LOCUS       V026085                 9663 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026085
VERSION     V026085
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..4517
                     /label="ACSL5(NM_016234)"
                     /note="ACSL5(NM_016234)"
                     /gene="ACSL5"
     CDS             4524..4589
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4633..4844
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4857..5125
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5152..5751
                     /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    5758..6346
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6420..6653
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6725..6846
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6865..7000
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7038..7054)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7062..7078
                     /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(7086..7116)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7131..7152
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7440..8028)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8199..9059)
                     /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(9060..9164)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9638..9654
                     /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 atggacgctc tgaagccacc ctgtctctgg aggaaccacg
     2341 agcgagggaa gaaggacagg gactcgtgtg gcaggaagaa ctcagagccg ggaagccccc
     2401 attcactaga agcactgaga gatgcggccc cctcgcaggg tctgaatttc ctgctgctgt
     2461 tcacaaagat gctttttatc tttaactttt tgttttcccc acttccgacc ccggcgttga
     2521 tctgcatcct gacatttgga gctgccatct tcttgtggct gatcaccaga cctcaacccg
     2581 tcttacctct tcttgacctg aacaatcagt ctgtgggaat tgagggagga gcacggaagg
     2641 gggtttccca gaagaacaat gacctaacaa gttgctgctt ctcagatgcc aagactatgt
     2701 atgaggtttt ccaaagagga ctcgctgtgt ctgacaatgg gccctgcttg ggatatagaa
     2761 aaccaaacca gccctacaga tggctatctt acaaacaggt gtctgataga gcagagtacc
     2821 tgggttcctg tctcttgcat aaaggttata aatcatcacc agaccagttt gtcggcatct
     2881 ttgctcagaa taggccagag tggatcatct ccgaattggc ttgttacacg tactctatgg
     2941 tagctgtacc tctgtatgac accttgggac cagaagccat cgtacatatt gtcaacaagg
     3001 ctgatatcgc catggtgatc tgtgacacac cccaaaaggc attggtgctg atagggaatg
     3061 tagagaaagg cttcaccccg agcctgaagg tgatcatcct tatggacccc tttgatgatg
     3121 acctgaagca aagaggggag aagagtggaa ttgagatctt atccctatat gatgctgaga
     3181 acctaggcaa agagcacttc agaaaacctg tgcctcctag cccagaagac ctgagcgtca
     3241 tctgcttcac cagtgggacc acaggtgacc ccaaaggagc catgataacc catcaaaata
     3301 ttgtttcaaa tgctgctgcc tttctcaaat gtgtggagca tgcttatgag cccactcctg
     3361 atgatgtggc catatcctac ctccctctgg ctcatatgtt tgagaggatt gtacaggctg
     3421 ttgtgtacag ctgtggagcc agagttggat tcttccaagg ggatattcgg ttgctggctg
     3481 acgacatgaa gactttgaag cccacattgt ttcccgcggt gcctcgactc cttaacagga
     3541 tctacgataa ggtacaaaat gaggccaaga cacccttgaa gaagttcttg ttgaagctgg
     3601 ctgtttccag taaattcaaa gagcttcaaa agggtatcat caggcatgat agtttctggg
     3661 acaagctcat ctttgcaaag atccaggaca gcctgggcgg aagggttcgt gtaattgtca
     3721 ctggagctgc ccccatgtcc acttcagtca tgacattctt ccgggcagca atgggatgtc
     3781 aggtgtatga agcttatggt caaacagaat gcacaggtgg ctgtacattt acattacctg
     3841 gggactggac atcaggtcac gttggggtgc ccctggcttg caattacgtg aagctggaag
     3901 atgtggctga catgaactac tttacagtga ataatgaagg agaggtctgc atcaagggta
     3961 caaacgtgtt caaaggatac ctgaaggacc ctgagaagac acaggaagcc ctggacagtg
     4021 atggctggct tcacacagga gacattggtc gctggctccc gaatggaact ctgaagatca
     4081 tcgaccgtaa aaagaacatt ttcaagctgg cccaaggaga atacattgca ccagagaaga
     4141 tagaaaatat ctacaacagg agtcaaccag tgttacaaat ttttgtacac ggggagagct
     4201 tacggtcatc cttagtagga gtggtggttc ctgacacaga tgtacttccc tcatttgcag
     4261 ccaagcttgg ggtgaagggc tcctttgagg aactgtgcca aaaccaagtt gtaagggaag
     4321 ccattttaga agacttgcag aaaattggga aagaaagtgg ccttaaaact tttgaacagg
     4381 tcaaagccat ttttcttcat ccagagccat tttccattga aaatgggctc ttgacaccaa
     4441 cattgaaagc aaagcgagga gagctttcca aatactttcg gacccaaatt gacagcctgt
     4501 atgagcacat ccaggatgga tccgactaca aagaccatga cggtgattat aaagatcatg
     4561 acatcgacta caaggatgac gatgacaagt agcggccgcg aaggatctgc gatcgctccg
     4621 gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg
     4681 tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg
     4741 tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg
     4801 ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acagctgaag cttcgagggg
     4861 ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg ccgccatcca cgccggttga
     4921 gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa
     4981 gtttaaagct caggtcgaga ccgggccttt gtccggcgct cccttggagc ctacctagac
     5041 tcagccggct ctccacgctt tgcctgaccc tgcttgctca actctacgtc tttgtttcgt
     5101 tttctgttct gcgccgttac agatccaagc tgtgaccggc gcctacgcta gatgaccgag
     5161 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt acgcaccctc
     5221 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgacccgga ccgccacatc
     5281 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     5341 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     5401 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     5461 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     5521 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     5581 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc
     5641 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     5701 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg agtcgacaat
     5761 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     5821 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     5881 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     5941 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     6001 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     6061 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     6121 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     6181 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     6241 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     6301 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg tacctttaag
     6361 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact
     6421 ggaagggcta attcactccc aacgaaaata agatctgctt tttgcttgta ctgggtctct
     6481 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
     6541 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
     6601 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag
     6661 ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga
     6721 gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     6781 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     6841 atcttatcat gtctggctct agctatcccg cccctaactc cgcccagttc cgcccattct
     6901 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     6961 gagctattcc agaagtagtg aggaggcttt tttggaggcc tagacttttg cagagacggc
     7021 ccaaattcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     7081 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     7141 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     7201 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     7261 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     7321 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     7381 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     7441 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     7501 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     7561 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     7621 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     7681 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     7741 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     7801 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     7861 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     7921 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     7981 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     8041 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     8101 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     8161 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     8221 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     8281 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     8341 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     8401 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     8461 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     8521 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     8581 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     8641 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     8701 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     8761 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     8821 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     8881 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     8941 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     9001 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     9061 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     9121 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     9181 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     9241 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     9301 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     9361 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     9421 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     9481 aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     9541 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     9601 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgccaag
     9661 ctg
//
