LOCUS       V020469                 8850 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020469
VERSION     V020469
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    1798..1915
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2005..2208
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2289..3683
                     /label="PRKN(NM_004562)"
                     /note="PRKN(NM_004562)"
                     /gene="PRKN"
     CDS             3684..3710
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3714..3740
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3744..3770
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     promoter        3820..4031
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4044..4312
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4339..4938
                     /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    4945..5533
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5607..5840
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5912..6033
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6052..6187
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6225..6241)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6249..6265
                     /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(6273..6303)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6318..6339
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6627..7215)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7386..8246)
                     /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(8247..8351)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8825..8841
                     /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 ggttaacttt
     1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga
     1921 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
     2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc
     2281 tagccaccat gatagtgttt gtcaggttca actccagcca tggtttccca gtggaggtcg
     2341 attctgacac cagcatcttc cagctcaagg aggtggttgc taagcgacag ggggttccgg
     2401 ctgaccagtt gcgtgtgatt ttcgcaggga aggagctgag gaatgactgg actgtgcaga
     2461 attgtgacct ggatcagcag agcattgttc acattgtgca gagaccgtgg agaaaaggtc
     2521 aagaaatgaa tgcaactgga ggcgacgacc ccagaaacgc ggcgggaggc tgtgagcggg
     2581 agccccagag cttgactcgg gtggacctca gcagctcagt cctcccagga gactctgtgg
     2641 ggctggctgt cattctgcac actgacagca ggaaggactc accaccagct ggaagtccag
     2701 caggtagatc aatctacaac agcttttatg tgtattgcaa aggcccctgt caaagagtgc
     2761 agccgggaaa actcagggta cagtgcagca cctgcaggca ggcaacgctc accttgaccc
     2821 agggtccatc ttgctgggat gatgttttaa ttccaaaccg gatgagtggt gaatgccaat
     2881 ccccacactg ccctgggact agtgcagaat ttttctttaa atgtggagca caccccacct
     2941 ctgacaagga aacatcagta gctttgcacc tgatcgcaac aaatagtcgg aacatcactt
     3001 gcattacgtg cacagacgtc aggagccccg tcctggtttt ccagtgcaac tcccgccacg
     3061 tgatttgctt agactgtttc cacttatact gtgtgacaag actcaatgat cggcagtttg
     3121 ttcacgaccc tcaacttggc tactccctgc cttgtgtggc tggctgtccc aactccttga
     3181 ttaaagagct ccatcacttc aggattctgg gagaagagca gtacaaccgg taccagcagt
     3241 atggtgcaga ggagtgtgtc ctgcagatgg ggggcgtgtt atgcccccgc cctggctgtg
     3301 gagcggggct gctgccggag cctgaccaga ggaaagtcac ctgcgaaggg ggcaatggcc
     3361 tgggctgtgg gtttgccttc tgccgggaat gtaaagaagc gtaccatgaa ggggagtgca
     3421 gtgccgtatt tgaagcctca ggaacaacta ctcaggccta cagagtcgat gaaagagccg
     3481 ccgagcaggc tcgttgggaa gcagcctcca aagaaaccat caagaaaacc accaagccct
     3541 gtccccgctg ccatgtacca gtggaaaaaa atggaggctg catgcacatg aagtgtccgc
     3601 agccccagtg caggctcgag tggtgctgga actgtggctg cgagtggaac cgcgtctgca
     3661 tgggggacca ctggttcgac gtgtacccat acgacgtacc tgattatgca acttatcctt
     3721 atgatgtacc tgattatgct agctatcctt atgatgtccc agactacgca taggatccgc
     3781 ggccgcgaag gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc acatcgccca
     3841 cagtccccga gaagttgggg ggaggggtcg gcaattgaac gggtgcctag agaaggtggc
     3901 gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg
     3961 gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg
     4021 ccagaacaca gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta
     4081 cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct
     4141 cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg ggcctttgtc
     4201 cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc ctgaccctgc
     4261 ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt
     4321 gaccggcgcc tacgctagat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac
     4381 gacgtccccc gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc
     4441 cacaccgtcg acccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc
     4501 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg
     4561 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc
     4621 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg
     4681 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac
     4741 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc
     4801 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc
     4861 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc
     4921 cgcaagcccg gtgcctgagt cgacaatcaa cctctggatt acaaaatttg tgaaagattg
     4981 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     5041 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     5101 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     5161 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     5221 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     5281 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     5341 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     5401 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     5461 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     5521 gccgcctccc cgcctggtac ctttaagacc aatgacttac aaggcagctg tagatcttag
     5581 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaaaataaga
     5641 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     5701 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     5761 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     5821 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact
     5881 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta
     5941 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     6001 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc
     6061 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     6121 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     6181 ggaggcctag acttttgcag agacggccca aattcgtaat catggtcata gctgtttcct
     6241 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     6301 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     6361 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     6421 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     6481 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     6541 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     6601 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     6661 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     6721 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     6781 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     6841 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6901 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6961 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     7021 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     7081 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     7141 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     7201 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     7261 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     7321 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     7381 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     7441 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     7501 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     7561 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     7621 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     7681 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     7741 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     7801 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     7861 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7921 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7981 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     8041 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     8101 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     8161 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     8221 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     8281 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     8341 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     8401 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     8461 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     8521 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     8581 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
     8641 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc gccattcagg
     8701 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     8761 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     8821 cgttgtaaaa cgacggccag tgccaagctg
//
