LOCUS       V036882                 9772 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036882
VERSION     V036882
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..4626
                     /label="Tnfaip3(NM_009397)"
                     /note="Tnfaip3(NM_009397)"
                     /gene="Tnfaip3"
     CDS             4633..4698
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4742..4953
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4966..5234
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5261..5860
                     /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    5867..6455
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6529..6762
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6834..6955
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6974..7109
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7147..7163)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7171..7187
                     /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(7195..7225)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7240..7261
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7549..8137)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8308..9168)
                     /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(9169..9273)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9747..9763
                     /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 catggctgaa caacttcttc ctcaggcttt gtatttgagc
     2341 aatatgcgga aagctgtgaa gatacgagag agaaccccag aagacatttt caaacctacc
     2401 aatgggatca tctatcactt taaaaccatg caccgataca cgctggagat gttcagaaca
     2461 tgccagtttt gcccacagtt ccgagagatc atccacaaag cacttattga cagaagtgtc
     2521 caggcttccc tggaaagcca gaagaagctc aactggtgtc gtgaagtcag gaagctcgtg
     2581 gctctgaaaa ccaatggtga tggaaactgc ctcatgcatg cagcttgtca gtacatgtgg
     2641 ggtgttcagg atactgacct ggtcctgagg aaggccctct gcagcaccct taaggagaca
     2701 gacactcgga actttaaatt ccgctggcag ctggaatctc tgaaatctca ggaatttgtg
     2761 gaaacaggac tttgctacga cactcggaac tggaatgacg aatgggacaa cttggtcaaa
     2821 atggcatcag cagacacacc tgcagcccga agtggacttc agtacaattc cctggaagaa
     2881 atccacatat ttgtcctcag caacatcctc agaagaccca tcattgtcat ttcagacaaa
     2941 atgctaagaa gtttggaatc tggttccaat tttgctcctt tgaaagtggg tgggatttat
     3001 ctgcctcttc actggcctgc ccaggagtgt tacagatatc ccatcgtcct aggctatgac
     3061 agccagcact ttgtacccct ggtgaccctg aaggacagtg gacctgaact tcgcgctgtt
     3121 ccacttgtta acagagaccg gggtaggttt gaagacttaa aagttcactt cttgacagat
     3181 cctgagaatg agatgaagga aaagcttcta aaggagtact tgatagtgat ggagatccct
     3241 gtgcaaggct gggaccacgg cacgactcac ctgatcaacg ctgcaaaatt ggatgaagct
     3301 aacttaccca aagaaataaa tttggtagac gattactttg agcttgttca gcacgaatac
     3361 aagaaatggc aggagaacag cgatcaggcc aggagagcgg cacatgcgca gaaccccttg
     3421 gagccttcca caccccagct atcactcatg gatataaaat gtgagacacc caactgtcct
     3481 ttcttcatgt ccgtgaacac tcagccttta tgccacgaat gctcagagag gcgccaaaag
     3541 aatcagagca agctcccaaa gctgaactcg aagctaggcc ctgaaggact cccaggcgtg
     3601 ggacttggct cctcaaactg gagccccgag gaaaccgctg gaggacctca ttcagcccca
     3661 cccacagcac ccagcctttt tctcttcagt gagaccactg caatgaagtg caggagtcct
     3721 gggtgccctt ttactttgaa tgtgcagcat aatggattct gtgagcgttg ccacgcccgg
     3781 cagattaatg ccagccacac cgcagaccct ggaaagtgcc aagcctgcct tcaggatgtc
     3841 actcggacct ttaatggcat ctgcagtacc tgtttcaaaa ggactacagc agagcccagc
     3901 tccagcctca cttccagtat ccctgcctcc tgtcaccaac gctccaagtc tgacccctca
     3961 caactcatcc aaagtctcac tccacactct tgccaccgga ctggaaatgt ctctccttct
     4021 ggctgcctct cccaggctgc acggactcca ggagacagag cagggacaag caagtgcagg
     4081 aaagctggct gcatgtattt tgggactcca gaaaacaagg gcttttgcac tctatgtttc
     4141 atcgaataca gagaaaataa gcagtctgtt actgcctctg agaaagctgg ttccccggcc
     4201 cccaggttcc agaacaatgt cccgtgcctg ggcagggagt gcggcacact cggaagcacc
     4261 atgtttgaag ggtactgtca gaagtgtttc atcgaagctc agaaccagag attccatgaa
     4321 gcaagaagaa cggaagaaca gctgagatca agccagcata gagacatgcc tcgaactaca
     4381 caggtagcct caaggctgaa atgtgcccgg gcctcctgca agaacattct ggcctgtcgc
     4441 agtgaggaac tctgtatgga gtgccagcac ctaagccaac gagtaggttc tgtggcccac
     4501 cggggtgagc ccacgcctga agagccccct aaacagcgct gccgggcccc tgcttgtgat
     4561 cactttggca atgccaagtg taatggttac tgcaatgagt gctaccagtt caagcagatg
     4621 tatggcggat ccgactacaa agaccatgac ggtgattata aagatcatga catcgactac
     4681 aaggatgacg atgacaagta gcggccgcga aggatctgcg atcgctccgg tgcccgtcag
     4741 tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga
     4801 acgggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc
     4861 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt
     4921 ctttttcgca acgggtttgc cgccagaaca cagctgaagc ttcgaggggc tcgcatctct
     4981 ccttcacgcg cccgccgccc tacctgaggc cgccatccac gccggttgag tcgcgttctg
     5041 ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag tttaaagctc
     5101 aggtcgagac cgggcctttg tccggcgctc ccttggagcc tacctagact cagccggctc
     5161 tccacgcttt gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg
     5221 cgccgttaca gatccaagct gtgaccggcg cctacgctag atgaccgagt acaagcccac
     5281 ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt
     5341 cgccgactac cccgccacgc gccacaccgt cgacccggac cgccacatcg agcgggtcac
     5401 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     5461 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     5521 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     5581 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     5641 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     5701 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     5761 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     5821 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga gtcgacaatc aacctctgga
     5881 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     5941 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     6001 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     6061 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     6121 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     6181 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     6241 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
     6301 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     6361 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     6421 gacgagtcgg atctcccttt gggccgcctc cccgcctggt acctttaaga ccaatgactt
     6481 acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa
     6541 ttcactccca acgaaaataa gatctgcttt ttgcttgtac tgggtctctc tggttagacc
     6601 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     6661 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     6721 gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc
     6781 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt
     6841 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     6901 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     6961 tctggctcta gctatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     7021 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     7081 gaagtagtga ggaggctttt ttggaggcct agacttttgc agagacggcc caaattcgta
     7141 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     7201 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     7261 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     7321 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     7381 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     7441 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     7501 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     7561 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     7621 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     7681 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     7741 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     7801 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     7861 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     7921 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     7981 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     8041 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     8101 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     8161 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     8221 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     8281 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     8341 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     8401 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     8461 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     8521 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     8581 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     8641 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     8701 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     8761 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     8821 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     8881 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     8941 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     9001 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     9061 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     9121 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     9181 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     9241 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     9301 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     9361 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     9421 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     9481 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     9541 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     9601 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     9661 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     9721 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc tg
//
