LOCUS       V038272                 9250 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038272
VERSION     V038272
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..1920
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2010..2213
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2302..3357
                     /label="WNT3A(NM_033131)"
                     /note="WNT3A(NM_033131)"
                     /gene="WNT3A"
     promoter        3408..3619
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3632..3900
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3929..3938
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3959..4621
                     /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             4691..4744
                     /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             4745..5344
                     /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    5345..5933
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6007..6240
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6312..6433
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6452..6587
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6625..6641)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6649..6665
                     /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(6673..6703)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6718..6739
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7027..7615)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7786..8646)
                     /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(8647..8751)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9225..9241
                     /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 attacaaaat tcaaaatttt
     1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag
     2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct
     2281 agagctagcg aattcgccac catggcccca ctcggatact tcttactcct ctgcagcctg
     2341 aagcaggctc tgggcagcta cccgatctgg tggtcgctgg ctgttgggcc acagtattcc
     2401 tccctgggct cgcagcccat cctgtgtgcc agcatcccgg gcctggtccc caagcagctc
     2461 cgcttctgca ggaactacgt ggagatcatg cccagcgtgg ccgagggcat caagattggc
     2521 atccaggagt gccagcacca gttccgcggc cgccggtgga actgcaccac cgtccacgac
     2581 agcctggcca tcttcgggcc cgtgctggac aaagctacca gggagtcggc ctttgtccac
     2641 gccattgcct cagccggtgt ggcctttgca gtgacacgct catgtgcaga aggcacggcc
     2701 gccatctgtg gctgcagcag ccgccaccag ggctcaccag gcaagggctg gaagtggggt
     2761 ggctgtagcg aggacatcga gtttggtggg atggtgtctc gggagttcgc cgacgcccgg
     2821 gagaaccggc cagatgcccg ctcagccatg aaccgccaca acaacgaggc tgggcgccag
     2881 gccatcgcca gccacatgca cctcaagtgc aagtgccacg ggctgtcggg cagctgcgag
     2941 gtgaagacat gctggtggtc gcaacccgac ttccgcgcca tcggtgactt cctcaaggac
     3001 aagtacgaca gcgcctcgga gatggtggtg gagaagcacc gggagtcccg cggctgggtg
     3061 gagaccctgc ggccgcgcta cacctacttc aaggtgccca cggagcgcga cctggtctac
     3121 tacgaggcct cgcccaactt ctgcgagccc aaccctgaga cgggctcctt cggcacgcgc
     3181 gaccgcacct gcaacgtcag ctcgcacggc atcgacggct gcgacctgct gtgctgcggc
     3241 cgcggccaca acgcgcgagc ggagcggcgc cgggagaagt gccgctgcgt gttccactgg
     3301 tgctgctacg tcagctgcca ggagtgcacg cgcgtctacg acgtgcacac ctgcaagtag
     3361 ggatccgcgg ccgcgaagga tctgcgatcg ctccggtgcc cgtcagtggg cagagcgcac
     3421 atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaacgg gtgcctagag
     3481 aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga
     3541 gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg
     3601 gtttgccgcc agaacacagc tgaagcttcg aggggctcgc atctctcctt cacgcgcccg
     3661 ccgccctacc tgaggccgcc atccacgccg gttgagtcgc gttctgccgc ctcccgcctg
     3721 tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta aagctcaggt cgagaccggg
     3781 cctttgtccg gcgctccctt ggagcctacc tagactcagc cggctctcca cgctttgcct
     3841 gaccctgctt gctcaactct acgtctttgt ttcgttttct gttctgcgcc gttacagatc
     3901 caagctgtga ccggcgccta cgctagacgc caccatggag agcgacgaga gcggcctgcc
     3961 cgccatggag atcgagtgcc gcatcaccgg caccctgaac ggcgtggagt tcgagctggt
     4021 gggcggcgga gagggcaccc ccaagcaggg ccgcatgacc aacaagatga agagcaccaa
     4081 aggcgccctg accttcagcc cctacctgct gagccacgtg atgggctacg gcttctacca
     4141 cttcggcacc taccccagcg gctacgagaa ccccttcctg cacgccatca acaacggcgg
     4201 ctacaccaac acccgcatcg agaagtacga ggacggcggc gtgctgcacg tgagcttcag
     4261 ctaccgctac gaggccggcc gcgtgatcgg cgacttcaag gtggtgggca ccggcttccc
     4321 cgaggacagc gtgatcttca ccgacaagat catccgcagc aacgccaccg tggagcacct
     4381 gcaccccatg ggcgataacg tgctggtggg cagcttcgcc cgcaccttca gcctgcgcga
     4441 cggcggctac tacagcttcg tggtggacag ccacatgcac ttcaagagcg ccatccaccc
     4501 cagcatcctg cagaacgggg gccccatgtt cgccttccgc cgcgtggagg agctgcacag
     4561 caacaccgag ctgggcatcg tggagtacca gcacgccttc aagaccccca tcgccttcgc
     4621 cagatcccgc gctcagtcgt ccaattctgc cgtggacggc accgccggac ccggctccac
     4681 cggatctcgc gagggcagag gaagtcttct aacatgcggt gacgtggagg agaatcccgg
     4741 ccctatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc
     4801 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc
     4861 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
     4921 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
     4981 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
     5041 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
     5101 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct
     5161 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
     5221 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
     5281 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
     5341 ctgaaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     5401 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     5461 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     5521 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     5581 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     5641 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     5701 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
     5761 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     5821 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     5881 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggtac
     5941 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     6001 ggggactgga agggctaatt cactcccaac gaaaataaga tctgcttttt gcttgtactg
     6061 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     6121 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     6181 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     6241 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag
     6301 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     6361 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     6421 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccagttccgc
     6481 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     6541 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag acttttgcag
     6601 agacggccca aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     6661 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     6721 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     6781 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     6841 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6901 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6961 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     7021 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     7081 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     7141 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     7201 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     7261 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     7321 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     7381 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     7441 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     7501 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     7561 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7621 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7681 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7741 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7801 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7861 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7921 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7981 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     8041 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     8101 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     8161 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     8221 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     8281 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     8341 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     8401 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     8461 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     8521 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     8581 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8641 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8701 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8761 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     8821 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     8881 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     8941 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     9001 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     9061 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     9121 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     9181 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     9241 tgccaagctg
//
