LOCUS       V023774                 9397 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023774
VERSION     V023774
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..3511
                     /label="PTEN(NM_000314)"
                     /note="PTEN(NM_000314)"
                     /gene="PTEN"
     promoter        3556..3767
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3780..4048
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4077..4086
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4107..4769
                     /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             4839..4892
                     /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             4893..5492
                     /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    5493..6080
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6154..6387
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6459..6580
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6599..6734
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6772..6788)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6796..6812
                     /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(6820..6850)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6865..6886
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7174..7762)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7933..8793)
                     /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(8794..8898)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9372..9388
                     /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 ccatgacagc catcatcaaa gagatcgtta gcagaaacaa
     2341 aaggagatat caagaggatg gattcgactt agacttgacc tatatttatc caaacattat
     2401 tgctatggga tttcctgcag aaagacttga aggcgtatac aggaacaata ttgatgatgt
     2461 agtaaggttt ttggattcaa agcataaaaa ccattacaag atatacaatc tttgtgctga
     2521 aagacattat gacaccgcca aatttaattg cagagttgca caatatcctt ttgaagacca
     2581 taacccacca cagctagaac ttatcaaacc cttttgtgaa gatcttgacc aatggctaag
     2641 tgaagatgac aatcatgttg cagcaattca ctgtaaagct ggaaagggac gaactggtgt
     2701 aatgatatgt gcatatttat tacatcgggg caaattttta aaggcacaag aggccctaga
     2761 tttctatggg gaagtaagga ccagagacaa aaagggagta actattccca gtcagaggcg
     2821 ctatgtgtat tattatagct acctgttaaa gaatcatctg gattatagac cagtggcact
     2881 gttgtttcac aagatgatgt ttgaaactat tccaatgttc agtggcggaa cttgcaatcc
     2941 tcagtttgtg gtctgccagc taaaggtgaa gatatattcc tccaattcag gacccacacg
     3001 acgggaagac aagttcatgt actttgagtt ccctcagccg ttacctgtgt gtggtgatat
     3061 caaagtagag ttcttccaca aacagaacaa gatgctaaaa aaggacaaaa tgtttcactt
     3121 ttgggtaaat acattcttca taccaggacc agaggaaacc tcagaaaaag tagaaaatgg
     3181 aagtctatgt gatcaagaaa tcgatagcat ttgcagtata gagcgtgcag ataatgacaa
     3241 ggaatatcta gtacttactt taacaaaaaa tgatcttgac aaagcaaata aagacaaagc
     3301 caaccgatac ttttctccaa attttaaggt gaagctgtac ttcacaaaaa cagtagagga
     3361 gccgtcaaat ccagaggcta gcagttcaac ttctgtaaca ccagatgtta gtgacaatga
     3421 acctgatcat tatagatatt ctgacaccac tgactctgat ccagagaatg aaccttttga
     3481 tgaagatcag catacacaaa ttacaaaagt ctgagcggcc gcgaaggatc tgcgatcgct
     3541 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag
     3601 gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg
     3661 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag
     3721 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagctg aagcttcgag
     3781 gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat ccacgccggt
     3841 tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc gccgtctagg
     3901 taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg agcctaccta
     3961 gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctac gtctttgttt
     4021 cgttttctgt tctgcgccgt tacagatcca agctgtgacc ggcgcctacg ctagacgcca
     4081 ccatggagag cgacgagagc ggcctgcccg ccatggagat cgagtgccgc atcaccggca
     4141 ccctgaacgg cgtggagttc gagctggtgg gcggcggaga gggcaccccc aagcagggcc
     4201 gcatgaccaa caagatgaag agcaccaaag gcgccctgac cttcagcccc tacctgctga
     4261 gccacgtgat gggctacggc ttctaccact tcggcaccta ccccagcggc tacgagaacc
     4321 ccttcctgca cgccatcaac aacggcggct acaccaacac ccgcatcgag aagtacgagg
     4381 acggcggcgt gctgcacgtg agcttcagct accgctacga ggccggccgc gtgatcggcg
     4441 acttcaaggt ggtgggcacc ggcttccccg aggacagcgt gatcttcacc gacaagatca
     4501 tccgcagcaa cgccaccgtg gagcacctgc accccatggg cgataacgtg ctggtgggca
     4561 gcttcgcccg caccttcagc ctgcgcgacg gcggctacta cagcttcgtg gtggacagcc
     4621 acatgcactt caagagcgcc atccacccca gcatcctgca gaacgggggc cccatgttcg
     4681 ccttccgccg cgtggaggag ctgcacagca acaccgagct gggcatcgtg gagtaccagc
     4741 acgccttcaa gacccccatc gccttcgcca gatcccgcgc tcagtcgtcc aattctgccg
     4801 tggacggcac cgccggaccc ggctccaccg gatctcgcga gggcagagga agtcttctaa
     4861 catgcggtga cgtggaggag aatcccggcc ctatgaccga gtacaagccc acggtgcgcc
     4921 tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact
     4981 accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc
     5041 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
     5101 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
     5161 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
     5221 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg
     5281 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
     5341 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
     5401 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
     5461 cctggtgcat gacccgcaag cccggtgcct gaaatcaacc tctggattac aaaatttgtg
     5521 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     5581 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     5641 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     5701 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     5761 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     5821 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     5881 cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg
     5941 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     6001 tgctgccggc tctgcggcct cttccgcgtc tccgccttcg ccctcagacg agtcggatct
     6061 ccctttggcc gcctccccgc ctggtacctt taagaccaat gacttacaag gcagctgtag
     6121 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
     6181 aataagatct gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg
     6241 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     6301 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     6361 tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt
     6421 tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata
     6481 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     6541 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat
     6601 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     6661 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     6721 cttttttgga ggcctagact tttgcagaga cggcccaaat tcgtaatcat ggtcatagct
     6781 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     6841 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     6901 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     6961 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     7021 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     7081 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     7141 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     7201 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     7261 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     7321 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     7381 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     7441 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     7501 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     7561 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     7621 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     7681 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     7741 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7801 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     7861 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     7921 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7981 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     8041 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     8101 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     8161 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     8221 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     8281 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     8341 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     8401 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     8461 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     8521 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     8581 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     8641 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     8701 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     8761 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     8821 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     8881 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     8941 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     9001 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     9061 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     9121 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     9181 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     9241 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     9301 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     9361 gtcacgacgt tgtaaaacga cggccagtgc caagctg
//
