LOCUS       V032901                 8981 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032901
VERSION     V032901
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             2305..2331
                     /codon_start=1
                     /product="nuclear localization signal (Makkerh et al.,
                     1996)"
                     /transl_table=1
                     /translation="PAAKKKKLD"
                     /label="NLS"
     CDS             2332..3900
                     /label="EARS2(NM_001083614)"
                     /note="EARS2(NM_001083614)"
                     /gene="EARS2"
     promoter        3951..4162
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4175..4443
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4470..5069
                     /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    5076..5664
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5738..5971
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6043..6164
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6183..6318
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6356..6372)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6380..6396
                     /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(6404..6434)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6449..6470
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6758..7346)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7517..8377)
                     /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(8378..8482)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8956..8972
                     /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 catgccagca gcgaagaaaa agaagctgga tatggcggcg
     2341 ctcctgagga gactgctgca gcgcgagagg ccttcggcgg cctctggccg ccccgtagga
     2401 cggcgcgagg ccaacctggg cactgatgcc ggggttgcgg tgcgagtgcg gttcgctccc
     2461 agccccacag gcttcttgca cctgggtggc ctccgcactg ccttgtacaa ctacatcttt
     2521 gctaagaagt accaggggag cttcatcctg aggctagagg acacagatca gactcgcgtt
     2581 gtgcctgggg cagcggagaa tattgaggac atgctggagt gggcaggcat cccgcctgat
     2641 gagagccccc gccggggcgg tcctgctggg ccctaccagc aatctcagcg gttggagctg
     2701 tatgcccagg ccacagaagc gctgctgaag accggagctg cttacccctg tttctgctca
     2761 ccccagcggc tggagctcct gaagaaggag gccttgcgga accaccagac gccccggtat
     2821 gacaatcggt gcaggaacat gagccaggag caggtggccc agaagctggc caaggacccc
     2881 aagcctgcga tccgcttccg cctggagcag gtggtgccag ccttccagga cctggtctat
     2941 ggctggaata ggcatgaagt ggccagcgtg gagggagacc cagtcatcat gaagagcgac
     3001 ggcttcccca cataccacct ggcctgcgtg gtggacgacc accacatggg catcagccac
     3061 gtgctgcgag gctctgagtg gctcgtctcc actgccaagc acctgctcct ctaccaggcc
     3121 ctgggctggc agccacccca cttcgcccac ctgcccctgc tcctcaacag ggatggcagc
     3181 aagctctcca agaggcaagg ggacgttttc ctggagcact ttgctgctga tggcttcctg
     3241 cccgattcct tgttggacat catcaccaac tgtggctcag gttttgcaga gaaccaaatg
     3301 ggcaggaccc tgccggagct gatcacacag ttcaacctga cacaggtcac ctgtcactca
     3361 gccctgctgg acctggagaa gctcccagaa ttcaacagac tgcacctcca gcggctggtg
     3421 agcaatgaga gccagaggcg ccagctggtg gggaagctgc aggtccttgt ggaggaggcc
     3481 tttggttgcc agctgcaaaa cagggatgtc ctcaacccag tctacgtgga gaggatcctc
     3541 ctgctgagac agggtcacat ttgccgcctg caggacttgg tgtccccagt atactcttac
     3601 ctgtggactc gccctgcagt aggtcgagca cagctggacg ccatctcgga gaaggtggat
     3661 gtgattgcca agcgtgtgct ggggcttcta gaaagatcta gtatgagctt aactcaggat
     3721 atgctgaatg gagaactgaa gaagctatca gaaggtctgg aaggcaccaa gtacagtaat
     3781 gtgatgaaac tccttcggat ggccctcagt ggacagcagc aaggacctcc tgtagctgag
     3841 atgatgttgg ccttgggacc aaaggaagta cgggaacgga tccagaaggt ggtttccagc
     3901 tagggatccg cggccgcgaa ggatctgcga tcgctccggt gcccgtcagt gggcagagcg
     3961 cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa cgggtgccta
     4021 gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc
     4081 cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa
     4141 cgggtttgcc gccagaacac agctgaagct tcgaggggct cgcatctctc cttcacgcgc
     4201 ccgccgccct acctgaggcc gccatccacg ccggttgagt cgcgttctgc cgcctcccgc
     4261 ctgtggtgcc tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc
     4321 gggcctttgt ccggcgctcc cttggagcct acctagactc agccggctct ccacgctttg
     4381 cctgaccctg cttgctcaac tctacgtctt tgtttcgttt tctgttctgc gccgttacag
     4441 atccaagctg tgaccggcgc ctacgctaga tgaccgagta caagcccacg gtgcgcctcg
     4501 ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc cgccgcgttc gccgactacc
     4561 ccgccacgcg ccacaccgtc gacccggacc gccacatcga gcgggtcacc gagctgcaag
     4621 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     4681 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     4741 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     4801 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
     4861 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     4921 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     4981 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     5041 ggtgcatgac ccgcaagccc ggtgcctgag tcgacaatca acctctggat tacaaaattt
     5101 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     5161 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     5221 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     5281 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     5341 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     5401 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     5461 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
     5521 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     5581 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     5641 tctccctttg ggccgcctcc ccgcctggta cctttaagac caatgactta caaggcagct
     5701 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     5761 cgaaaataag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
     5821 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     5881 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     5941 cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag
     6001 tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct
     6061 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     6121 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag
     6181 ctatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     6241 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     6301 gaggcttttt tggaggccta gacttttgca gagacggccc aaattcgtaa tcatggtcat
     6361 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     6421 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     6481 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     6541 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     6601 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     6661 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6721 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6781 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6841 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6901 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6961 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     7021 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     7081 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     7141 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     7201 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     7261 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     7321 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     7381 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     7441 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     7501 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     7561 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7621 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7681 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7741 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7801 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7861 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7921 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7981 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     8041 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     8101 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     8161 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     8221 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     8281 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     8341 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     8401 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     8461 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     8521 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     8581 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     8641 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     8701 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     8761 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt
     8821 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     8881 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     8941 cccagtcacg acgttgtaaa acgacggcca gtgccaagct g
//
