LOCUS       V046369                 9004 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046369
VERSION     V046369
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..3858
                     /label="LAP3(NM_015907)"
                     /note="LAP3(NM_015907)"
                     /gene="LAP3"
     CDS             3865..3930
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3974..4185
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4198..4466
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4493..5092
                     /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    5099..5687
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5761..5994
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6066..6187
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6206..6341
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6379..6395)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6403..6419
                     /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(6427..6457)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6472..6493
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6781..7369)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7540..8400)
                     /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(8401..8505)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8979..8995
                     /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 catgttcttg ctgcctcttc cggctgcggg gcgagtagtc
     2341 gtccgacgtc tggccgtgag acgtttcggg agccggagtc tctccaccgc agacatgacg
     2401 aagggccttg ttttaggaat ctattccaaa gaaaaagaag atgatgtgcc acagttcaca
     2461 agtgcaggag agaattttga taaattgtta gctggaaagc tgagagagac tttgaacata
     2521 tctggaccac ctctgaaggc agggaagact cgaacctttt atggtctgca tcaggacttc
     2581 cccagcgtgg tgctagttgg cctcggcaaa aaggcagctg gaatcgacga acaggaaaac
     2641 tggcatgaag gcaaagaaaa catcagagct gctgttgcag cggggtgcag gcagattcaa
     2701 gacctggagc tctcgtctgt ggaggtggat ccctgtggag acgctcaggc tgctgcggag
     2761 ggagcggtgc ttggtctcta tgaatacgat gacctaaagc aaaaaaagaa gatggctgtg
     2821 tcggcaaagc tctatggaag tggggatcag gaggcctggc agaaaggagt cctgtttgct
     2881 tctgggcaga acttggcacg ccaattgatg gagacgccag ccaatgagat gacgccaacc
     2941 agatttgctg aaattattga gaagaatctc aaaagtgcta gtagtaaaac cgaggtccat
     3001 atcagaccca agtcttggat tgaggaacag gcaatgggat cattcctcag tgtggccaaa
     3061 ggatctgacg agcccccagt cttcttggaa attcactaca aaggcagccc caatgcaaac
     3121 gaaccacccc tggtgtttgt tgggaaagga attacctttg acagtggtgg tatctccatc
     3181 aaggcttctg caaatatgga cctcatgagg gctgacatgg gaggagctgc aactatatgc
     3241 tcagccatcg tgtctgctgc aaagcttaat ttgcccatta atattatagg tctggcccct
     3301 ctttgtgaaa atatgcccag cggcaaggcc aacaagccgg gggatgttgt tagagccaaa
     3361 aacgggaaga ccatccaggt tgataacact gatgctgagg ggaggctcat actggctgat
     3421 gcgctctgtt acgcacacac gtttaacccg aaggtcatcc tcaatgccgc caccttaaca
     3481 ggtgccatgg atgtagcttt gggatcaggt gccactgggg tctttaccaa ttcatcctgg
     3541 ctctggaaca aactcttcga ggccagcatt gaaacagggg accgtgtctg gaggatgcct
     3601 ctcttcgaac attatacaag acaggttgta gattgccagc ttgctgatgt taacaacatt
     3661 ggaaaataca gatctgcagg agcatgtaca gctgcagcat tcctgaaaga attcgtaact
     3721 catcctaagt gggcacattt agacatagca ggcgtgatga ccaacaaaga tgaagttccc
     3781 tatctacgga aaggcatgac tgggaggccc acaaggactc tcattgagtt cttacttcgt
     3841 ttcagtcaag acaatgctgg atccgactac aaagaccatg acggtgatta taaagatcat
     3901 gacatcgact acaaggatga cgatgacaag tagcggccgc gaaggatctg cgatcgctcc
     3961 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
     4021 gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
     4081 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
     4141 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa gcttcgaggg
     4201 gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc acgccggttg
     4261 agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta
     4321 agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag cctacctaga
     4381 ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg
     4441 ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct agatgaccga
     4501 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg tacgcaccct
     4561 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg accgccacat
     4621 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     4681 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     4741 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     4801 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     4861 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     4921 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     4981 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     5041 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gagtcgacaa
     5101 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     5161 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     5221 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     5281 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     5341 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     5401 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     5461 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     5521 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     5581 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     5641 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa
     5701 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
     5761 tggaagggct aattcactcc caacgaaaat aagatctgct ttttgcttgt actgggtctc
     5821 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     5881 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     5941 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta
     6001 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg
     6061 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     6121 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     6181 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccagtt ccgcccattc
     6241 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     6301 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagactttt gcagagacgg
     6361 cccaaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     6421 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     6481 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     6541 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     6601 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     6661 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     6721 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     6781 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6841 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6901 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6961 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     7021 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     7081 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     7141 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     7201 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     7261 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     7321 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     7381 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     7441 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     7501 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     7561 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     7621 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     7681 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     7741 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     7801 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     7861 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     7921 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     7981 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     8041 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     8101 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     8161 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     8221 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     8281 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     8341 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     8401 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     8461 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     8521 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     8581 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     8641 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     8701 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     8761 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     8821 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     8881 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     8941 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa
     9001 gctg
//
