LOCUS       V020913                 8412 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020913
VERSION     V020913
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..3308
                     /label="Abhd6(NM_025341)"
                     /note="Abhd6(NM_025341)"
                     /gene="Abhd6"
     CDS             3309..3332
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     promoter        3382..3593
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3606..3874
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             3901..4500
                     /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    4507..5095
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5169..5402
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5474..5595
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5614..5749
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(5787..5803)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5811..5827
                     /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(5835..5865)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5880..5901
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6189..6777)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6948..7808)
                     /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(7809..7913)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8387..8403
                     /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 atggatctcg atgtggttaa catgtttgtg attgcgggtg
     2341 ggaccctggc cattccaatc ctggcatttg ttgcgtcttt cctcctgtgg ccttcagcac
     2401 tgataagaat ctattattgg tactggcgga ggacactggg catgcaagtt cgctacgcac
     2461 accatgagga ctatcagttc tgttactcct tccggggcag gccaggacac aagccatcca
     2521 tccttatgct ccatggattc tccgcacaca aggacatgtg gctcagcgtg gtcaagttcc
     2581 ttccgaagaa cctgcacttg gtctgtgtgg acatgcctgg gcatgaaggc accacccgct
     2641 cctccctgga tgacctgtcc atagtggggc aagttaaaag gatacatcag tttgtagaat
     2701 gccttaagct gaacaaaaag ccctttcacc ttataggcac ctccatgggt ggccacgtgg
     2761 ctggagtata tgccgcttac tacccatctg atgtctgcag cctgtctctc gtgtgtcctg
     2821 ctggcctgca gtactcaact gacaatcctt ttgtacaacg gctcaaagag ctggaggagt
     2881 cagctgccat tcagaagatt cccttgatcc catccacccc ggaagagatg agtgagatgc
     2941 tgcagctctg ctcctatgtc cgcttcaagg tgccccagca gatccttcaa ggtcttgtcg
     3001 atgttcgcat tcctcataac agcttctacc ggaaattgtt tttggaaatc gtcaatgaga
     3061 aatccagata ctctctgcat gagaatatgg acaagatcaa ggtcccgaca cagatcattt
     3121 gggggaaaca agaccaggtg cttgatgtgt ccggggcaga catattagcc aagtcaatct
     3181 ctaactccca ggtagaggtt ctggaaaact gtggccattc ggtagtgatg gagagaccga
     3241 ggaagacagc caagctcatt gtcgactttt tagcttctgt gcataacaca gacaacaaga
     3301 agctgaacga ttacaaggac gacgatgaca agtaggatcc gcggccgcga aggatctgcg
     3361 atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
     3421 ggggaggggt cggcaattga acgggtgcct agagaaggtg gcgcggggta aactgggaaa
     3481 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
     3541 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca cagctgaagc
     3601 ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc cgccatccac
     3661 gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg
     3721 tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc ccttggagcc
     3781 tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa ctctacgtct
     3841 ttgtttcgtt ttctgttctg cgccgttaca gatccaagct gtgaccggcg cctacgctag
     3901 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta
     3961 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgacccggac
     4021 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     4081 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     4141 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     4201 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
     4261 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
     4321 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
     4381 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     4441 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
     4501 gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     4561 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     4621 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     4681 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     4741 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     4801 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     4861 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     4921 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     4981 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     5041 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctggt
     5101 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     5161 ggggggactg gaagggctaa ttcactccca acgaaaataa gatctgcttt ttgcttgtac
     5221 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     5281 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     5341 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     5401 cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc
     5461 agagagtgag aggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5521 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5581 catcaatgta tcttatcatg tctggctcta gctatcccgc ccctaactcc gcccagttcc
     5641 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     5701 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc
     5761 agagacggcc caaattcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     5821 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     5881 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     5941 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     6001 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     6061 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     6121 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     6181 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     6241 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     6301 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     6361 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     6421 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     6481 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     6541 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     6601 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     6661 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     6721 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6781 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6841 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6901 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6961 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     7021 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     7081 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     7141 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     7201 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     7261 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     7321 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     7381 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     7441 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     7501 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     7561 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     7621 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     7681 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     7741 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     7801 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     7861 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     7921 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     7981 aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
     8041 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     8101 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
     8161 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
     8221 gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa
     8281 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     8341 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     8401 agtgccaagc tg
//
