LOCUS       V034589                 9420 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034589
VERSION     V034589
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             2317..3615
                     /label="PRMT2(NM_206962)"
                     /note="PRMT2(NM_206962)"
                     /gene="PRMT2"
     CDS             3622..3648
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3652..3678
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3682..3708
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     promoter        3773..3984
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3997..4265
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4294..4303
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4324..4986
                     /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             5056..5109
                     /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             5110..5508
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /note="confers resistance to blasticidin"
                     /product="blasticidin S deaminase"
                     /transl_table=1
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG*"
                     /label="BSD"
     misc_feature    5515..6103
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6177..6410
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6482..6603
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6622..6757
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6795..6811)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6819..6835
                     /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(6843..6873)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6888..6909
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7197..7785)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7956..8816)
                     /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(8817..8921)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9395..9411
                     /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 ctgatacgaa ctcggaattc gccaccatgg caacatcagg tgactgtccc
     2341 agaagtgaat cgcagggaga agagcctgct gagtgcagtg aggccggtct cctgcaggag
     2401 ggagtacagc cagaggagtt tgtggccatc gcggactacg ctgccaccga tgagacccag
     2461 ctcagttttt tgagaggaga aaaaattctt atcctgagac aaaccactgc agattggtgg
     2521 tggggtgagc gtgcgggctg ctgtgggtac attccggcaa accatgtggg gaagcacgtg
     2581 gatgagtacg accccgagga cacgtggcag gatgaagagt acttcggcag ctatggaact
     2641 ctgaaactcc acttggagat gttggcagac cagccacgaa caactaaata ccacagtgtc
     2701 atcctgcaga ataaagaatc cctgacggat aaagtcatcc tggacgtggg ctgtgggact
     2761 gggatcatca gtctcttctg tgcacactat gcgcggccta gagcggtgta cgcggtggag
     2821 gccagtgaga tggcacagca cacggggcag ctggtcctgc agaacggctt tgctgacatc
     2881 atcaccgtgt accagcagaa ggtggaggat gtggtgctgc ccgagaaggt ggacgtgctg
     2941 gtgtctgagt ggatggggac ctgcctgctg tttgagttca tgatcgagtc catcctgtat
     3001 gcccgggatg cctggctgaa ggaggacggg gtcatttggc ccaccatggc tgcgttgcac
     3061 cttgtgccct gcagtgctga taaggattat cgtagcaagg tgctcttctg ggacaacgcg
     3121 tacgagttca acctcagcgc tctgaaatct ttagcagtta aggagttttt ttcaaagccc
     3181 aagtataacc acattttgaa accagaagac tgtctctctg aaccgtgcac tatattgcag
     3241 ttggacatga gaaccgtgca aatttctgat ctagagaccc tgaggggcga gctgcgcttc
     3301 gacatcagga aggcggggac cctgcacggc ttcacggcct ggtttagcgt ccacttccag
     3361 agcctgcagg aggggcagcc gccgcaggtg ctcagcaccg ggcccttcca ccccaccaca
     3421 cactggaagc agacgctgtt catgatggac gacccagtcc ctgtccatac aggagacgtg
     3481 gtcacgggtt cagttgtgtt gcagagaaac ccagtgtgga gaaggcacat gtctgtggct
     3541 ctgagctggg ctgtcacttc cagacaagac cccacatctc aaaaagttgg agaaaaagtc
     3601 ttccccatct ggagactcga gtacccatac gacgtacctg attatgcaac ttatccttat
     3661 gatgtacctg attatgctag ctatccttat gatgtcccag actacgcata gggatccctg
     3721 cgatatccag cgcggccgcg aaggatctgc gatcgctccg gtgcccgtca gtgggcagag
     3781 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aacgggtgcc
     3841 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt
     3901 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc
     3961 aacgggtttg ccgccagaac acagctgaag cttcgagggg ctcgcatctc tccttcacgc
     4021 gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc
     4081 gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga
     4141 ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt
     4201 tgcctgaccc tgcttgctca actctacgtc tttgtttcgt tttctgttct gcgccgttac
     4261 agatccaagc tgtgaccggc gcctacgcta gacgccacca tggagagcga cgagagcggc
     4321 ctgcccgcca tggagatcga gtgccgcatc accggcaccc tgaacggcgt ggagttcgag
     4381 ctggtgggcg gcggagaggg cacccccaag cagggccgca tgaccaacaa gatgaagagc
     4441 accaaaggcg ccctgacctt cagcccctac ctgctgagcc acgtgatggg ctacggcttc
     4501 taccacttcg gcacctaccc cagcggctac gagaacccct tcctgcacgc catcaacaac
     4561 ggcggctaca ccaacacccg catcgagaag tacgaggacg gcggcgtgct gcacgtgagc
     4621 ttcagctacc gctacgaggc cggccgcgtg atcggcgact tcaaggtggt gggcaccggc
     4681 ttccccgagg acagcgtgat cttcaccgac aagatcatcc gcagcaacgc caccgtggag
     4741 cacctgcacc ccatgggcga taacgtgctg gtgggcagct tcgcccgcac cttcagcctg
     4801 cgcgacggcg gctactacag cttcgtggtg gacagccaca tgcacttcaa gagcgccatc
     4861 caccccagca tcctgcagaa cgggggcccc atgttcgcct tccgccgcgt ggaggagctg
     4921 cacagcaaca ccgagctggg catcgtggag taccagcacg ccttcaagac ccccatcgcc
     4981 ttcgccagat cccgcgctca gtcgtccaat tctgccgtgg acggcaccgc cggacccggc
     5041 tccaccggat ctcgcgaggg cagaggaagt cttctaacat gcggtgacgt ggaggagaat
     5101 cccggcccta tggccaagcc tttgtctcaa gaagaatcca ccctcattga aagagcaacg
     5161 gctacaatca acagcatccc catctctgaa gactacagcg tcgccagcgc agctctctct
     5221 agcgacggcc gcatcttcac tggtgtcaat gtatatcatt ttactggggg accttgtgca
     5281 gaactcgtgg tgctgggcac tgctgctgct gcggcagctg gcaacctgac ttgtatcgtc
     5341 gcgatcggaa atgagaacag gggcatcttg agcccctgcg gacggtgccg acaggtgctt
     5401 ctcgatctgc atcctgggat caaagccata gtgaaggaca gtgatggaca gccgacggca
     5461 gttgggattc gtgaattgct gccctctggt tatgtgtggg agggctaagt cgacaatcaa
     5521 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     5581 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     5641 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     5701 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     5761 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     5821 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     5881 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     5941 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     6001 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     6061 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggtac ctttaagacc
     6121 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga
     6181 agggctaatt cactcccaac gaaaataaga tctgcttttt gcttgtactg ggtctctctg
     6241 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     6301 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     6361 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc
     6421 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag
     6481 gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     6541 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     6601 ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccagttccgc ccattctccg
     6661 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     6721 ctattccaga agtagtgagg aggctttttt ggaggcctag acttttgcag agacggccca
     6781 aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     6841 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     6901 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     6961 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     7021 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     7081 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     7141 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     7201 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     7261 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     7321 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     7381 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     7441 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     7501 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     7561 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     7621 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     7681 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     7741 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     7801 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     7861 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     7921 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     7981 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     8041 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     8101 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     8161 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     8221 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     8281 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     8341 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     8401 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     8461 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     8521 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     8581 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     8641 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     8701 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     8761 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     8821 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     8881 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     8941 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     9001 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     9061 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     9121 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     9181 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     9241 taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     9301 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     9361 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctg
//
