LOCUS       V020502                 9439 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020502
VERSION     V020502
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             2297..3547
                     /label="SIRT3(NM_001370310)"
                     /note="SIRT3(NM_001370310)"
                     /gene="SIRT3"
     promoter        3598..3809
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3822..4090
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4119..4128
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4149..4811
                     /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             4881..4934
                     /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             4935..5534
                     /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    5535..6122
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6196..6429
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6501..6622
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6641..6776
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6814..6830)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6838..6854
                     /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(6862..6892)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6907..6928
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7216..7804)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7975..8835)
                     /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(8836..8940)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9414..9430
                     /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 gccaccatgg cgttctgggg ttggcgcgcc gcggcagccc tccggctgtg
     2341 gggccgggta gttgaacggg tcgaggccgg gggaggcgtg gggccgtttc aggcctgcgg
     2401 ctgtcggctg gtgcttggcg gcagggacga tgtgagtgcg gggctgagag gcagccatgg
     2461 ggcccgcggt gagcccttgg acccggcgcg ccccttgcag aggcctccca gacccgaggt
     2521 gcccagggca ttccggaggc agccgagggc agcagctccc agtttcttct tttcgagtat
     2581 taaaggtgga agaaggtcca tatctttttc tgtgggtgct tcaagtgttg ttggaagtgg
     2641 aggcagcagt gacaagggga agctttccct gcaggatgta gctgagctga ttcgggccag
     2701 agcctgccag agggtggtgg tcatggtggg ggccggcatc agcacaccca gtggcattcc
     2761 agacttcaga tcgccgggga gtggcctgta cagcaacctc cagcagtacg atctcccgta
     2821 ccccgaggcc atttttgaac tcccattctt ctttcacaac cccaagccct ttttcacttt
     2881 ggccaaggag ctgtaccctg gaaactacaa gcccaacgtc actcactact ttctccggct
     2941 gcttcatgac aaggggctgc ttctgcggct ctacacgcag aacatcgatg ggcttgagag
     3001 agtgtcgggc atccctgcct caaagctggt tgaagctcat ggaacctttg cctctgccac
     3061 ctgcacagtc tgccaaagac ccttcccagg ggaggacatt cgggctgacg tgatggcaga
     3121 cagggttccc cgctgcccgg tctgcaccgg cgttgtgaag cccgacattg tgttctttgg
     3181 ggagccgctg ccccagaggt tcttgctgca tgtggttgat ttccccatgg cagatctgct
     3241 gctcatcctt gggacctccc tggaggtgga gccttttgcc agcttgaccg aggccgtgcg
     3301 gagctcagtt ccccgactgc tcatcaaccg ggacttggtg gggcccttgg cttggcatcc
     3361 tcgcagcagg gacgtggccc agctggggga cgtggttcac ggcgtggaaa gcctagtgga
     3421 gcttctgggc tggacagaag agatgcggga ccttgtgcag cgggaaactg ggaaggtaca
     3481 gactgctgag gaggaccatc caaggggctg cccttctcac tgctctaggc ttgatggacc
     3541 agacaaatag ggatccgcgg ccgcgaagga tctgcgatcg ctccggtgcc cgtcagtggg
     3601 cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaacgg
     3661 gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc
     3721 tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt
     3781 ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc atctctcctt
     3841 cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc gttctgccgc
     3901 ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta aagctcaggt
     3961 cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc cggctctcca
     4021 cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct gttctgcgcc
     4081 gttacagatc caagctgtga ccggcgccta cgctagacgc caccatggag agcgacgaga
     4141 gcggcctgcc cgccatggag atcgagtgcc gcatcaccgg caccctgaac ggcgtggagt
     4201 tcgagctggt gggcggcgga gagggcaccc ccaagcaggg ccgcatgacc aacaagatga
     4261 agagcaccaa aggcgccctg accttcagcc cctacctgct gagccacgtg atgggctacg
     4321 gcttctacca cttcggcacc taccccagcg gctacgagaa ccccttcctg cacgccatca
     4381 acaacggcgg ctacaccaac acccgcatcg agaagtacga ggacggcggc gtgctgcacg
     4441 tgagcttcag ctaccgctac gaggccggcc gcgtgatcgg cgacttcaag gtggtgggca
     4501 ccggcttccc cgaggacagc gtgatcttca ccgacaagat catccgcagc aacgccaccg
     4561 tggagcacct gcaccccatg ggcgataacg tgctggtggg cagcttcgcc cgcaccttca
     4621 gcctgcgcga cggcggctac tacagcttcg tggtggacag ccacatgcac ttcaagagcg
     4681 ccatccaccc cagcatcctg cagaacgggg gccccatgtt cgccttccgc cgcgtggagg
     4741 agctgcacag caacaccgag ctgggcatcg tggagtacca gcacgccttc aagaccccca
     4801 tcgccttcgc cagatcccgc gctcagtcgt ccaattctgc cgtggacggc accgccggac
     4861 ccggctccac cggatctcgc gagggcagag gaagtcttct aacatgcggt gacgtggagg
     4921 agaatcccgg ccctatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg
     4981 tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca
     5041 ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc
     5101 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct
     5161 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg
     5221 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc
     5281 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg
     5341 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg
     5401 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct
     5461 tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca
     5521 agcccggtgc ctgaaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     5581 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     5641 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     5701 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     5761 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     5821 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     5881 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct
     5941 ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     6001 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     6061 ctcttccgcg tctccgcctt cgccctcaga cgagtcggat ctccctttgg ccgcctcccc
     6121 gcctggtacc tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
     6181 aagaaaaggg gggactggaa gggctaattc actcccaacg aaaataagat ctgctttttg
     6241 cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
     6301 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
     6361 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
     6421 tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat
     6481 gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca
     6541 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     6601 ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc
     6661 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     6721 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctaga
     6781 cttttgcaga gacggcccaa attcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     6841 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     6901 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     6961 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     7021 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     7081 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     7141 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     7201 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     7261 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     7321 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     7381 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     7441 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     7501 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     7561 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     7621 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     7681 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     7741 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     7801 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     7861 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     7921 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     7981 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     8041 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     8101 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     8161 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     8221 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     8281 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     8341 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     8401 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     8461 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     8521 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     8581 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     8641 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     8701 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     8761 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     8821 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     8881 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     8941 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     9001 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     9061 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     9121 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     9181 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     9241 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
     9301 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     9361 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     9421 gacggccagt gccaagctg
//
