacaagtat tccctgtttg ctgttgttaa ccatcaaggg
     3721 accttggaga gtggccacta caccagcttt atccggcagc acaaagacca gtggttcaag
     3781 tgtgacgatg ccatcatcac caaggccagc atcaaggacg tcctggacag cgaagggtac
     3841 ttgctgttct atcacaaaca gttcctggaa tacgagggat ccgaacaaaa actaatatca
     3901 gaggaagatt tggaacagaa actaatcagt gaagaagatt tagagcaaaa gttaatttcc
     3961 gaggaggact tatagcggcc gcgaaggatc tgcgatcgct ccggtgcccg tcagtgggca
     4021 gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaacgggt
     4081 gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt
     4141 tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt
     4201 cgcaacgggt ttgccgccag aacacagctg aagcttcgag gggctcgcat ctctccttca
     4261 cgcgcccgcc gccctacctg aggccgccat ccacgccggt tgagtcgcgt tctgccgcct
     4321 cccgcctgtg gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg
     4381 agaccgggcc tttgtccggc gctcccttgg agcctaccta gactcagccg gctctccacg
     4441 ctttgcctga ccctgcttgc tcaactctac gtctttgttt cgttttctgt tctgcgccgt
     4501 tacagatcca agctgtgacc ggcgcctacg ctaggccacc atggctgaac aagatggatt
     4561 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     4621 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     4681 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     4741 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     4801 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     4861 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     4921 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     4981 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     5041 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     5101 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     5161 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     5221 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     5281 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagtcga
     5341 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     5401 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     5461 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     5521 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     5581 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     5641 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     5701 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     5761 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     5821 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     5881 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt
     5941 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
     6001 gactggaagg gctaattcac tcccaacgaa aataagatct gctttttgct tgtactgggt
     6061 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     6121 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     6181 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta
     6241 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga
     6301 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     6361 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     6421 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca gttccgccca
     6481 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     6541 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga
     6601 cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     6661 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     6721 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     6781 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6841 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     6901 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     6961 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     7021 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     7081 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     7141 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     7201 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     7261 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     7321 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     7381 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     7441 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     7501 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     7561 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     7621 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     7681 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     7741 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     7801 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     7861 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     7921 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     7981 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     8041 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     8101 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     8161 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     8221 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     8281 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     8341 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     8401 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     8461 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     8521 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     8581 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     8641 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     8701 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     8761 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     8821 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     8881 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     8941 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     9001 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     9061 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     9121 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     9181 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc
     9241 caagctg
//
