LOCUS       V036809                 9286 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036809
VERSION     V036809
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..4140
                     /label="NPLOC4(NM_001369698)"
                     /note="NPLOC4(NM_001369698)"
                     /gene="NPLOC4"
     CDS             4147..4212
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4256..4467
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4480..4748
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4775..5374
                     /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    5381..5969
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6043..6276
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6348..6469
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6488..6623
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6661..6677)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6685..6701
                     /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(6709..6739)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6754..6775
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7063..7651)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7822..8682)
                     /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(8683..8787)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9261..9277
                     /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 catggccgag agcatcataa ttcgtgtcca gtccccggat
     2341 ggagtgaagc ggatcacagc aacaaagaga gaaacagcag caacattttt gaaaaaggtt
     2401 gcaaaggagt ttggcttcca aaataatggc ttctcggttt acatcaatag aaacaagacc
     2461 ggagagataa cagcctcctc caacaaatcc ctcaacttgc taaaaatcaa gcatggcgat
     2521 ttgttgttcc tgtttccctc gagccttgct gggccctcat ctgaaatgga gacgtcagtt
     2581 ccaccgggct tcaaagtctt tggcgctccc aacgtggtgg aggatgagat tgatcagtac
     2641 ctcagcaaac aggacgggaa gatttacaga agccgagacc cacagctatg ccgccacggc
     2701 cctttgggga aatgcgtgca ctgcgtccct ctagagccat tcgatgagga ctatctaaac
     2761 catctcgagc ctcccgtgaa gcacatgtcc ttccacgcct acatccggaa gctgactgga
     2821 ggggctgaca aggggaagtt tgttgccctg gagaacatca gctgcaagat taagtcaggg
     2881 tgcgaggggc acctcccgtg gccgaatggc atctgtacta agtgccagcc gagcgccatc
     2941 acgctgaaca gacagaagta caggcatgtg gacaatatca tgtttgagaa tcacaccgtc
     3001 gctgaccgct ttcttgactt ctggagaaag acagggaacc agcattttgg gtacttatac
     3061 ggacggtaca cggagcacaa agacattccc cttggcatca gggctgaagt ggctgcgatt
     3121 tatgagccac ctcagattgg tacacagaac agcttggagc ttcttgagga tccaaaagct
     3181 gaagtggtcg atgaaattgc tgccaaactt ggcctgcgga aggttggctg gatatttaca
     3241 gacctcgtct cagaagatac ccgaaagggt accgtccgct acagtcgaaa taaggacacc
     3301 tatttcctaa gttcagaaga gtgcatcact gcaggagact tccagaacaa gcatcccaac
     3361 atgtgccggc tctctccaga cggacatttt ggatccaagt ttgttactgc agtggctaca
     3421 ggtctctctc ttgcaggtgg tcctgacaac caagtccact ttgaagggta ccaggtgtcc
     3481 aatcagtgta tggcactggt ccgtgatgag tgtttgctgc catgcaagga cgccccggag
     3541 cttggctacg ccaaggagtc tagcagtgag cagtacgtgc ctgatgtgtt ttataaggac
     3601 gtagacaagt ttggcaacga gatcacccag ctggcccggc ccctgcctgt ggagtatctc
     3661 atcatagaca tcacaacaac tttccccaag gatccagttt acactttttc tatttcgcaa
     3721 aatccatttc ctattgaaaa ccgggatgta ttgggtgaga cacaggactt ccatagcttg
     3781 gccacctatt tgtctcagaa tacctcatct gtgttcttgg ataccatctc agatttccac
     3841 ctcttgctgt tcctggtcac caatgaagtt atgcctctgc aggacagcat cagcttgctg
     3901 ctggaggccg tgcggaccag aaatgaggag ctcgcccaga catggaagag gtctgagcag
     3961 tgggccacca tcgagcagct gtgcagcaca gttggcgggc agctcccagg tctccatgag
     4021 tacggcgccg tcgggggctc cacacacacg gccactgcag ccatgtgggc ctgtcagcac
     4081 tgcacgttca tgaaccagcc aggcacaggc cactgcgaga tgtgcagcct ccccaggacc
     4141 ggatccgact acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat
     4201 gacgatgaca agtagcggcc gcgaaggatc tgcgatcgct ccggtgcccg tcagtgggca
     4261 gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaacgggt
     4321 gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt
     4381 tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt
     4441 cgcaacgggt ttgccgccag aacacagctg aagcttcgag gggctcgcat ctctccttca
     4501 cgcgcccgcc gccctacctg aggccgccat ccacgccggt tgagtcgcgt tctgccgcct
     4561 cccgcctgtg gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg
     4621 agaccgggcc tttgtccggc gctcccttgg agcctaccta gactcagccg gctctccacg
     4681 ctttgcctga ccctgcttgc tcaactctac gtctttgttt cgttttctgt tctgcgccgt
     4741 tacagatcca agctgtgacc ggcgcctacg ctagatgacc gagtacaagc ccacggtgcg
     4801 cctcgccacc cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg cgttcgccga
     4861 ctaccccgcc acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg tcaccgagct
     4921 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     4981 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     5041 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     5101 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     5161 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     5221 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     5281 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     5341 cacctggtgc atgacccgca agcccggtgc ctgagtcgac aatcaacctc tggattacaa
     5401 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     5461 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     5521 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     5581 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     5641 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     5701 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     5761 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
     5821 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     5881 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     5941 tcggatctcc ctttgggccg cctccccgcc tggtaccttt aagaccaatg acttacaagg
     6001 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     6061 cccaacgaaa ataagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
     6121 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     6181 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     6241 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta
     6301 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg
     6361 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     6421 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc
     6481 tctagctatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     6541 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     6601 gtgaggaggc ttttttggag gcctagactt ttgcagagac ggcccaaatt cgtaatcatg
     6661 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     6721 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     6781 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     6841 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6901 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6961 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     7021 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     7081 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     7141 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     7201 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     7261 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     7321 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7381 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     7441 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7501 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7561 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     7621 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     7681 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     7741 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     7801 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     7861 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     7921 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     7981 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     8041 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     8101 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     8161 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     8221 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     8281 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     8341 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     8401 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     8461 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     8521 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     8581 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     8641 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     8701 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     8761 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     8821 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     8881 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     8941 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     9001 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     9061 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     9121 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     9181 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     9241 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctg
//
