LOCUS       V032836                 8951 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032836
VERSION     V032836
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..3870
                     /label="EARS2(NM_001083614)"
                     /note="EARS2(NM_001083614)"
                     /gene="EARS2"
     promoter        3921..4132
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4145..4413
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4440..5039
                     /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    5046..5634
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5708..5941
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6013..6134
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6153..6288
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6326..6342)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6350..6366
                     /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(6374..6404)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6419..6440
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6728..7316)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7487..8347)
                     /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(8348..8452)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8926..8942
                     /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 catggcggcg ctcctgagga gactgctgca gcgcgagagg
     2341 ccttcggcgg cctctggccg ccccgtagga cggcgcgagg ccaacctggg cactgatgcc
     2401 ggggttgcgg tgcgagtgcg gttcgctccc agccccacag gcttcttgca cctgggtggc
     2461 ctccgcactg ccttgtacaa ctacatcttt gctaagaagt accaggggag cttcatcctg
     2521 aggctagagg acacagatca gactcgcgtt gtgcctgggg cagcggagaa tattgaggac
     2581 atgctggagt gggcaggcat cccgcctgat gagagccccc gccggggcgg tcctgctggg
     2641 ccctaccagc aatctcagcg gttggagctg tatgcccagg ccacagaagc gctgctgaag
     2701 accggagctg cttacccctg tttctgctca ccccagcggc tggagctcct gaagaaggag
     2761 gccttgcgga accaccagac gccccggtat gacaatcggt gcaggaacat gagccaggag
     2821 caggtggccc agaagctggc caaggacccc aagcctgcga tccgcttccg cctggagcag
     2881 gtggtgccag ccttccagga cctggtctat ggctggaata ggcatgaagt ggccagcgtg
     2941 gagggagacc cagtcatcat gaagagcgac ggcttcccca cataccacct ggcctgcgtg
     3001 gtggacgacc accacatggg catcagccac gtgctgcgag gctctgagtg gctcgtctcc
     3061 actgccaagc acctgctcct ctaccaggcc ctgggctggc agccacccca cttcgcccac
     3121 ctgcccctgc tcctcaacag ggatggcagc aagctctcca agaggcaagg ggacgttttc
     3181 ctggagcact ttgctgctga tggcttcctg cccgattcct tgttggacat catcaccaac
     3241 tgtggctcag gttttgcaga gaaccaaatg ggcaggaccc tgccggagct gatcacacag
     3301 ttcaacctga cacaggtcac ctgtcactca gccctgctgg acctggagaa gctcccagaa
     3361 ttcaacagac tgcacctcca gcggctggtg agcaatgaga gccagaggcg ccagctggtg
     3421 gggaagctgc aggtccttgt ggaggaggcc tttggttgcc agctgcaaaa cagggatgtc
     3481 ctcaacccag tctacgtgga gaggatcctc ctgctgagac agggtcacat ttgccgcctg
     3541 caggacttgg tgtccccagt atactcttac ctgtggactc gccctgcagt aggtcgagca
     3601 cagctggacg ccatctcgga gaaggtggat gtgattgcca agcgtgtgct ggggcttcta
     3661 gaaagatcta gtatgagctt aactcaggat atgctgaatg gagaactgaa gaagctatca
     3721 gaaggtctgg aaggcaccaa gtacagtaat gtgatgaaac tccttcggat ggccctcagt
     3781 ggacagcagc aaggacctcc tgtagctgag atgatgttgg ccttgggacc aaaggaagta
     3841 cgggaacgga tccagaaggt ggtttccagc tagggatccg cggccgcgaa ggatctgcga
     3901 tcgctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
     3961 gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg cgcggggtaa actgggaaag
     4021 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
     4081 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac agctgaagct
     4141 tcgaggggct cgcatctctc cttcacgcgc ccgccgccct acctgaggcc gccatccacg
     4201 ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc tcctgaactg cgtccgccgt
     4261 ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt ccggcgctcc cttggagcct
     4321 acctagactc agccggctct ccacgctttg cctgaccctg cttgctcaac tctacgtctt
     4381 tgtttcgttt tctgttctgc gccgttacag atccaagctg tgaccggcgc ctacgctaga
     4441 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac
     4501 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gacccggacc
     4561 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
     4621 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
     4681 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
     4741 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
     4801 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
     4861 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
     4921 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     4981 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgag
     5041 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     5101 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     5161 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     5221 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     5281 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     5341 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     5401 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     5461 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     5521 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     5581 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggta
     5641 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     5701 gggggactgg aagggctaat tcactcccaa cgaaaataag atctgctttt tgcttgtact
     5761 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     5821 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     5881 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     5941 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     6001 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     6061 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     6121 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccagttccg
     6181 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     6241 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta gacttttgca
     6301 gagacggccc aaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     6361 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     6421 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     6481 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     6541 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6601 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6661 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6721 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6781 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6841 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6901 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6961 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     7021 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     7081 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     7141 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     7201 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     7261 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     7321 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     7381 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     7441 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     7501 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     7561 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     7621 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     7681 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     7741 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     7801 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7861 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7921 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7981 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     8041 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     8101 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     8161 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     8221 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     8281 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     8341 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     8401 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     8461 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     8521 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     8581 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     8641 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     8701 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     8761 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
     8821 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     8881 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     8941 gtgccaagct g
//
