LOCUS       V025434                 9091 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025434
VERSION     V025434
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             2291..3169
                     /label="CASP6(NM_001226)"
                     /note="CASP6(NM_001226)"
                     /gene="CASP6"
     CDS             3176..3199
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHHHH"
                     /label="8xHis"
     promoter        3250..3461
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3474..3742
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3771..3780
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3801..4463
                     /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             4533..4586
                     /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             4587..5186
                     /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    5187..5774
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5848..6081
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6153..6274
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6293..6428
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6466..6482)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6490..6506
                     /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(6514..6544)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6559..6580
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6868..7456)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7627..8487)
                     /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(8488..8592)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9066..9082
                     /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 tagagccacc atgagctcgg cctcggggct ccgcaggggg cacccggcag gtggggaaga
     2341 aaacatgaca gaaacagatg ccttctataa aagagaaatg tttgatccgg cagaaaagta
     2401 caaaatggac cacaggagga gaggaattgc tttaatcttc aatcatgaga ggttcttttg
     2461 gcacttaaca ctgccagaaa ggcggggcac ctgcgcagat agagacaatc ttacccgcag
     2521 gttttcagat ctaggatttg aagtgaaatg ctttaatgat cttaaagcag aagaactact
     2581 gctcaaaatt catgaggtgt caactgttag ccacgcagat gccgattgct ttgtgtgtgt
     2641 cttcctgagc catggcgaag gcaatcacat ttatgcatat gatgctaaaa tcgaaattca
     2701 gacattaact ggcttgttca aaggagacaa gtgtcacagc ctggttggaa aacccaagat
     2761 atttatcatt caggcatgtc ggggaaacca gcacgatgtg ccagtcattc ctttggatgt
     2821 agtagataat cagacagaga agttggacac caacataact gaggtggatg cagcctccgt
     2881 ttacacgctg cctgctggag ctgacttcct catgtgttac tctgttgcag aaggatatta
     2941 ttctcaccgg gaaactgtga acggctcatg gtacattcaa gatttgtgtg agatgttggg
     3001 aaaatatggc tcctccttag agttcacaga actcctcaca ctggtgaaca ggaaagtttc
     3061 tcagcgccga gtggactttt gcaaagaccc aagtgcaatt ggaaagaagc aggttccctg
     3121 ttttgcctca atgctaacta aaaagctgca tttctttcca aaatctaatg aattccacca
     3181 tcaccatcac catcaccatt gaggatccgc ggccgcgaag gatctgcgat cgctccggtg
     3241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
     3301 gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
     3361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
     3421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt cgaggggctc
     3481 gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc cggttgagtc
     3541 gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt
     3601 taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta cctagactca
     3661 gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt gtttcgtttt
     3721 ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagac gccaccatgg
     3781 agagcgacga gagcggcctg cccgccatgg agatcgagtg ccgcatcacc ggcaccctga
     3841 acggcgtgga gttcgagctg gtgggcggcg gagagggcac ccccaagcag ggccgcatga
     3901 ccaacaagat gaagagcacc aaaggcgccc tgaccttcag cccctacctg ctgagccacg
     3961 tgatgggcta cggcttctac cacttcggca cctaccccag cggctacgag aaccccttcc
     4021 tgcacgccat caacaacggc ggctacacca acacccgcat cgagaagtac gaggacggcg
     4081 gcgtgctgca cgtgagcttc agctaccgct acgaggccgg ccgcgtgatc ggcgacttca
     4141 aggtggtggg caccggcttc cccgaggaca gcgtgatctt caccgacaag atcatccgca
     4201 gcaacgccac cgtggagcac ctgcacccca tgggcgataa cgtgctggtg ggcagcttcg
     4261 cccgcacctt cagcctgcgc gacggcggct actacagctt cgtggtggac agccacatgc
     4321 acttcaagag cgccatccac cccagcatcc tgcagaacgg gggccccatg ttcgccttcc
     4381 gccgcgtgga ggagctgcac agcaacaccg agctgggcat cgtggagtac cagcacgcct
     4441 tcaagacccc catcgccttc gccagatccc gcgctcagtc gtccaattct gccgtggacg
     4501 gcaccgccgg acccggctcc accggatctc gcgagggcag aggaagtctt ctaacatgcg
     4561 gtgacgtgga ggagaatccc ggccctatga ccgagtacaa gcccacggtg cgcctcgcca
     4621 cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg
     4681 ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac
     4741 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg
     4801 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg
     4861 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc
     4921 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc
     4981 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg
     5041 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg
     5101 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt
     5161 gcatgacccg caagcccggt gcctgaaatc aacctctgga ttacaaaatt tgtgaaagat
     5221 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     5281 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     5341 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     5401 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     5461 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     5521 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     5581 agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     5641 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     5701 cggctctgcg gcctcttccg cgtctccgcc ttcgccctca gacgagtcgg atctcccttt
     5761 ggccgcctcc ccgcctggta cctttaagac caatgactta caaggcagct gtagatctta
     5821 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaaaataag
     5881 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     5941 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     6001 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     6061 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac
     6121 ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt
     6181 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     6241 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc
     6301 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     6361 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     6421 tggaggccta gacttttgca gagacggccc aaattcgtaa tcatggtcat agctgtttcc
     6481 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     6541 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     6601 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     6661 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     6721 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6781 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     6841 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6901 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6961 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     7021 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     7081 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     7141 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     7201 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7261 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     7321 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     7381 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     7441 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     7501 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7561 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     7621 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     7681 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     7741 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     7801 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     7861 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     7921 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     7981 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     8041 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     8101 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     8161 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     8221 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     8281 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     8341 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     8401 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     8461 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     8521 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     8581 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     8641 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     8701 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     8761 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     8821 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     8881 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     8941 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     9001 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     9061 acgttgtaaa acgacggcca gtgccaagct g
//
