LOCUS       V031747                 9157 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031747
VERSION     V031747
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..3472
                     /label="Trp53(NM_011640)"
                     /note="Trp53(NM_011640)"
                     /gene="Trp53"
     CDS             3479..3505
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     promoter        3550..3761
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3774..4042
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4077..4784
                     /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             4794..4847
                     /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             4848..5246
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /note="confers resistance to blasticidin"
                     /product="blasticidin S deaminase"
                     /transl_table=1
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG*"
                     /label="BSD"
     misc_feature    5253..5840
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5914..6147
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6219..6340
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6359..6494
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6532..6548)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6556..6572
                     /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(6580..6610)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6625..6646
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6934..7522)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7693..8553)
                     /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(8554..8658)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9132..9148
                     /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 ccatgactgc catggaggag tcacagtcgg atatcagcct
     2341 cgagctccct ctgagccagg agacattttc aggcttatgg aaactacttc ctccagaaga
     2401 tatcctgcca tcacctcact gcatggacga tctgttgctg ccccaggatg ttgaggagtt
     2461 ttttgaaggc ccaagtgaag ccctccgagt gtcaggagct cctgcagcac aggaccctgt
     2521 caccgagacc cctgggccag tggcccctgc cccagccact ccatggcccc tgtcatcttt
     2581 tgtcccttct caaaaaactt accagggcaa ctatggcttc cacctgggct tcctgcagtc
     2641 tgggacagcc aagtctgtta tgtgcacgta ctctcctccc ctcaataagc tattctgcca
     2701 gctggcgaag acgtgccctg tgcagttgtg ggtcagcgcc acacctccag ctgggagccg
     2761 tgtccgcgcc atggccatct acaagaagtc acagcacatg acggaggtcg tgagacgctg
     2821 cccccaccat gagcgctgct ccgatggtga tggcctggct cctccccagc atcttatccg
     2881 ggtggaagga aatttgtatc ccgagtatct ggaagacagg cagacttttc gccacagcgt
     2941 ggtggtacct tatgagccac ccgaggccgg ctctgagtat accaccatcc actacaagta
     3001 catgtgtaat agctcctgca tggggggcat gaaccgccga cctatcctta ccatcatcac
     3061 actggaagac tccagtggga accttctggg acgggacagc tttgaggttc gtgtttgtgc
     3121 ctgccctggg agagaccgcc gtacagaaga agaaaatttc cgcaaaaagg aagtcctttg
     3181 ccctgaactg cccccaggga gcgcaaagag agcgctgccc acctgcacaa gcgcctctcc
     3241 cccgcaaaag aaaaaaccac ttgatggaga gtatttcacc ctcaagatcc gcgggcgtaa
     3301 acgcttcgag atgttccggg agctgaatga ggccttagag ttaaaggatg cccatgctac
     3361 agaggagtct ggagacagca gggctcactc cagctacctg aagaccaaga agggccagtc
     3421 tacttcccgc cataaaaaaa caatggtcaa gaaagtgggg cctgactcag acggatccta
     3481 cccatacgac gtaccagatt acgcttgagc ggccgcgaag gatctgcgat cgctccggtg
     3541 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
     3601 gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
     3661 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
     3721 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt cgaggggctc
     3781 gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc cggttgagtc
     3841 gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt
     3901 taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta cctagactca
     3961 gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt gtttcgtttt
     4021 ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagac gccaccatgg
     4081 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc
     4141 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     4201 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg
     4261 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg
     4321 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
     4381 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg
     4441 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc
     4501 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc
     4561 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg
     4621 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca
     4681 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg
     4741 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagggatct cgcgagggca
     4801 gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc cggccctatg gccaagcctt
     4861 tgtctcaaga agaatccacc ctcattgaaa gagcaacggc tacaatcaac agcatcccca
     4921 tctctgaaga ctacagcgtc gccagcgcag ctctctctag cgacggccgc atcttcactg
     4981 gtgtcaatgt atatcatttt actgggggac cttgtgcaga actcgtggtg ctgggcactg
     5041 ctgctgctgc ggcagctggc aacctgactt gtatcgtcgc gatcggaaat gagaacaggg
     5101 gcatcttgag cccctgcgga cggtgccgac aggtgcttct cgatctgcat cctgggatca
     5161 aagccatagt gaaggacagt gatggacagc cgacggcagt tgggattcgt gaattgctgc
     5221 cctctggtta tgtgtgggag ggctaagtcg acaatcaacc tctggattac aaaatttgtg
     5281 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     5341 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     5401 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     5461 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     5521 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     5581 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     5641 cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg
     5701 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     5761 tgctgccggc tctgcggcct cttccgcgtc tccgccttcg ccctcagacg agtcggatct
     5821 ccctttggcc gcctccccgc ctggtacctt taagaccaat gacttacaag gcagctgtag
     5881 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
     5941 gataagatct gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg
     6001 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     6061 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     6121 tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt
     6181 tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata
     6241 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     6301 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat
     6361 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     6421 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     6481 cttttttgga ggcctagact tttgcagaga cggcccaaat tcgtaatcat ggtcatagct
     6541 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     6601 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     6661 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     6721 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     6781 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6841 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6901 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6961 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     7021 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     7081 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     7141 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     7201 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     7261 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     7321 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     7381 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     7441 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     7501 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7561 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     7621 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     7681 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7741 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7801 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7861 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7921 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7981 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     8041 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     8101 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     8161 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     8221 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     8281 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     8341 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     8401 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     8461 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     8521 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     8581 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     8641 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     8701 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     8761 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     8821 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     8881 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     8941 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     9001 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     9061 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     9121 gtcacgacgt tgtaaaacga cggccagtgc caagctg
//
