LOCUS       V021833                 9164 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021833
VERSION     V021833
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..3043
                     /label="Gzmb(NM_013542)"
                     /note="Gzmb(NM_013542)"
                     /gene="Gzmb"
     CDS             3044..3073
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     promoter        3117..3328
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3341..3609
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3638..3647
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3668..4330
                     /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             4400..4453
                     /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             4460..5254
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     misc_feature    5261..5849
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5921..6154
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6226..6347
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6366..6501
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6539..6555)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6563..6579
                     /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(6587..6617)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6632..6653
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6941..7529)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7700..8560)
                     /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(8561..8665)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9139..9155
                     /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 ccatgaagat cctcctgcta ctgctgacct tgtctctggc
     2341 ctccaggaca aaggcagggg agatcatcgg gggacatgaa gtcaagcccc actctcgacc
     2401 ctacatggcc ttactttcga tcaaggatca gcagcctgag gcgatatgtg ggggcttcct
     2461 tattcgagag gactttgtgc tgactgctgc tcactgtgaa ggaagtataa taaatgtcac
     2521 tttgggggcc cacaacatca aagaacagga gaagacccag caagtcatcc ctatggtaaa
     2581 atgcattccc cacccagact ataatcctaa gacattctcc aatgacatca tgctgctaaa
     2641 gctgaagagt aaggccaaga ggactagagc tgtgaggccc ctcaacctgc ccaggcgcaa
     2701 tgtcaatgtg aagccaggag atgtgtgcta tgtggctggt tggggaagga tggccccaat
     2761 gggcaaatac tcaaacacgc tacaagaggt tgagctgaca gtacagaagg atcgggagtg
     2821 tgagtcctac tttaaaaatc gttacaacaa aaccaatcag atatgtgcgg gggacccaaa
     2881 gaccaaacgt gcttcctttc ggggggattc tggaggcccg cttgtgtgta aaaaagtggc
     2941 tgcaggcata gtttcctatg gatataagga tggttcacct ccacgtgctt tcaccaaagt
     3001 ctcgagtttc ttatcctgga taaagaaaac aatgaaaagc agcgagcaaa agctcatttc
     3061 tgaagaggac ttgtagcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc
     3121 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg
     3181 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
     3241 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
     3301 tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc
     3361 acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc
     3421 tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc
     3481 gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac
     3541 gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg
     3601 ttacagatcc aagctgtgac cggcgcctac gctagacgcc accatggaga gcgacgagag
     3661 cggcctgccc gccatggaga tcgagtgccg catcaccggc accctgaacg gcgtggagtt
     3721 cgagctggtg ggcggcggag agggcacccc caagcagggc cgcatgacca acaagatgaa
     3781 gagcaccaaa ggcgccctga ccttcagccc ctacctgctg agccacgtga tgggctacgg
     3841 cttctaccac ttcggcacct accccagcgg ctacgagaac cccttcctgc acgccatcaa
     3901 caacggcggc tacaccaaca cccgcatcga gaagtacgag gacggcggcg tgctgcacgt
     3961 gagcttcagc taccgctacg aggccggccg cgtgatcggc gacttcaagg tggtgggcac
     4021 cggcttcccc gaggacagcg tgatcttcac cgacaagatc atccgcagca acgccaccgt
     4081 ggagcacctg caccccatgg gcgataacgt gctggtgggc agcttcgccc gcaccttcag
     4141 cctgcgcgac ggcggctact acagcttcgt ggtggacagc cacatgcact tcaagagcgc
     4201 catccacccc agcatcctgc agaacggggg ccccatgttc gccttccgcc gcgtggagga
     4261 gctgcacagc aacaccgagc tgggcatcgt ggagtaccag cacgccttca agacccccat
     4321 cgccttcgcc agatcccgcg ctcagtcgtc caattctgcc gtggacggca ccgccggacc
     4381 cggctccacc ggatctcgcg agggcagagg aagtcttcta acatgcggtg acgtggagga
     4441 gaatcccggc cctgccacca tggctgaaca agatggattg cacgcaggtt ctccggccgc
     4501 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     4561 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     4621 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4681 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4741 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     4801 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     4861 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     4921 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4981 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     5041 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     5101 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     5161 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     5221 cgccttctat cgccttcttg acgagttctt ctgagtcgac aatcaacctc tggattacaa
     5281 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     5341 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     5401 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     5461 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     5521 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     5581 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     5641 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
     5701 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     5761 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     5821 tcggatctcc ctttgggccg cctccccgcg gtacctttaa gaccaatgac ttacaaggca
     5881 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc
     5941 caacgaaaat aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga
     6001 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     6061 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     6121 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt
     6181 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca
     6241 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     6301 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc
     6361 tagctatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     6421 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     6481 gaggaggctt ttttggaggc ctagactttt gcagagacgg cccaaattcg taatcatggt
     6541 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     6601 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     6661 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     6721 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     6781 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     6841 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6901 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6961 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     7021 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     7081 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     7141 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     7201 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     7261 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     7321 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     7381 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     7441 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     7501 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     7561 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     7621 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     7681 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     7741 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     7801 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     7861 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     7921 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     7981 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     8041 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     8101 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     8161 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     8221 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     8281 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     8341 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     8401 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     8461 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     8521 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     8581 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     8641 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     8701 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     8761 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     8821 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     8881 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     8941 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
     9001 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     9061 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     9121 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctg
//
