LOCUS       V032331                 9461 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032331
VERSION     V032331
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..4482
                     /label="NDUFS1(NM_005006)"
                     /note="NDUFS1(NM_005006)"
                     /gene="NDUFS1"
     CDS             4489..4515
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4519..4545
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4549..4575
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     promoter        4626..4837
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4850..5118
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5151..5549
                     /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    5556..6144
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6218..6451
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6523..6644
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6663..6798
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6836..6852)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6860..6876
                     /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(6884..6914)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6929..6950
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7238..7826)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7997..8857)
                     /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(8858..8962)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9436..9452
                     /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 catgttaagg atacctgtaa gaaaggcctt agtaggcctt
     2341 tctaagtctc ctaaaggatg tgttcgaaca actgccacag cagcaagcaa cttgattgaa
     2401 gtatttgttg atggtcagtc tgtcatggtg gaaccgggaa cgaccgtcct ccaagcttgt
     2461 gagaaggttg gcatgcagat ccctcgattc tgttatcatg aaaggttgtc tgttgctgga
     2521 aactgcagga tgtgccttgt tgaaattgag aaagccccta aggttgtagc tgcttgtgcc
     2581 atgccagtaa tgaagggttg gaatatccta acaaactcag aaaaatccaa aaaagccagg
     2641 gaaggtgtga tggagttctt attagcaaat cacccattgg actgtcctat ttgtgaccag
     2701 ggaggtgaat gtgatctgca ggaccagtcc atgatgtttg gaaatgatag gagccgattt
     2761 ttagagggga agcgtgctgt ggaagacaag aacattgggc cattggtaaa gaccatcatg
     2821 acaagatgta tacagtgtac tcgctgcatc aggtttgcaa gtgagattgc aggagtagat
     2881 gatttgggaa caacaggcag aggaaatgat atgcaagttg gcacatacat tgaaaagatg
     2941 ttcatgtctg aactgtctgg gaatatcatt gatatctgcc ctgtaggtgc cctaacctct
     3001 aagccctatg cctttactgc ccggccttgg gaaacaagaa agacagaatc cattgatgta
     3061 atggatgcgg ttggaagtaa tattgtggtt agcacaagaa ctggagaagt gatgaggatt
     3121 ttgccacgta tgcatgagga catcaatgaa gagtggatct ctgataaaac cagatttgcc
     3181 tatgatgggc taaaacgtca aagacttacc gagccaatgg tcagaaatga aaaagggctt
     3241 ttaacctata cttcttggga ggatgcgctc tctcgcgtag ctggaatgtt gcagagtttt
     3301 caaggcaaag atgtggcagc aattgcaggt ggcttggtgg atgctgaagc cctggtagct
     3361 ctcaaagatt tgcttaatag agtggactct gacaccttat gcactgaaga ggtcttcccc
     3421 actgcaggag ctggcacaga tttgcgttcc aattatcttc ttaatactac aattgctggt
     3481 gtggaagagg cagatgttgt tcttctggtt ggtacaaacc cacgttttga ggcaccactg
     3541 tttaatgcta gaattcgaaa gagctggctg cataatgact taaaagtggc ccttataggc
     3601 agtccagtgg acctcactta cacatatgac cacctgggag actcccccaa aattcttcaa
     3661 gacattgctt cgggaagcca tccatttagc caggtcctaa aggaagctaa aaaaccaatg
     3721 gtggttttag gcagttctgc actccaaaga aatgatggag cagcaattct tgcagctgtt
     3781 tctagcattg cacaaaagat tcggatgact agtggtgtta ctggtgattg gaaagttatg
     3841 aatatccttc ataggattgc aagtcaagta gctgctttgg accttggcta taagcctggg
     3901 gtggaagcaa ttcggaagaa ccctcccaag gtgctgtttc tcctgggagc agatggaggt
     3961 tgtatcacac gacaggattt gccaaaggat tgtttcatta tttatcaagg acatcatggt
     4021 gatgttgggg ctcccatagc tgatgttatt ctcccaggag ctgcttacac agagaagtct
     4081 gctacatatg tcaacactga gggtagagct cagcagacta aggtagcagt gacacctcct
     4141 ggcttggcaa gagaagactg gaaaattata agagcactct ctgagattgc tggaatgact
     4201 cttccatatg atactctgga tcaagtaagg aacagattgg aagaagtctc tcctaatctt
     4261 gttcgatatg atgatattga aggggctaat tacttccagc aagcaaatga gctctcaaag
     4321 ctagtgaacc agcagcttct tgctgaccca cttgttccac ctcagctaac tataaaagac
     4381 ttctacatga cagattcaat tagcagagcc tcacagacaa tggccaaatg tgtcaaagct
     4441 gtcacagagg gtgcccaggc agtagaggaa ccatccatat gcctcgagta cccatacgac
     4501 gtacctgatt atgcaactta tccttatgat gtacctgatt atgctagcta tccttatgat
     4561 gtcccagact acgcataggg atccgcggcc gcgaaggatc tgcgatcgct ccggtgcccg
     4621 tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa
     4681 ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg
     4741 gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa
     4801 cgttcttttt cgcaacgggt ttgccgccag aacacagctg aagcttcgag gggctcgcat
     4861 ctctccttca cgcgcccgcc gccctacctg aggccgccat ccacgccggt tgagtcgcgt
     4921 tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa
     4981 gctcaggtcg agaccgggcc tttgtccggc gctcccttgg agcctaccta gactcagccg
     5041 gctctccacg ctttgcctga ccctgcttgc tcaactctac gtctttgttt cgttttctgt
     5101 tctgcgccgt tacagatcca agctgtgacc ggcgcctacg ctaggccacc atggccaagc
     5161 ctttgtctca agaagaatcc accctcattg aaagagcaac ggctacaatc aacagcatcc
     5221 ccatctctga agactacagc gtcgccagcg cagctctctc tagcgacggc cgcatcttca
     5281 ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc agaactcgtg gtgctgggca
     5341 ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga aatgagaaca
     5401 ggggcatctt gagcccctgc ggacggtgcc gacaggtgct tctcgatctg catcctggga
     5461 tcaaagccat agtgaaggac agtgatggac agccgacggc agttgggatt cgtgaattgc
     5521 tgccctctgg ttatgtgtgg gagggctaag tcgacaatca acctctggat tacaaaattt
     5581 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     5641 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     5701 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     5761 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     5821 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     5881 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     5941 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
     6001 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     6061 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     6121 tctccctttg ggccgcctcc ccgcctggta cctttaagac caatgactta caaggcagct
     6181 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     6241 cgaaaataag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
     6301 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     6361 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     6421 cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag
     6481 tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct
     6541 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     6601 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag
     6661 ctatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     6721 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     6781 gaggcttttt tggaggccta gacttttgca gagacggccc aaattcgtaa tcatggtcat
     6841 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     6901 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     6961 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     7021 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     7081 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     7141 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     7201 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     7261 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     7321 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     7381 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     7441 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     7501 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     7561 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     7621 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     7681 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     7741 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     7801 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     7861 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     7921 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     7981 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     8041 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     8101 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     8161 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     8221 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     8281 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     8341 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     8401 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     8461 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     8521 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     8581 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     8641 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     8701 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     8761 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     8821 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     8881 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     8941 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     9001 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     9061 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     9121 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     9181 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     9241 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt
     9301 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     9361 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     9421 cccagtcacg acgttgtaaa acgacggcca gtgccaagct g
//
