LOCUS       V028764                 8884 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028764
VERSION     V028764
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..2910
                     /label="Denr(NM_026603)"
                     /note="Denr(NM_026603)"
                     /gene="Denr"
     CDS             2917..2982
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3026..3237
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3250..3518
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3547..3556
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3577..4239
                     /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             4309..4362
                     /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             4363..4962
                     /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    4963..5551
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5625..5858
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5930..6051
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6091..6226
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6259..6275)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6283..6299
                     /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(6307..6337)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6352..6373
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6661..7249)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7420..8280)
                     /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(8281..8385)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8859..8875
                     /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 ctactgacat ttctgaatcc
     2341 agcggggcgg attgcaaagg cgatacgaag aacagtgcca agttagatgc ggattaccct
     2401 cttcgagtcc tttattgtgg agtctgttca ttaccaacag agtactgtga atacatgccg
     2461 gatgttgcta aatgtagaca atggctagag aagaactttc caaatgaatt tgcaaaactt
     2521 actgtagaaa attcacccaa acaagaaact ggaattactg aaggccaagg accagtaggg
     2581 gaagaggagg agaagaaaaa acaaaaaaga ggtggaagag gtcaaataaa acagaagaag
     2641 aagacagtac cacagaaggt cacgatagcc aaaattccca gagcaaagaa gaaatacgtg
     2701 acgagagtgt gtggccttgc aacttttgaa attgatctca aagaagcaca aagatttttt
     2761 gctcaaaaat tctcgtgcgg tgcctcagtg acaggggagg acgagatcat cattcaggga
     2821 gacttcacag atgacatcat tgacgtcatt caggaaaagt ggccagaggt ggatgacgac
     2881 agcattgaag accttggaga agtgaagaag ggatccgact acaaagacca tgacggtgat
     2941 tataaagatc atgacatcga ctacaaggat gacgatgaca agtagcggcc gcgaaggatc
     3001 tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag
     3061 ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg
     3121 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata
     3181 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagctg
     3241 aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat
     3301 ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc
     3361 gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg
     3421 agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctac
     3481 gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc ggcgcctacg
     3541 ctagacgcca ccatggagag cgacgagagc ggcctgcccg ccatggagat cgagtgccgc
     3601 atcaccggca ccctgaacgg cgtggagttc gagctggtgg gcggcggaga gggcaccccc
     3661 aagcagggcc gcatgaccaa caagatgaag agcaccaaag gcgccctgac cttcagcccc
     3721 tacctgctga gccacgtgat gggctacggc ttctaccact tcggcaccta ccccagcggc
     3781 tacgagaacc ccttcctgca cgccatcaac aacggcggct acaccaacac ccgcatcgag
     3841 aagtacgagg acggcggcgt gctgcacgtg agcttcagct accgctacga ggccggccgc
     3901 gtgatcggcg acttcaaggt ggtgggcacc ggcttccccg aggacagcgt gatcttcacc
     3961 gacaagatca tccgcagcaa cgccaccgtg gagcacctgc accccatggg cgataacgtg
     4021 ctggtgggca gcttcgcccg caccttcagc ctgcgcgacg gcggctacta cagcttcgtg
     4081 gtggacagcc acatgcactt caagagcgcc atccacccca gcatcctgca gaacgggggc
     4141 cccatgttcg ccttccgccg cgtggaggag ctgcacagca acaccgagct gggcatcgtg
     4201 gagtaccagc acgccttcaa gacccccatc gccttcgcca gatcccgcgc tcagtcgtcc
     4261 aattctgccg tggacggcac cgccggaccc ggctccaccg gatctcgcga gggcagagga
     4321 agtcttctaa catgcggtga cgtggaggag aatcccggcc ctatgaccga gtacaagccc
     4381 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     4441 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     4501 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     4561 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     4621 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     4681 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     4741 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     4801 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     4861 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     4921 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaaatcaacc tctggattac
     4981 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     5041 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     5101 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     5161 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     5221 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     5281 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     5341 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg
     5401 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     5461 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     5521 agtcggatct ccctttgggc cgcctccccg cctggtacct ttaagaccaa tgacttacaa
     5581 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca
     5641 ctcccaacga agataagatc tgctttttgc ttgtactggg tctctctggt tagaccagat
     5701 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     5761 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     5821 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat
     5881 tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat
     5941 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     6001 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     6061 gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     6121 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     6181 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctaga cttttgcaga
     6241 gaccaaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     6301 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     6361 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     6421 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     6481 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     6541 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     6601 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     6661 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6721 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6781 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6841 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6901 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6961 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     7021 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     7081 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     7141 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     7201 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     7261 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     7321 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     7381 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     7441 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     7501 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     7561 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     7621 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     7681 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     7741 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     7801 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     7861 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     7921 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     7981 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     8041 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     8101 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     8161 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     8221 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     8281 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     8341 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     8401 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     8461 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     8521 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     8581 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     8641 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     8701 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     8761 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     8821 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa
     8881 gctg
//
