LOCUS       V022326                 9550 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022326
VERSION     V022326
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..3664
                     /label="DAPK3(NM_001348)"
                     /note="DAPK3(NM_001348)"
                     /gene="DAPK3"
     promoter        3709..3920
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3933..4201
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4230..4239
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4260..4922
                     /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             4992..5045
                     /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             5046..5645
                     /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    5646..6233
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6307..6540
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6612..6733
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6752..6887
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6925..6941)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6949..6965
                     /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(6973..7003)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7018..7039
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7327..7915)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8086..8946)
                     /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(8947..9051)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9525..9541
                     /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 ccatgtccac gttcaggcag gaggacgtgg aggaccatta
     2341 tgagatgggg gaggagctgg gcagcggcca gtttgcgatc gtgcggaagt gccggcagaa
     2401 gggcacgggc aaggagtacg cagccaagtt catcaagaag cgccgcctgt catccagccg
     2461 gcgtggggtg agccgggagg agatcgagcg ggaggtgaac atcctgcggg agatccggca
     2521 ccccaacatc atcaccctgc acgacatctt cgagaacaag acggacgtgg tcctcatcct
     2581 ggagctggtc tctggcgggg agctctttga cttcctggcg gagaaggagt cgctgacgga
     2641 ggacgaggcc acccagttcc tcaagcagat cctggacggc gttcactacc tgcactctaa
     2701 gcgcatcgca cactttgacc tgaagccgga aaacatcatg ctgctggaca agaacgtgcc
     2761 caacccacga atcaagctca tcgacttcgg catcgcgcac aagatcgagg cggggaacga
     2821 gttcaagaac atcttcggca ccccggagtt tgtggcccca gagattgtga actatgagcc
     2881 gctgggcctg gaggcggaca tgtggagcat cggtgtcatc acctatatcc tcctgagcgg
     2941 tgcatccccg ttcctgggcg agaccaagca ggagacgctc accaacatct cagccgtgaa
     3001 ctacgacttc gacgaggagt acttcagcaa caccagcgag ctggccaagg acttcattcg
     3061 ccggctgctc gtcaaagatc ccaagcggag aatgaccatt gcccagagcc tggaacattc
     3121 ctggattaag gcgatccggc ggcggaacgt gcgtggtgag gacagcggcc gcaagcccga
     3181 gcggcggcgc ctgaagacca cgcgtctgaa ggagtacacc atcaagtcgc actccagctt
     3241 gccgcccaac aacagctacg ccgacttcga gcgcttctcc aaggtgctgg aggaggcggc
     3301 ggccgccgag gagggcctgc gcgagctgca gcgcagccgg cggctctgcc acgaggacgt
     3361 ggaggcgctg gccgccatct acgaggagaa ggaggcctgg taccgcgagg agagcgacag
     3421 cctgggccag gacctgcgga ggctacggca ggagctgctc aagaccgagg cgctcaagcg
     3481 gcaggcgcag gaggaggcca agggcgcgct gctggggacc agcggcctca agcgccgctt
     3541 cagccgcctg gagaaccgct acgaggcgct ggccaagcaa gtagcctccg agatgcgctt
     3601 cgtgcaggac ctcgtgcgcg ccctggagca ggagaagctg cagggcgtgg agtgcgggct
     3661 gcgctaggcg gccgcgaagg atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca
     3721 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacg ggtgcctaga
     3781 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg
     3841 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg
     3901 ggtttgccgc cagaacacag ctgaagcttc gaggggctcg catctctcct tcacgcgccc
     3961 gccgccctac ctgaggccgc catccacgcc ggttgagtcg cgttctgccg cctcccgcct
     4021 gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt aaagctcagg tcgagaccgg
     4081 gcctttgtcc ggcgctccct tggagcctac ctagactcag ccggctctcc acgctttgcc
     4141 tgaccctgct tgctcaactc tacgtctttg tttcgttttc tgttctgcgc cgttacagat
     4201 ccaagctgtg accggcgcct acgctagacg ccaccatgga gagcgacgag agcggcctgc
     4261 ccgccatgga gatcgagtgc cgcatcaccg gcaccctgaa cggcgtggag ttcgagctgg
     4321 tgggcggcgg agagggcacc cccaagcagg gccgcatgac caacaagatg aagagcacca
     4381 aaggcgccct gaccttcagc ccctacctgc tgagccacgt gatgggctac ggcttctacc
     4441 acttcggcac ctaccccagc ggctacgaga accccttcct gcacgccatc aacaacggcg
     4501 gctacaccaa cacccgcatc gagaagtacg aggacggcgg cgtgctgcac gtgagcttca
     4561 gctaccgcta cgaggccggc cgcgtgatcg gcgacttcaa ggtggtgggc accggcttcc
     4621 ccgaggacag cgtgatcttc accgacaaga tcatccgcag caacgccacc gtggagcacc
     4681 tgcaccccat gggcgataac gtgctggtgg gcagcttcgc ccgcaccttc agcctgcgcg
     4741 acggcggcta ctacagcttc gtggtggaca gccacatgca cttcaagagc gccatccacc
     4801 ccagcatcct gcagaacggg ggccccatgt tcgccttccg ccgcgtggag gagctgcaca
     4861 gcaacaccga gctgggcatc gtggagtacc agcacgcctt caagaccccc atcgccttcg
     4921 ccagatcccg cgctcagtcg tccaattctg ccgtggacgg caccgccgga cccggctcca
     4981 ccggatctcg cgagggcaga ggaagtcttc taacatgcgg tgacgtggag gagaatcccg
     5041 gccctatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg
     5101 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc
     5161 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
     5221 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
     5281 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
     5341 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
     5401 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
     5461 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
     5521 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
     5581 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg
     5641 cctgaaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     5701 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     5761 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     5821 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     5881 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     5941 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     6001 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc
     6061 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     6121 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     6181 gtctccgcct tcgccctcag acgagtcgga tctccctttg gccgcctccc cgcctggtac
     6241 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     6301 ggggactgga agggctaatt cactcccaac gaaaataaga tctgcttttt gcttgtactg
     6361 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     6421 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     6481 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     6541 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag
     6601 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     6661 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     6721 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccagttccgc
     6781 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     6841 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag acttttgcag
     6901 agacggccca aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     6961 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     7021 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     7081 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     7141 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     7201 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     7261 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     7321 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     7381 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     7441 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     7501 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     7561 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     7621 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     7681 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     7741 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     7801 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     7861 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7921 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7981 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     8041 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     8101 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     8161 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     8221 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     8281 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     8341 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     8401 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     8461 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     8521 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     8581 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     8641 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     8701 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     8761 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     8821 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     8881 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8941 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     9001 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     9061 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     9121 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     9181 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     9241 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     9301 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     9361 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     9421 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     9481 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     9541 tgccaagctg
//
