LOCUS       V025087                 9432 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025087
VERSION     V025087
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..4286
                     /label="ALOX15(NM_001140)"
                     /note="ALOX15(NM_001140)"
                     /gene="ALOX15"
     CDS             4293..4358
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4402..4613
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4626..4894
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4921..5520
                     /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    5527..6115
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6189..6422
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6494..6615
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6634..6769
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6807..6823)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6831..6847
                     /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(6855..6885)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6900..6921
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7209..7797)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7968..8828)
                     /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(8829..8933)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9407..9423
                     /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 atgggtctct accgcatccg cgtgtccact ggggcctcgc
     2341 tctatgccgg ttccaacaac caggtgcagc tgtggctggt cggccagcac ggggaggcgg
     2401 cgctcgggaa gcgactgtgg cccgcacggg gcaaggagac agaactcaag gtggaagtac
     2461 cggagtatct ggggccgctg ctgtttgtga aactgcgcaa acggcacctc cttaaggacg
     2521 acgcctggtt ctgcaactgg atctctgtgc agggccccgg agccggggac gaggtcaggt
     2581 tcccttgtta ccgctgggtg gagggcaacg gcgtcctgag cctgcctgaa ggcaccggcc
     2641 gcactgtggg cgaggaccct cagggcctgt tccagaaaca ccgggaagaa gagctggaag
     2701 agagaaggaa gttgtaccgg tggggaaact ggaaggacgg gttaattctg aatatggctg
     2761 gggccaaact atatgacctc cctgtggatg agcgatttct ggaagacaag agagttgact
     2821 ttgaggtttc gctggccaag gggctggccg acctcgctat caaagactct ctaaatgttc
     2881 tgacttgctg gaaggatcta gatgacttca accggatttt ctggtgtggt cagagcaagc
     2941 tggctgagcg cgtgcgggac tcctggaagg aagatgcctt atttgggtac cagtttctta
     3001 atggcgccaa ccccgtggtg ctgaggcgct ctgctcacct tcctgctcgc ctagtgttcc
     3061 ctccaggcat ggaggaactg caggcccagc tggagaagga gctggaggga ggcacactgt
     3121 tcgaagctga cttctccctg ctggatggga tcaaggccaa cgtcattctc tgtagccagc
     3181 agcacctggc tgcccctcta gtcatgctga aattgcagcc tgatgggaaa ctcttgccca
     3241 tggtcatcca gctccagctg ccccgcacag gatccccacc acctcccctt ttcttgccta
     3301 cggatccccc aatggcctgg cttctggcca aatgctgggt gcgcagctct gacttccagc
     3361 tccatgagct gcagtctcat cttctgaggg gacacttgat ggctgaggtc attgttgtgg
     3421 ccaccatgag gtgcctgccg tcgatacatc ctatcttcaa gcttataatt ccccacctgc
     3481 gatacaccct ggaaattaac gtccgggcca ggactgggct ggtctctgac atgggaattt
     3541 tcgaccagat aatgagcact ggtgggggag gccacgtgca gctgctcaag caagctggag
     3601 ccttcctaac ctacagctcc ttctgtcccc ctgatgactt ggccgaccgg gggctcctgg
     3661 gagtgaagtc ttccttctat gcccaagatg cgctgcggct ctgggaaatc atctatcggt
     3721 atgtggaagg aatcgtgagt ctccactata agacagacgt ggctgtgaaa gacgacccag
     3781 agctgcagac ctggtgtcga gagatcactg aaatcgggct gcaaggggcc caggaccgag
     3841 ggtttcctgt ctctttacag gctcgggacc aggtttgcca ctttgtcacc atgtgtatct
     3901 tcacctgcac cggccaacac gcctctgtgc acctgggcca gctggactgg tactcttggg
     3961 tgcctaatgc accctgcacg atgcggctgc ccccgccaac caccaaggat gcaacgctgg
     4021 agacagtgat ggcgacactg cccaacttcc accaggcttc tctccagatg tccatcactt
     4081 ggcagctggg cagacgccag cccgttatgg tggctgtggg ccagcatgag gaggagtatt
     4141 tttcgggccc tgagcctaag gctgtgctga agaagttcag ggaggagctg gctgccctgg
     4201 ataaggaaat tgagatccgg aatgcaaagc tggacatgcc ctacgagtac ctgcggccca
     4261 gcgtggtgga aaacagtgtg gccatcggat ccgactacaa agaccatgac ggtgattata
     4321 aagatcatga catcgactac aaggatgacg atgacaagta gcggccgcga aggatctgcg
     4381 atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
     4441 ggggaggggt cggcaattga acgggtgcct agagaaggtg gcgcggggta aactgggaaa
     4501 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
     4561 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca cagctgaagc
     4621 ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc cgccatccac
     4681 gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg
     4741 tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc ccttggagcc
     4801 tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa ctctacgtct
     4861 ttgtttcgtt ttctgttctg cgccgttaca gatccaagct gtgaccggcg cctacgctag
     4921 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta
     4981 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgacccggac
     5041 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     5101 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     5161 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     5221 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
     5281 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
     5341 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
     5401 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     5461 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
     5521 gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     5581 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     5641 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     5701 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     5761 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     5821 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     5881 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     5941 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     6001 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     6061 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctggt
     6121 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     6181 ggggggactg gaagggctaa ttcactccca acgaaaataa gatctgcttt ttgcttgtac
     6241 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     6301 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     6361 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     6421 cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc
     6481 agagagtgag aggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     6541 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     6601 catcaatgta tcttatcatg tctggctcta gctatcccgc ccctaactcc gcccagttcc
     6661 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     6721 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc
     6781 agagacggcc caaattcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     6841 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     6901 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     6961 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     7021 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     7081 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     7141 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     7201 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     7261 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     7321 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     7381 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     7441 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     7501 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     7561 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     7621 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     7681 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     7741 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     7801 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     7861 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     7921 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     7981 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     8041 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     8101 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     8161 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     8221 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     8281 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     8341 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     8401 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     8461 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     8521 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     8581 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     8641 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     8701 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     8761 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     8821 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     8881 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     8941 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     9001 aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
     9061 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     9121 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
     9181 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
     9241 gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa
     9301 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     9361 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     9421 agtgccaagc tg
//
