LOCUS       V036181                 9272 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036181
VERSION     V036181
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..3109
                     /label="Tigar(NM_177003)"
                     /note="Tigar(NM_177003)"
                     /gene="Tigar"
     CDS             3116..3181
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3225..3436
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3449..3717
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3746..3755
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3776..4438
                     /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             4508..4561
                     /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             4568..5362
                     /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    5369..5957
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6029..6262
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6334..6455
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6474..6609
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6647..6663)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6671..6687
                     /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(6695..6725)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6740..6761
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7049..7637)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7808..8668)
                     /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(8669..8773)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9247..9263
                     /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 ccatgccgcg cttcgccttg accgttatcc gccatggtga
     2341 aacaagactt aataaggaga aaatcattca aggacaaggc gtagatgcgc ccctttcgga
     2401 gactgggttt cggcaagcag cggccgccgg gcagtttctg agcaatgtgc agtttaccca
     2461 cgccttctcc agcgatctca cgaggactaa gcagaccata catggcattt tggagaaaag
     2521 cagattttgt aaagacatgg cggtgaagta cgactccagg ctccgagaaa ggatgtacgg
     2581 agtcgcggaa ggcaagccgc taagcgagct tcgggccatg gccaaagccg ctggggaaga
     2641 gtgccccatg ttcaccccgc ctggaggaga gacagttgag caggtaaaga tgcgcggaaa
     2701 ggatttcttt gatttcattt gtcagctaat cctgggcaag gcagggcaga gagaaagcgt
     2761 cctgcctgga gcgccaggca gcggtttgga aagctctttg gcagaggttt tccctgttgg
     2821 aaaacatggc agcttggggg cgaatcccaa aggtggcacc ctgggcttag cagccagcat
     2881 cttagttgtg agccatggcg cttacatgag aagcctcttt ggttattttc tgagtgacct
     2941 cagatgctcg ttgccaggag cgagagacaa attggaactt tcctccatca ctcccaacac
     3001 tgggatcagt gtcttcatca tagactgtga ggaagcacgc cagccatcga ttcagtgcgt
     3061 ttgtatgaac ctccaggagc acctgaacgg agtgactgag aagcagcacg gatccgacta
     3121 caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa
     3181 gtagcggccg cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc
     3241 gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag
     3301 gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg
     3361 tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt
     3421 tgccgccaga acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg
     3481 ccctacctga ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg
     3541 tgcctcctga actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct
     3601 ttgtccggcg ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac
     3661 cctgcttgct caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa
     3721 gctgtgaccg gcgcctacgc tagacgccac catggagagc gacgagagcg gcctgcccgc
     3781 catggagatc gagtgccgca tcaccggcac cctgaacggc gtggagttcg agctggtggg
     3841 cggcggagag ggcaccccca agcagggccg catgaccaac aagatgaaga gcaccaaagg
     3901 cgccctgacc ttcagcccct acctgctgag ccacgtgatg ggctacggct tctaccactt
     3961 cggcacctac cccagcggct acgagaaccc cttcctgcac gccatcaaca acggcggcta
     4021 caccaacacc cgcatcgaga agtacgagga cggcggcgtg ctgcacgtga gcttcagcta
     4081 ccgctacgag gccggccgcg tgatcggcga cttcaaggtg gtgggcaccg gcttccccga
     4141 ggacagcgtg atcttcaccg acaagatcat ccgcagcaac gccaccgtgg agcacctgca
     4201 ccccatgggc gataacgtgc tggtgggcag cttcgcccgc accttcagcc tgcgcgacgg
     4261 cggctactac agcttcgtgg tggacagcca catgcacttc aagagcgcca tccaccccag
     4321 catcctgcag aacgggggcc ccatgttcgc cttccgccgc gtggaggagc tgcacagcaa
     4381 caccgagctg ggcatcgtgg agtaccagca cgccttcaag acccccatcg ccttcgccag
     4441 atcccgcgct cagtcgtcca attctgccgt ggacggcacc gccggacccg gctccaccgg
     4501 atctcgcgag ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc
     4561 tgccaccatg gctgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     4621 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4681 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4741 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4801 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4861 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4921 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4981 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     5041 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     5101 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     5161 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     5221 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     5281 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     5341 ccttcttgac gagttcttct gagtcgacaa tcaacctctg gattacaaaa tttgtgaaag
     5401 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     5461 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     5521 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     5581 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     5641 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     5701 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     5761 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     5821 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     5881 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     5941 ttgggccgcc tccccgcggt acctttaaga ccaatgactt acaaggcagc tgtagatctt
     6001 agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagataa
     6061 gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     6121 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     6181 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     6241 tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa
     6301 cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt
     6361 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     6421 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc
     6481 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     6541 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     6601 ttggaggcct agacttttgc agagacggcc caaattcgta atcatggtca tagctgtttc
     6661 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     6721 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     6781 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     6841 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6901 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6961 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     7021 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     7081 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     7141 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     7201 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     7261 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     7321 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     7381 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     7441 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     7501 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     7561 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     7621 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     7681 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     7741 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     7801 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     7861 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     7921 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     7981 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     8041 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     8101 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     8161 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     8221 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     8281 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     8341 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     8401 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     8461 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     8521 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     8581 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     8641 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     8701 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     8761 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     8821 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     8881 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     8941 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     9001 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt
     9061 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca
     9121 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     9181 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     9241 gacgttgtaa aacgacggcc agtgccaagc tg
//
