LOCUS       V044457                 9291 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044457
VERSION     V044457
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..3373
                     /label="MAPK3(NM_001040056)"
                     /note="MAPK3(NM_001040056)"
                     /gene="MAPK3"
     CDS             3380..3445
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3489..3700
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3713..3981
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4016..4723
                     /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             4733..4786
                     /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             4787..5386
                     /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    5387..5974
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6048..6281
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6353..6474
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6493..6628
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6666..6682)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6690..6706
                     /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(6714..6744)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6759..6780
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7068..7656)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7827..8687)
                     /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(8688..8792)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9266..9282
                     /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 ccatggcggc ggcggcggct caggggggcg ggggcgggga
     2341 gccccgtaga accgaggggg tcggcccggg ggtcccgggg gaggtggaga tggtgaaggg
     2401 gcagccgttc gacgtgggcc cgcgctacac gcagttgcag tacatcggcg agggcgcgta
     2461 cggcatggtc agctcggcct atgaccacgt gcgcaagact cgcgtggcca tcaagaagat
     2521 cagccccttc gaacatcaga cctactgcca gcgcacgctc cgggagatcc agatcctgct
     2581 gcgcttccgc catgagaatg tcatcggcat ccgagacatt ctgcgggcgt ccaccctgga
     2641 agccatgaga gatgtctaca ttgtgcagga cctgatggag actgacctgt acaagttgct
     2701 gaaaagccag cagctgagca atgaccatat ctgctacttc ctctaccaga tcctgcgggg
     2761 cctcaagtac atccactccg ccaacgtgct ccaccgagat ctaaagccct ccaacctgct
     2821 catcaacacc acctgcgacc ttaagatttg tgatttcggc ctggcccgga ttgccgatcc
     2881 tgagcatgac cacaccggct tcctgacgga gtatgtggct acgcgctggt accgggcccc
     2941 agagatcatg ctgaactcca agggctatac caagtccatc gacatctggt ctgtgggctg
     3001 cattctggct gagatgctct ctaaccggcc catcttccct ggcaagcact acctggatca
     3061 gctcaaccac attctgggca tcctgggctc cccatcccag gaggacctga attgtatcat
     3121 caacatgaag gcccgaaact acctacagtc tctgccctcc aagaccaagg tggcttgggc
     3181 caagcttttc cccaagtcag actccaaagc ccttgacctg ctggaccgga tgttaacctt
     3241 taaccccaat aaacggatca cagtggagga agcgctggct cacccctacc tggagcagta
     3301 ctatgacccg acggatgagg tgggccagtc cccagcagca gtggggctgg gggcagggga
     3361 gcaggggggc acgggatccg actacaaaga ccatgacggt gattataaag atcatgacat
     3421 cgactacaag gatgacgatg acaagtagcg gccgcgaagg atctgcgatc gctccggtgc
     3481 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg
     3541 caattgaacg ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta
     3601 ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt
     3661 gaacgttctt tttcgcaacg ggtttgccgc cagaacacag ctgaagcttc gaggggctcg
     3721 catctctcct tcacgcgccc gccgccctac ctgaggccgc catccacgcc ggttgagtcg
     3781 cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt
     3841 aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct tggagcctac ctagactcag
     3901 ccggctctcc acgctttgcc tgaccctgct tgctcaactc tacgtctttg tttcgttttc
     3961 tgttctgcgc cgttacagat ccaagctgtg accggcgcct acgctagacg ccaccatggt
     4021 gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca
     4081 catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     4141 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc
     4201 ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc
     4261 cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
     4321 gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga
     4381 gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca
     4441 gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct
     4501 gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt
     4561 caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat
     4621 caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc
     4681 cgagggccgc cactccaccg gcggcatgga cgagctgtac aagggatctc gcgagggcag
     4741 aggaagtctt ctaacatgcg gtgacgtgga ggagaatccc ggccctatga ccgagtacaa
     4801 gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc
     4861 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg
     4921 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     4981 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     5041 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     5101 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     5161 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     5221 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc
     5281 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     5341 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgaaatc aacctctgga
     5401 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     5461 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     5521 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     5581 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     5641 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     5701 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     5761 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     5821 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     5881 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtctccgcc ttcgccctca
     5941 gacgagtcgg atctcccttt ggccgcctcc ccgcctggta cctttaagac caatgactta
     6001 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat
     6061 tcactcccaa cgaaaataag atctgctttt tgcttgtact gggtctctct ggttagacca
     6121 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     6181 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     6241 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct
     6301 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt
     6361 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     6421 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     6481 ctggctctag ctatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     6541 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     6601 aagtagtgag gaggcttttt tggaggccta gacttttgca gagacggccc aaattcgtaa
     6661 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     6721 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     6781 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     6841 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     6901 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     6961 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     7021 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     7081 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     7141 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     7201 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     7261 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     7321 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     7381 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     7441 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     7501 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     7561 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     7621 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     7681 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     7741 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     7801 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7861 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     7921 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     7981 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     8041 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     8101 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     8161 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     8221 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     8281 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     8341 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     8401 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     8461 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     8521 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     8581 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     8641 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     8701 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     8761 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     8821 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     8881 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     8941 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     9001 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     9061 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     9121 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     9181 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     9241 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagct g
//
