LOCUS       V026084                 9576 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026084
VERSION     V026084
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..4430
                     /label="LTF(NM_002343)"
                     /note="LTF(NM_002343)"
                     /gene="LTF"
     CDS             4437..4502
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4546..4757
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4770..5038
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5065..5664
                     /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    5671..6259
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6333..6566
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6638..6759
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6778..6913
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6951..6967)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6975..6991
                     /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(6999..7029)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7044..7065
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7353..7941)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8112..8972)
                     /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(8973..9077)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9551..9567
                     /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 atgaaacttg tcttcctcgt cctgctgttc ctcggggccc
     2341 tcggactgtg tctggctggc cgtaggagga gtgttcagtg gtgcgccgta tcccaacccg
     2401 aggccacaaa atgcttccaa tggcaaagga atatgagaaa agtgcgtggc cctcctgtca
     2461 gctgcataaa gagagactcc cccatccagt gtatccaggc cattgcggaa aacagggccg
     2521 atgctgtgac ccttgatggt ggtttcatat acgaggcagg cctggccccc tacaaactgc
     2581 gacctgtagc ggcggaagtc tacgggaccg aaagacagcc acgaactcac tattatgccg
     2641 tggctgtggt gaagaagggc ggcagctttc agctgaacga actgcaaggt ctgaagtcct
     2701 gccacacagg ccttcgcagg accgctggat ggaatgtccc tatagggaca cttcgtccat
     2761 tcttgaattg gacgggtcca cctgagccca ttgaggcagc tgtggccagg ttcttctcag
     2821 ccagctgtgt tcccggtgca gataaaggac agttccccaa cctgtgtcgc ctgtgtgcgg
     2881 ggacagggga aaacaaatgt gccttctcct cccaggaacc gtacttcagc tactctggtg
     2941 ccttcaagtg tctgagagac ggggctggag acgtggcttt tatcagagag agcacagtgt
     3001 ttgaggacct gtcagacgag gctgaaaggg acgagtatga gttactctgc ccagacaaca
     3061 ctcggaagcc agtggacaag ttcaaagact gccatctggc ccgggtccct tctcatgccg
     3121 ttgtggcacg aagtgtgaat ggcaaggagg atgccatctg gaatcttctc cgccaggcac
     3181 aggaaaagtt tggaaaggac aagtcaccga aattccagct ctttggctcc cctagtgggc
     3241 agaaagatct gctgttcaag gactctgcca ttgggttttc gagggtgccc ccgaggatag
     3301 attctgggct gtaccttggc tccggctact tcactgccat ccagaacttg aggaaaagtg
     3361 aggaggaagt ggctgcccgg cgtgcgcggg tcgtgtggtg tgcggtgggc gagcaggagc
     3421 tgcgcaagtg taaccagtgg agtggcttga gcgaaggcag cgtgacctgc tcctcggcct
     3481 ccaccacaga ggactgcatc gccctggtgc tgaaaggaga agctgatgcc atgagtttgg
     3541 atggaggata tgtgtacact gcaggcaaat gtggtttggt gcctgtcctg gcagagaact
     3601 acaaatccca acaaagcagt gaccctgatc ctaactgtgt ggatagacct gtggaaggat
     3661 atcttgctgt ggcggtggtt aggagatcag acactagcct tacctggaac tctgtgaaag
     3721 gcaagaagtc ctgccacacc gccgtggaca ggactgcagg ctggaatatc cccatgggcc
     3781 tgctcttcaa ccagacgggc tcctgcaaat ttgatgaata tttcagtcaa agctgtgccc
     3841 ctgggtctga cccgagatct aatctctgtg ctctgtgtat tggcgacgag cagggtgaga
     3901 ataagtgcgt gcccaacagc aacgagagat actacggcta cactggggct ttccggtgcc
     3961 tggctgagaa tgctggagac gttgcatttg tgaaagatgt cactgtcttg cagaacactg
     4021 atggaaataa caatgaggca tgggctaagg atttgaagct ggcagacttt gcgctgctgt
     4081 gcctcgatgg caaacggaag cctgtgactg aggctagaag ctgccatctt gccatggccc
     4141 cgaatcatgc cgtggtgtct cggatggata aggtggaacg cctgaaacag gtgttgctcc
     4201 accaacaggc taaatttggg agaaatggat ctgactgccc ggacaagttt tgcttattcc
     4261 agtctgaaac caaaaacctt ctgttcaatg acaacactga gtgtctggcc agactccatg
     4321 gcaaaacaac atatgaaaaa tatttgggac cacagtatgt cgcaggcatt actaatctga
     4381 aaaagtgctc aacctccccc ctcctggaag cctgtgaatt cctcaggaag ggatccgact
     4441 acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca
     4501 agtagcggcc gcgaaggatc tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat
     4561 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaacgggt gcctagagaa
     4621 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
     4681 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
     4741 ttgccgccag aacacagctg aagcttcgag gggctcgcat ctctccttca cgcgcccgcc
     4801 gccctacctg aggccgccat ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg
     4861 gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg agaccgggcc
     4921 tttgtccggc gctcccttgg agcctaccta gactcagccg gctctccacg ctttgcctga
     4981 ccctgcttgc tcaactctac gtctttgttt cgttttctgt tctgcgccgt tacagatcca
     5041 agctgtgacc ggcgcctacg ctagatgacc gagtacaagc ccacggtgcg cctcgccacc
     5101 cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
     5161 acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
     5221 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
     5281 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
     5341 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc
     5401 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc
     5461 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag
     5521 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag
     5581 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
     5641 atgacccgca agcccggtgc ctgagtcgac aatcaacctc tggattacaa aatttgtgaa
     5701 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     5761 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     5821 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     5881 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     5941 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     6001 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     6061 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
     6121 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     6181 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     6241 ctttgggccg cctccccgcc tggtaccttt aagaccaatg acttacaagg cagctgtaga
     6301 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaaa
     6361 ataagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga
     6421 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     6481 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     6541 ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt
     6601 ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa
     6661 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     6721 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc
     6781 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     6841 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     6901 ttttttggag gcctagactt ttgcagagac ggcccaaatt cgtaatcatg gtcatagctg
     6961 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     7021 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     7081 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     7141 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     7201 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     7261 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     7321 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     7381 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     7441 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     7501 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     7561 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     7621 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     7681 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     7741 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     7801 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     7861 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     7921 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     7981 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     8041 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     8101 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     8161 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     8221 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     8281 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     8341 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     8401 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     8461 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     8521 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     8581 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     8641 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     8701 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     8761 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     8821 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     8881 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     8941 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     9001 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     9061 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     9121 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     9181 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     9241 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     9301 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     9361 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca
     9421 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     9481 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     9541 tcacgacgtt gtaaaacgac ggccagtgcc aagctg
//
