LOCUS       V043269                 9504 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043269
VERSION     V043269
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..1921
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2011..2214
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2303..3586
                     /label="Hdac3(NM_010411)"
                     /note="Hdac3(NM_010411)"
                     /gene="Hdac3"
     CDS             3593..3658
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3702..3913
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3926..4194
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4229..4936
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
                     /label="mCherry"
     CDS             4946..4999
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from&nbsp;Thosea asigna virus capsid
                     protein"
                     /protein_id=""
                     /transl_table=1
                     /translation="EGRGSLLTCGDVEENPGP"
                     /label="T2A"
     CDS             5000..5599
                     /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    5600..6187
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6261..6494
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6566..6687
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6706..6841
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6879..6895)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6903..6919
                     /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(6927..6957)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6972..6993
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7281..7869)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8040..8900)
                     /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(8901..9005)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9479..9495
                     /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 attacaaaaa ttcaaaattt
     1921 tatcgatact agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta
     2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc
     2281 tagagctagc gaattcgcca ccatggccaa gaccgtggcg tatttctacg accccgatgt
     2341 gggcaacttc cactatggag ctggacaccc aatgaaacct catcgcctgg cattgactca
     2401 tagcctagtc ctgcattatg gtctctataa gaagatgatc gtcttcaagc cttaccaggc
     2461 ctcccagcat gacatgtgcc gcttccattc tgaggactac atcgacttcc tgcagagagt
     2521 cagccccacc aatatgcagg gtttcaccaa gagccttaat gccttcaacg tgggtgatga
     2581 ctgcccagtg tttccaggac ttttcgagtt ctgctcccgt tacacaggcg catctctgca
     2641 aggagcaaca cagctaaaca acaagatctg tgatattgcc atcaactggg ccggtggtct
     2701 acatcatgcc aagaaatttg aggcctctgg cttctgctat gtcaatgaca tagtaattgg
     2761 tatcctggag ctgcttaagt accaccctcg ggtgctctac attgatatcg acatccacca
     2821 tggtgacggg gttcaggaag ccttctacct cactgaccgg gtcatgactg tgtccttcca
     2881 caaatacgga aattacttct ttcctggaac aggtgacatg tatgaagttg gagcagagag
     2941 tggccgctac tattgtctca atgtgccctt acgagatggc attgatgacc agagttacaa
     3001 gcaccttttc cagccagtca tcagccaggt ggtggacttc taccagccga cgtgcatcgt
     3061 gctccagtgt ggcgctgact ccctgggctg tgatcgatta ggctgcttca atctcagcat
     3121 tcgaggacat ggggaatgtg ttgaatatgt caagagtttc aatatccctc tcctggtact
     3181 gggaggtggt ggctacactg tccgaaatgt tgcccggtgt tggacatatg aaacatctct
     3241 gctggtagaa gaggccatta gtgaggaact tccctatagt gaatacttcg agtactttgc
     3301 cccagatttc acactccatc cagatgtcag cacccgcatc gagaatcaga actcacgcca
     3361 gtatctggac cagatccgcc agacaatctt tgaaaacttg aagatgctga accatgcacc
     3421 cagtgtccag attcatgatg tcccggcaga cctcctgacg tatgacagga ctgacgaggc
     3481 cgacgctgaa gagagaggtc ccgaggagaa ctacagcagg ccagaagcac ccaatgagtt
     3541 ctatgatggc gaccatgaca acgacaagga aagtgatgtg gagattggat ccgactacaa
     3601 agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta
     3661 gcggccgcga aggatctgcg atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc
     3721 cacagtcccc gagaagttgg ggggaggggt cggcaattga acgggtgcct agagaaggtg
     3781 gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg
     3841 gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc
     3901 cgccagaaca cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc
     3961 tacctgaggc cgccatccac gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc
     4021 ctcctgaact gcgtccgccg tctaggtaag tttaaagctc aggtcgagac cgggcctttg
     4081 tccggcgctc ccttggagcc tacctagact cagccggctc tccacgcttt gcctgaccct
     4141 gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca gatccaagct
     4201 gtgaccggcg cctacgctag acgccaccat ggtgagcaag ggcgaggagg ataacatggc
     4261 catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga
     4321 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     4381 gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat
     4441 gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc
     4501 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac
     4561 cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg
     4621 caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc
     4681 ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa
     4741 gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc
     4801 cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga
     4861 ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat
     4921 ggacgagctg tacaagggat ctcgcgaggg cagaggaagt cttctaacat gcggtgacgt
     4981 ggaggagaat cccggcccta tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
     5041 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
     5101 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
     5161 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
     5221 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
     5281 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
     5341 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca
     5401 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
     5461 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
     5521 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
     5581 ccgcaagccc ggtgcctgaa atcaacctct ggattacaaa atttgtgaaa gattgactgg
     5641 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     5701 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     5761 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     5821 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     5881 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     5941 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     6001 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     6061 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     6121 gcggcctctt ccgcgtctcc gccttcgccc tcagacgagt cggatctccc tttggccgcc
     6181 tccccgcctg gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     6241 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaaaat aagatctgct
     6301 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     6361 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     6421 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     6481 gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa
     6541 gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata
     6601 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     6661 tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact
     6721 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     6781 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     6841 ctagactttt gcagagacgg cccaaattcg taatcatggt catagctgtt tcctgtgtga
     6901 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     6961 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     7021 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     7081 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     7141 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     7201 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     7261 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     7321 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     7381 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     7441 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     7501 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     7561 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     7621 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     7681 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     7741 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     7801 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     7861 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     7921 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     7981 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     8041 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     8101 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     8161 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     8221 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     8281 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     8341 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     8401 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     8461 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     8521 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     8581 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     8641 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     8701 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     8761 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     8821 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     8881 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     8941 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     9001 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     9061 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     9121 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     9181 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     9241 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
     9301 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc
     9361 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     9421 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     9481 aaaacgacgg ccagtgccaa gctg
//
