LOCUS       V022408                 9516 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022408
VERSION     V022408
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             2301..4370
                     /label="CAPN9(NM_006615)"
                     /note="CAPN9(NM_006615)"
                     /gene="CAPN9"
     CDS             4377..4442
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4486..4697
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4710..4978
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5005..5604
                     /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    5611..6199
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6273..6506
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6578..6699
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6718..6853
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6891..6907)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6915..6931
                     /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(6939..6969)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6984..7005
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7293..7881)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8052..8912)
                     /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(8913..9017)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9491..9507
                     /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 tgaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagattcta
     2281 gagctagcga attcgccacc atgccttacc tctaccgggc cccagggcct caggcacacc
     2341 cggttcccaa ggacgcccgg atcacccact cctcaggcca gagctttgag caaatgaggc
     2401 aggagtgcct gcagagaggc accctgtttg aggatgcaga cttcccagcc agcaattcct
     2461 ccctgttcta cagtgagagg ccgcagatcc cctttgtgtg gaaacgacca ggggaaatcg
     2521 tgaaaaaccc agaattcatt cttggagggg ccaccaggac tgatatctgc cagggagagc
     2581 tgggagactg ctggctatta gccgccatcg cctcccttac gcttaatcaa aaagcactgg
     2641 ccagagtcat cccccaggac caaagctttg gccctggtta tgccgggata ttccatttcc
     2701 agttctggca gcacagtgag tggctggacg tggtgatcga tgaccgcctg cccaccttca
     2761 gggaccgctt ggttttcctc cactctgccg accacaacga gttctggagc gccttgctgg
     2821 aaaaagccta cgccaagcta aatgggagct atgaagctct gaagggaggc agcgccatcg
     2881 aggccatgga agacttcact gggggtgtgg cagagacctt ccaaactaaa gaggcccccg
     2941 agaacttcta tgagattcta gagaaggctt tgaagagagg ctccctgctg ggctgcttca
     3001 ttgataccag aagtgctgca gaatctgagg cccggacgcc gtttggtctt attaagggtc
     3061 atgcctacag tgtaacggga attgaccagg taagcttccg aggccagaga atcgagctca
     3121 tccgaatccg gaacccttgg ggccaggttg agtggaacgg gtcgtggagc gacagttctc
     3181 cggagtggcg ttctgttggt ccagctgagc agaagcgtct gtgtcacact gctctggatg
     3241 atggggaatt ctggatggca tttaaggact tcaaggccca ctttgataaa gtggagatct
     3301 gcaacctcac tcccgatgcc ctggaggaag acgcgatcca caaatgggag gtgacggtcc
     3361 atcagggaag ctgggttcgc ggctccacgg ctgggggctg ccgcaatttc ctggatacct
     3421 tttggaccaa tccacaaata aaattgtctc tgactgagaa agatgagggg caggaggagt
     3481 gtagtttcct tgtagccctg atgcagaaag atagaaggaa actcaagaga tttggtgcca
     3541 atgtgctgac aatcggctat gccatttatg agtgccctga caaagacgaa cacctgaaca
     3601 aagacttctt cagataccac gcttctcggg ccagaagcaa gacgttcatc aacctgagag
     3661 aagtctccga ccggttcaag ctgccccctg gggagtacat cctgattccc agcacttttg
     3721 agccccacca ggaagctgat ttctgtctga gaatcttttc agagaaaaaa gccattaccc
     3781 gggatatgga tggaaatgta gacattgacc ttcctgagcc tccaaagcca actccacctg
     3841 accaggagac agaggaggag cagcggtttc gggctctgtt tgaacaagtc gctggtgagg
     3901 acatggaggt gacagcagag gaacttgagt atgttttaaa tgctgtgctg caaaagaaaa
     3961 aggacatcaa attcaagaag ctaagcctga tctcctgtaa aaacatcatt tccctgatgg
     4021 acaccagcgg caatgggaag ctggagtttg atgaattcaa agtgttctgg gacaagctga
     4081 agcagtggat taaccttttc cttcggtttg atgctgacaa gtccggcacc atgtctacct
     4141 atgaactacg gactgcactg aaagctgcag gctttcagct gagcagccac ctcctgcagc
     4201 tgattgtgct caggtatgcg gatgaggagc tccagctgga cttcgatgac ttcctcaact
     4261 gcctggtccg gctggagaat gcgagccggg tgttccaggc tctcagtaca aagaacaagg
     4321 agttcattca tctcaatata aatgagttca tccatttgac aatgaacatc ggatccgact
     4381 acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca
     4441 agtagcggcc gcgaaggatc tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat
     4501 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaacgggt gcctagagaa
     4561 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
     4621 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
     4681 ttgccgccag aacacagctg aagcttcgag gggctcgcat ctctccttca cgcgcccgcc
     4741 gccctacctg aggccgccat ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg
     4801 gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg agaccgggcc
     4861 tttgtccggc gctcccttgg agcctaccta gactcagccg gctctccacg ctttgcctga
     4921 ccctgcttgc tcaactctac gtctttgttt cgttttctgt tctgcgccgt tacagatcca
     4981 agctgtgacc ggcgcctacg ctagatgacc gagtacaagc ccacggtgcg cctcgccacc
     5041 cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
     5101 acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
     5161 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
     5221 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
     5281 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc
     5341 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc
     5401 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag
     5461 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag
     5521 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
     5581 atgacccgca agcccggtgc ctgagtcgac aatcaacctc tggattacaa aatttgtgaa
     5641 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     5701 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     5761 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     5821 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     5881 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     5941 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     6001 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
     6061 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     6121 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     6181 ctttgggccg cctccccgcc tggtaccttt aagaccaatg acttacaagg cagctgtaga
     6241 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaaa
     6301 ataagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga
     6361 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     6421 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     6481 ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt
     6541 ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa
     6601 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     6661 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc
     6721 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     6781 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     6841 ttttttggag gcctagactt ttgcagagac ggcccaaatt cgtaatcatg gtcatagctg
     6901 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     6961 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     7021 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     7081 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     7141 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     7201 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     7261 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     7321 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     7381 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     7441 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     7501 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     7561 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     7621 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     7681 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     7741 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     7801 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     7861 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     7921 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     7981 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     8041 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     8101 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     8161 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     8221 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     8281 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     8341 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     8401 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     8461 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     8521 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     8581 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     8641 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     8701 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     8761 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     8821 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     8881 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     8941 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     9001 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     9061 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     9121 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     9181 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     9241 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     9301 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca
     9361 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     9421 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     9481 tcacgacgtt gtaaaacgac ggccagtgcc aagctg
//
