LOCUS       V046379                 9103 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046379
VERSION     V046379
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..3957
                     /label="USH1C(NM_005709)"
                     /note="USH1C(NM_005709)"
                     /gene="USH1C"
     CDS             3964..4029
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4073..4284
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4297..4565
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4592..5191
                     /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    5198..5786
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5860..6093
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6165..6286
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6305..6440
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6478..6494)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6502..6518
                     /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(6526..6556)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6571..6592
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6880..7468)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7639..8499)
                     /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(8500..8604)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9078..9094
                     /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 catggaccga aaagtggccc gagaattccg gcataaggtg
     2341 gattttctga ttgaaaatga tgcagagaag gactatctct atgatgtgct gcgaatgtac
     2401 caccagacca tggacgtggc cgtgctcgtg ggagacctga agctggtcat caatgaaccc
     2461 agccgtctgc ctctgtttga tgccattcgg ccgctgatcc cactgaagca ccaggtggaa
     2521 tatgatcagc tgaccccccg gcgctccagg aagctgaagg aggtgcgtct ggaccgtctg
     2581 caccccgaag gcctcggcct gagtgtgcgt ggtggcctgg agtttggctg tgggctcttc
     2641 atctcccacc tcatcaaagg cggtcaggca gacagcgtcg ggctccaggt aggggacgag
     2701 atcgtccgga tcaatggata ttccatctcc tcctgtaccc atgaggaggt catcaacctc
     2761 attcgaacca agaaaactgt gtccatcaaa gtgagacaca tcggcctgat ccccgtgaaa
     2821 agctctcctg atgagcccct cacttggcag tatgtggatc agtttgtgtc ggaatctggg
     2881 ggcgtgcgag gcagcctggg ctcccctgga aatcgggaaa acaaggagaa gaaggtcttc
     2941 atcagcctgg taggctcccg aggccttggc tgcagcattt ccagcggccc catccagaag
     3001 cctggcatct ttatcagcca tgtgaaacct ggctccctgt ctgctgaggt gggattggag
     3061 ataggggacc agattgtcga agtcaatggc gtcgacttct ctaacctgga tcacaaggag
     3121 gctgtaaatg tgctgaagag tagccgcagc ctgaccatct ccattgtagc tgcagctggc
     3181 cgggagctgt tcatgacaga ccgggagcgg ctggcagagg cgcggcagcg tgagctgcag
     3241 cggcaggagc ttctcatgca gaagcggctg gcgatggagt ccaacaagat cctccaggag
     3301 cagcaggaga tggagcggca aaggagaaaa gaaattgccc agaaggcagc agaggaaaat
     3361 gagagatacc ggaaggagat ggaacagatt gtagaggagg aagagaagtt taagaagcaa
     3421 tgggaagaag actggggctc aaaggaacag ctactcttgc ctaaaaccat cactgctgag
     3481 gtacacccag taccccttcg caagccaaag tatgatcagg gagtggaacc tgagctcgag
     3541 cccgcagatg acctggatgg aggcacggag gagcagggag agcaggattt ccggaaatat
     3601 gaggaaggct ttgaccccta ctctatgttc accccagagc agatcatggg gaaggatgtc
     3661 cggctcctac gcatcaagaa ggagggatcc ttagacctgg ccctggaagg cggtgtggac
     3721 tcccccattg ggaaggtggt cgtttctgct gtgtatgagc ggggagctgc tgagcggcat
     3781 ggtggcattg tgaaagggga cgagatcatg gcaatcaacg gcaagattgt gacagactac
     3841 accctggctg aggctgaggc tgccctgcag aaggcctgga atcagggcgg ggactggatc
     3901 gaccttgtgg ttgccgtctg ccccccaaag gagtatgacg atgagctgac cttcttcgga
     3961 tccgactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac
     4021 gatgacaagt agcggccgcg aaggatctgc gatcgctccg gtgcccgtca gtgggcagag
     4081 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aacgggtgcc
     4141 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt
     4201 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc
     4261 aacgggtttg ccgccagaac acagctgaag cttcgagggg ctcgcatctc tccttcacgc
     4321 gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc
     4381 gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga
     4441 ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt
     4501 tgcctgaccc tgcttgctca actctacgtc tttgtttcgt tttctgttct gcgccgttac
     4561 agatccaagc tgtgaccggc gcctacgcta gatgaccgag tacaagccca cggtgcgcct
     4621 cgccacccgc gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt tcgccgacta
     4681 ccccgccacg cgccacaccg tcgacccgga ccgccacatc gagcgggtca ccgagctgca
     4741 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     4801 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     4861 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     4921 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     4981 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     5041 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
     5101 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     5161 ctggtgcatg acccgcaagc ccggtgcctg agtcgacaat caacctctgg attacaaaat
     5221 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     5281 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     5341 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     5401 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     5461 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     5521 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     5581 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     5641 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     5701 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     5761 gatctccctt tgggccgcct ccccgcctgg tacctttaag accaatgact tacaaggcag
     5821 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc
     5881 aacgaaaata agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag
     5941 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     6001 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     6061 gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc
     6121 agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag
     6181 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     6241 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct
     6301 agctatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     6361 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     6421 aggaggcttt tttggaggcc tagacttttg cagagacggc ccaaattcgt aatcatggtc
     6481 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     6541 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     6601 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     6661 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     6721 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     6781 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     6841 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     6901 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6961 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     7021 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     7081 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     7141 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     7201 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     7261 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     7321 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     7381 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     7441 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     7501 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     7561 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     7621 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7681 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     7741 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     7801 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     7861 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     7921 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     7981 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     8041 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     8101 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     8161 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     8221 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     8281 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     8341 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     8401 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     8461 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     8521 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     8581 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     8641 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     8701 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     8761 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     8821 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     8881 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     8941 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     9001 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     9061 ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctg
//
