LOCUS       V032046                 9207 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032046
VERSION     V032046
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             2293..3288
                     /label="Ldha(NM_010699)"
                     /note="Ldha(NM_010699)"
                     /gene="Ldha"
     CDS             3292..3321
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     promoter        3366..3577
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3590..3858
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3887..3896
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3917..4579
                     /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             4649..4702
                     /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             4703..5302
                     /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    5303..5890
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5964..6197
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6269..6390
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6409..6544
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6582..6598)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6606..6622
                     /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(6630..6660)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6675..6696
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6984..7572)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7743..8603)
                     /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(8604..8708)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9182..9198
                     /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 gaatggcaac cctcaaggac cagctgattg tgaatcttct taaggaagag
     2341 caggctcccc agaacaagat tacagttgtt ggggttggtg ctgttggcat ggcttgtgcc
     2401 atcagtatct taatgaagga cttggcggat gagcttgccc ttgttgacgt catggaagac
     2461 aaactcaagg gcgagatgat ggatctccag catggcagcc tcttccttaa aacaccaaaa
     2521 attgtctcca gcaaagacta ctgtgtaact gcgaactcca agctggtcat tatcaccgcg
     2581 ggggcccgtc agcaagaggg ggagagccgg ctcaacctgg tccagcgaaa cgtgaacatc
     2641 ttcaagttca tcattcccaa cattgtcaag tacagtccac actgcaagct gctgatcgtc
     2701 tccaatccag tggatatctt gacctacgtg gcttggaaaa tcagtggctt tcccaaaaac
     2761 cgagtaattg gaagtggttg caatctggat tcagcgcggt tccgttacct gatgggagag
     2821 aggctggggg ttcacgcgct gagctgtcac ggctgggtcc tgggagaaca tggcgactcc
     2881 agtgtgcctg tgtggagtgg tgtgaatgtt gccggcgtct ccctgaagtc tcttaaccca
     2941 gaactgggca ctgacgcaga caaggagcag tggaaggagg ttcacaagca ggtggtggac
     3001 agtgcctacg aggtgatcaa gctgaaaggt tacacatcct gggccattgg cctctctgtg
     3061 gcagacttgg ctgagagcat aatgaagaac cttaggcggg tgcatcccat ttccaccatg
     3121 attaagggtc tctatggaat caatgaggat gtcttcctca gtgtcccatg tatcctggga
     3181 caaaatggaa tctcggatgt tgtgaaggtg acactgactc ctgaggaaga ggcccgcctg
     3241 aagaagagcg cagacaccct ctggggaatc cagaaggagc tgcagttctc cgaacaaaaa
     3301 ctcatctcag aagaggatct gtgagcggcc gcgaaggatc tgcgatcgct ccggtgcccg
     3361 tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa
     3421 ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg
     3481 gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa
     3541 cgttcttttt cgcaacgggt ttgccgccag aacacagctg aagcttcgag gggctcgcat
     3601 ctctccttca cgcgcccgcc gccctacctg aggccgccat ccacgccggt tgagtcgcgt
     3661 tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa
     3721 gctcaggtcg agaccgggcc tttgtccggc gctcccttgg agcctaccta gactcagccg
     3781 gctctccacg ctttgcctga ccctgcttgc tcaactctac gtctttgttt cgttttctgt
     3841 tctgcgccgt tacagatcca agctgtgacc ggcgcctacg ctagacgcca ccatggagag
     3901 cgacgagagc ggcctgcccg ccatggagat cgagtgccgc atcaccggca ccctgaacgg
     3961 cgtggagttc gagctggtgg gcggcggaga gggcaccccc aagcagggcc gcatgaccaa
     4021 caagatgaag agcaccaaag gcgccctgac cttcagcccc tacctgctga gccacgtgat
     4081 gggctacggc ttctaccact tcggcaccta ccccagcggc tacgagaacc ccttcctgca
     4141 cgccatcaac aacggcggct acaccaacac ccgcatcgag aagtacgagg acggcggcgt
     4201 gctgcacgtg agcttcagct accgctacga ggccggccgc gtgatcggcg acttcaaggt
     4261 ggtgggcacc ggcttccccg aggacagcgt gatcttcacc gacaagatca tccgcagcaa
     4321 cgccaccgtg gagcacctgc accccatggg cgataacgtg ctggtgggca gcttcgcccg
     4381 caccttcagc ctgcgcgacg gcggctacta cagcttcgtg gtggacagcc acatgcactt
     4441 caagagcgcc atccacccca gcatcctgca gaacgggggc cccatgttcg ccttccgccg
     4501 cgtggaggag ctgcacagca acaccgagct gggcatcgtg gagtaccagc acgccttcaa
     4561 gacccccatc gccttcgcca gatcccgcgc tcagtcgtcc aattctgccg tggacggcac
     4621 cgccggaccc ggctccaccg gatctcgcga gggcagagga agtcttctaa catgcggtga
     4681 cgtggaggag aatcccggcc ctatgaccga gtacaagccc acggtgcgcc tcgccacccg
     4741 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     4801 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     4861 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     4921 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     4981 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     5041 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     5101 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     5161 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     5221 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     5281 gacccgcaag cccggtgcct gaaatcaacc tctggattac aaaatttgtg aaagattgac
     5341 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     5401 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     5461 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     5521 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     5581 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     5641 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     5701 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     5761 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     5821 tctgcggcct cttccgcgtc tccgccttcg ccctcagacg agtcggatct ccctttggcc
     5881 gcctccccgc ctggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca
     5941 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa aataagatct
     6001 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     6061 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     6121 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     6181 gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc
     6241 aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa
     6301 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     6361 tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta
     6421 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     6481 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     6541 ggcctagact tttgcagaga cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg
     6601 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     6661 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     6721 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     6781 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6841 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6901 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6961 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     7021 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     7081 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     7141 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     7201 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     7261 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     7321 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     7381 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     7441 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     7501 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     7561 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     7621 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     7681 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     7741 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     7801 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     7861 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     7921 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     7981 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     8041 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     8101 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     8161 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     8221 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     8281 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     8341 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     8401 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     8461 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     8521 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     8581 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     8641 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     8701 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     8761 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
     8821 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     8881 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     8941 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa
     9001 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg
     9061 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     9121 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     9181 tgtaaaacga cggccagtgc caagctg
//
