LOCUS       V024198                 9341 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024198
VERSION     V024198
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             2317..3384
                     /label="Ube2z(NM_172300)"
                     /note="Ube2z(NM_172300)"
                     /gene="Ube2z"
     CDS             3391..3456
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3500..3711
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3724..3992
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4021..4030
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4051..4713
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /transl_table=1
                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
                     /label="CopGFP"
     CDS             4783..4836
                     /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             4837..5436
                     /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    5437..6024
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6098..6331
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6403..6524
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6543..6678
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6716..6732)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6740..6756
                     /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(6764..6794)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6809..6830
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7118..7706)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7877..8737)
                     /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(8738..8842)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9316..9332
                     /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 ctgatacgaa ctcggaattc gccaccatgg cggagagtcc gactgaggag
     2341 gcggcgacgg cgacggctgg cgccggggcg gcgggcccag ggtcgagtgg tgttgccggt
     2401 gtggtcggcg ttagcgggag cggcggcggg ttcgggccgc cttttctgcc ggatgtgtgg
     2461 gcggcggcgg cggcggcggg cggggccggg ggaccgggga gcggcctggc tccgctgcca
     2521 ggactcccgc cctcggccgc tgcccacggg gccgcgctgc ttagccattg ggaccccacg
     2581 ctcagctcgg actgggacgg cgagcgaacc gcgccacagt gtctacttcg gattaagcgg
     2641 gatatcatgt ccatttataa ggagcctcct ccaggaatgt tcgttgtgcc cgacactgtt
     2701 gacatgacta agattcatgc tttgatcaca ggcccatttg acactcctta tgaagggggt
     2761 ttcttcctgt tcgtctttcg gtgtccacct gactatccca tccacccacc tcgggtcaaa
     2821 ctgatgacaa cgggcaataa cacagtgagg tttaacccca acttctaccg caatgggaaa
     2881 gtctgcttga gtattctagg tacgtggact ggccctgcct ggagcccagc ccagagcatc
     2941 tcttctgtgc ttatctccat ccagtccctg atgactgaga atccctacca caacgaacca
     3001 ggctttgaac aggaaagaca tccaggagac agcaaaaact ataatgaatg tatccggcac
     3061 gagaccatca gggttgcagt ctgtgacatg atggagggca agtgcccctg ccctgaaccc
     3121 ctacggggag tgatggagaa gtccttcctg gagtattatg acttctatga ggtggcctgc
     3181 aaagatcgcc tgcatcttca aggccaaact atgcaggacc cttttggaga gaagcggggc
     3241 cactttgact accagtccct cttgatgcgc ctcggactga ttcgacagaa agtgctggaa
     3301 aggctccata atgaaaatgc tgaaatggac tctgatagca gctcctctgg gacagagaca
     3361 gaccttcatg ggagcctgag ggtcggatcc gactacaaag accatgacgg tgattataaa
     3421 gatcatgaca tcgactacaa ggatgacgat gacaagtagc ggccgcgaag gatctgcgat
     3481 cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
     3541 ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
     3601 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
     3661 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt
     3721 cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc
     3781 cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc
     3841 taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta
     3901 cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt
     3961 gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagac
     4021 gccaccatgg agagcgacga gagcggcctg cccgccatgg agatcgagtg ccgcatcacc
     4081 ggcaccctga acggcgtgga gttcgagctg gtgggcggcg gagagggcac ccccaagcag
     4141 ggccgcatga ccaacaagat gaagagcacc aaaggcgccc tgaccttcag cccctacctg
     4201 ctgagccacg tgatgggcta cggcttctac cacttcggca cctaccccag cggctacgag
     4261 aaccccttcc tgcacgccat caacaacggc ggctacacca acacccgcat cgagaagtac
     4321 gaggacggcg gcgtgctgca cgtgagcttc agctaccgct acgaggccgg ccgcgtgatc
     4381 ggcgacttca aggtggtggg caccggcttc cccgaggaca gcgtgatctt caccgacaag
     4441 atcatccgca gcaacgccac cgtggagcac ctgcacccca tgggcgataa cgtgctggtg
     4501 ggcagcttcg cccgcacctt cagcctgcgc gacggcggct actacagctt cgtggtggac
     4561 agccacatgc acttcaagag cgccatccac cccagcatcc tgcagaacgg gggccccatg
     4621 ttcgccttcc gccgcgtgga ggagctgcac agcaacaccg agctgggcat cgtggagtac
     4681 cagcacgcct tcaagacccc catcgccttc gccagatccc gcgctcagtc gtccaattct
     4741 gccgtggacg gcaccgccgg acccggctcc accggatctc gcgagggcag aggaagtctt
     4801 ctaacatgcg gtgacgtgga ggagaatccc ggccctatga ccgagtacaa gcccacggtg
     4861 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc
     4921 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag
     4981 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac
     5041 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc
     5101 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag
     5161 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc
     5221 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg
     5281 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc
     5341 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg
     5401 cgcacctggt gcatgacccg caagcccggt gcctgaaatc aacctctgga ttacaaaatt
     5461 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     5521 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     5581 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     5641 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     5701 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     5761 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     5821 ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg
     5881 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     5941 ggcctgctgc cggctctgcg gcctcttccg cgtctccgcc ttcgccctca gacgagtcgg
     6001 atctcccttt ggccgcctcc ccgcctggta cctttaagac caatgactta caaggcagct
     6061 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     6121 cgaaaataag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
     6181 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     6241 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     6301 cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag
     6361 tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct
     6421 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     6481 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag
     6541 ctatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     6601 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     6661 gaggcttttt tggaggccta gacttttgca gagacggccc aaattcgtaa tcatggtcat
     6721 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     6781 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     6841 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     6901 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     6961 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     7021 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     7081 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     7141 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     7201 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     7261 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     7321 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     7381 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     7441 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     7501 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     7561 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     7621 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     7681 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     7741 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     7801 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     7861 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     7921 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7981 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     8041 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     8101 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     8161 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     8221 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     8281 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     8341 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     8401 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     8461 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     8521 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     8581 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     8641 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     8701 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     8761 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     8821 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     8881 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     8941 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     9001 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     9061 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     9121 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt
     9181 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     9241 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     9301 cccagtcacg acgttgtaaa acgacggcca gtgccaagct g
//
