LOCUS       V025190                 9227 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025190
VERSION     V025190
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             2294..3655
                     /label="MYC(NM_002467)"
                     /note="MYC(NM_002467)"
                     /gene="MYC"
     CDS             3662..3727
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3770..3981
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3994..4262
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4290..5315
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /note="confers resistance to hygromycin"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /transl_table=1
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E*"
                     /label="HygR"
     misc_feature    5322..5910
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5984..6217
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6289..6410
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6429..6564
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6602..6618)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6626..6642
                     /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(6650..6680)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6695..6716
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7004..7592)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7763..8623)
                     /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(8624..8728)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9202..9218
                     /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 agagctagcc accatggatt tttttcgggt agtggaaaac cagcagcctc ccgcgacgat
     2341 gcccctcaac gttagcttca ccaacaggaa ctatgacctc gactacgact cggtgcagcc
     2401 gtatttctac tgcgacgagg aggagaactt ctaccagcag cagcagcaga gcgagctgca
     2461 gcccccggcg cccagcgagg atatctggaa gaaattcgag ctgctgccca ccccgcccct
     2521 gtcccctagc cgccgctccg ggctctgctc gccctcctac gttgcggtca cacccttctc
     2581 ccttcgggga gacaacgacg gcggtggcgg gagcttctcc acggccgacc agctggagat
     2641 ggtgaccgag ctgctgggag gagacatggt gaaccagagt ttcatctgcg acccggacga
     2701 cgagaccttc atcaaaaaca tcatcatcca ggactgtatg tggagcggct tctcggccgc
     2761 cgccaagctc gtctcagaga agctggcctc ctaccaggct gcgcgcaaag acagcggcag
     2821 cccgaacccc gcccgcggcc acagcgtctg ctccacctcc agcttgtacc tgcaggatct
     2881 gagcgccgcc gcctcagagt gcatcgaccc ctcggtggtc ttcccctacc ctctcaacga
     2941 cagcagctcg cccaagtcct gcgcctcgca agactccagc gccttctctc cgtcctcgga
     3001 ttctctgctc tcctcgacgg agtcctcccc gcagggcagc cccgagcccc tggtgctcca
     3061 tgaggagaca ccgcccacca ccagcagcga ctctgaggag gaacaagaag atgaggaaga
     3121 aatcgatgtt gtttctgtgg aaaagaggca ggctcctggc aaaaggtcag agtctggatc
     3181 accttctgct ggaggccaca gcaaacctcc tcacagccca ctggtcctca agaggtgcca
     3241 cgtctccaca catcagcaca actacgcagc gcctccctcc actcggaagg actatcctgc
     3301 tgccaagagg gtcaagttgg acagtgtcag agtcctgaga cagatcagca acaaccgaaa
     3361 atgcaccagc cccaggtcct cggacaccga ggagaatgtc aagaggcgaa cacacaacgt
     3421 cttggagcgc cagaggagga acgagctaaa acggagcttt tttgccctgc gtgaccagat
     3481 cccggagttg gaaaacaatg aaaaggcccc caaggtagtt atccttaaaa aagccacagc
     3541 atacatcctg tccgtccaag cagaggagca aaagctcatt tctgaagagg acttgttgcg
     3601 gaaacgacga gaacagttga aacacaaact tgaacagcta cggaactctt gtgcgctcga
     3661 ggactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga
     3721 tgacaagtag cggccgcaag gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc
     3781 acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac gggtgcctag
     3841 agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc
     3901 gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac
     3961 gggtttgccg ccagaacaca gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc
     4021 cgccgcccta cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc
     4081 tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg
     4141 ggcctttgtc cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc
     4201 ctgaccctgc ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga
     4261 tccaagctgt gaccggcgcc tacgtcgaca tgaaaaagcc tgaactcacc gcgacgtctg
     4321 tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga cctgatgcag ctctcggagg
     4381 gcgaagaatc tcgtgctttc agcttcgatg taggagggcg tggatatgtc ctgcgggtaa
     4441 atagctgcgc cgatggtttc tacaaagatc gttatgttta tcggcacttt gcatcggccg
     4501 cgctcccgat tccggaagtg cttgacattg gggaattcag cgagagcctg acctattgca
     4561 tctcccgccg tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg
     4621 ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc ggccgatctt agccagacga
     4681 gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata cactacatgg cgtgatttca
     4741 tatgcgcgat tgctgatccc catgtgtatc actggcaaac tgtgatggac gacaccgtca
     4801 gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac tgccccgaag
     4861 tccggcacct cgtgcacgcg gatttcggct ccaacaatgt cctgacggac aatggccgca
     4921 taacagcggt cattgactgg agcgaggcga tgttcgggga ttcccaatac gaggtcgcca
     4981 acatcttctt ctggaggccg tggttggctt gtatggagca gcagacgcgc tacttcgagc
     5041 ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc gtatatgctc cgcattggtc
     5101 ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct tgggcgcagg
     5161 gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca caaatcgccc
     5221 gcagaagcgc ggccgtctgg accgatggct gtgtagaagt actcgccgat agtggaaacc
     5281 gacgccccag cactcgtccg agggcaaagg aataggtcga caatcaacct ctggattaca
     5341 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     5401 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     5461 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     5521 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     5581 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     5641 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     5701 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga
     5761 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     5821 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     5881 gtcggatctc cctttgggcc gcctccccgc ctggtacctt taagaccaat gacttacaag
     5941 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac
     6001 tcccaacgaa aataagatct gctttttgct tgtactgggt ctctctggtt agaccagatc
     6061 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     6121 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     6181 ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt
     6241 attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt
     6301 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     6361 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     6421 ctctagctat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     6481 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     6541 agtgaggagg cttttttgga ggcctagact tttgcagaga cggcccaaat tcgtaatcat
     6601 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     6661 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     6721 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     6781 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     6841 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6901 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6961 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     7021 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     7081 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     7141 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     7201 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     7261 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     7321 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     7381 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     7441 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     7501 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7561 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7621 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7681 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7741 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7801 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     7861 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     7921 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7981 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     8041 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     8101 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     8161 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     8221 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     8281 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     8341 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     8401 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     8461 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     8521 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     8581 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8641 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8701 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     8761 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     8821 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     8881 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     8941 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     9001 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     9061 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     9121 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     9181 ggttttccca gtcacgacgt tgtaaaacga cggccagtgc caagctg
//
