LOCUS       V045153                 9013 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045153
VERSION     V045153
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..3867
                     /label="PLIN1(NM_002666)"
                     /note="PLIN1(NM_002666)"
                     /gene="PLIN1"
     CDS             3874..3939
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3983..4194
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4207..4475
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4502..5101
                     /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    5108..5696
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5770..6003
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6075..6196
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6215..6350
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6388..6404)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6412..6428
                     /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(6436..6466)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6481..6502
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6790..7378)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7549..8409)
                     /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(8410..8514)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8988..9004
                     /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 catggcagtc aacaaaggcc tcaccttgct ggatggagac
     2341 ctccctgagc aggagaatgt gctgcagcgg gtcctgcagc tgccggtggt gagtggcacc
     2401 tgcgaatgct tccagaagac ctacaccagc actaaggaag cccaccccct ggtggcctct
     2461 gtgtgcaatg cctatgagaa gggcgtgcag agcgccagta gcttggctgc ctggagcatg
     2521 gagccggtgg tccgcaggct gtccacccag ttcacagctg ccaatgagct ggcctgccga
     2581 ggcttggacc acctggagga aaagatcccc gccctccagt acccccctga aaagattgct
     2641 tctgagctga aggacaccat ctccacccgc ctccgcagtg ccagaaacag catcagcgtt
     2701 cccatcgcga gcacttcaga caaggtcctg ggggccgctt tggccgggtg cgagcttgcc
     2761 tggggggtgg ccagagacac tgcggaattt gctgccaaca ctcgagctgg ccgactggct
     2821 tctggagggg ccgacttggc cttgggcagc attgagaagg tggtggagta cctcctccct
     2881 ccagacaagg aagagtcagc ccctgctcct ggacaccagc aagcccagaa gtctcccaag
     2941 gccaagccaa gcctcttgag cagggttggg gctctgacca acaccctctc tcgatacacc
     3001 gtgcagacca tggcccgggc cctggagcag ggccacaccg tggccatgtg gatcccaggc
     3061 gtggtgcccc tgagcagcct ggcccagtgg ggtgcctcag tggccatgca ggcggtgtcc
     3121 cggcggagga gcgaagtgcg ggtaccctgg ctgcacagcc tcgcagccgc ccaggaggag
     3181 gatcatgagg accagacaga cacggaggga gaggacacgg aggaggagga agaattggag
     3241 actgaggaga acaagttcag tgaggtagca gccctgccag gccctcgagg cctcctgggt
     3301 ggtgtggcac ataccctgca gaagaccctc cagaccacca tctcggctgt gacatgggca
     3361 cctgcagctg tgctgggcat ggcagggagg gtgctgcacc tcacaccagc ccctgctgtc
     3421 tcctcaacca aggggagggc catgtcccta tcagatgccc tgaagggcgt tactgacaac
     3481 gtggtggaca cagtggtgca ttacgtgccg ctccccaggc tgtcgctgat ggagcccgag
     3541 agcgaattcc gggacatcga caacccacca gccgaggtcg agcgccggga ggcggagcgc
     3601 agagcgtctg gggcgccgtc cgccggcccg gagcccgccc cgcgtctcgc acagccccgc
     3661 cgcagcctgc gcagcgcgca gagccccggc gcgccccccg gcccgggcct ggaggacgaa
     3721 gtcgccacgc ccgcagcgcc gcgcccgggc ttcccggccg tgccccgcga gaagccaaag
     3781 cgcagggtca gcgacagctt cttccggccc agcgtcatgg agcccatcct gggccgcacg
     3841 cattacagcc agctgcgcaa gaagagcgga tccgactaca aagaccatga cggtgattat
     3901 aaagatcatg acatcgacta caaggatgac gatgacaagt agcggccgcg aaggatctgc
     3961 gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg
     4021 gggggagggg tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt aaactgggaa
     4081 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt
     4141 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acagctgaag
     4201 cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg ccgccatcca
     4261 cgccggttga gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc
     4321 gtctaggtaa gtttaaagct caggtcgaga ccgggccttt gtccggcgct cccttggagc
     4381 ctacctagac tcagccggct ctccacgctt tgcctgaccc tgcttgctca actctacgtc
     4441 tttgtttcgt tttctgttct gcgccgttac agatccaagc tgtgaccggc gcctacgcta
     4501 gatgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt
     4561 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgacccgga
     4621 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
     4681 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
     4741 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
     4801 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
     4861 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
     4921 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
     4981 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
     5041 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg
     5101 agtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     5161 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     5221 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     5281 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     5341 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     5401 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     5461 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     5521 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     5581 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     5641 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
     5701 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
     5761 aggggggact ggaagggcta attcactccc aacgaaaata agatctgctt tttgcttgta
     5821 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     5881 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     5941 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     6001 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat
     6061 cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     6121 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     6181 tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc cgcccagttc
     6241 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     6301 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc tagacttttg
     6361 cagagacggc ccaaattcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     6421 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     6481 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     6541 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6601 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6661 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6721 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6781 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6841 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6901 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6961 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     7021 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     7081 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     7141 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     7201 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     7261 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     7321 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     7381 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     7441 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     7501 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     7561 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7621 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7681 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7741 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7801 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7861 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7921 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7981 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     8041 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     8101 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     8161 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     8221 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     8281 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     8341 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     8401 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     8461 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     8521 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     8581 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     8641 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     8701 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     8761 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     8821 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
     8881 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     8941 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     9001 cagtgccaag ctg
//
