LOCUS       V024065                 9218 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024065
VERSION     V024065
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..3261
                     /label="Calu(NM_007594)"
                     /note="Calu(NM_007594)"
                     /gene="Calu"
     CDS             3268..3333
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3377..3588
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3601..3869
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3898..3907
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3928..4590
                     /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             4660..4713
                     /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             4714..5313
                     /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    5314..5901
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5975..6208
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6280..6401
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6420..6555
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6593..6609)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6617..6633
                     /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(6641..6671)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6686..6707
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6995..7583)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7754..8614)
                     /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(8615..8719)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9193..9209
                     /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 acctgcgtca gtttcttatg
     2341 tgcctgtccc tgtgcacagc ctttgctttg agcaagccta cagaaaagaa ggaccgagta
     2401 caccatgagc ctcagctcag cgataaagtt cacaatgatg ctcagaattt tgactatgac
     2461 catgatgcct tcttgggtgc agaagaggca aagagttttg atcagctgac accagaagag
     2521 agcaaggaaa ggcttggaaa gattgtaagt aaaatagatg acgacaagga tgggtttgtc
     2581 actgtggatg aactcaaagg ctggattaag tttgcacaaa agcgctggat tcacgaggat
     2641 gtagagcggc aatggaaggg gcacgacctc aatgaggatg gcctcgtttc ctgggaggag
     2701 tataaaaatg ccacctacgg ctacgtttta gatgatccag atcctgatga tggatttaat
     2761 tataaacaga tgatggtcag agatgagcgg aggtttaaaa tggcagacaa ggatggagac
     2821 ctaattgcca caaaggaaga gttcacagct ttcctgcacc ctgaggaata tgactacatg
     2881 aaagacatag tcgtgcagga aaccatggag gatatagaca agaatgctga tgggttcatt
     2941 gatctagagg agtacattgg tgacatgtac agtcatgatg ggaatgctga tgagccagag
     3001 tgggtgaaga cagagcgaga acagttcgtg gagtttcgag ataagaaccg ggatgggaag
     3061 atggacaagg aagagaccaa agactggatc ctcccctcag actatgacca tgcagaggca
     3121 gaagccaggc atctggtcta tgagtcagac caaaacaagg atggcaagct caccaaggag
     3181 gagattgtcg acaagtatga tttatttgtg ggcagccagg ccacagattt cggggaggcc
     3241 ttagtacgac atgatgagtt cggatccgac tacaaagacc atgacggtga ttataaagat
     3301 catgacatcg actacaagga tgacgatgac aagtagcggc cgcgaaggat ctgcgatcgc
     3361 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga
     3421 ggggtcggca attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat
     3481 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta
     3541 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga
     3601 ggggctcgca tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg
     3661 ttgagtcgcg ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag
     3721 gtaagtttaa agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct
     3781 agactcagcc ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt
     3841 tcgttttctg ttctgcgccg ttacagatcc aagctgtgac cggcgcctac gctagacgcc
     3901 accatggaga gcgacgagag cggcctgccc gccatggaga tcgagtgccg catcaccggc
     3961 accctgaacg gcgtggagtt cgagctggtg ggcggcggag agggcacccc caagcagggc
     4021 cgcatgacca acaagatgaa gagcaccaaa ggcgccctga ccttcagccc ctacctgctg
     4081 agccacgtga tgggctacgg cttctaccac ttcggcacct accccagcgg ctacgagaac
     4141 cccttcctgc acgccatcaa caacggcggc tacaccaaca cccgcatcga gaagtacgag
     4201 gacggcggcg tgctgcacgt gagcttcagc taccgctacg aggccggccg cgtgatcggc
     4261 gacttcaagg tggtgggcac cggcttcccc gaggacagcg tgatcttcac cgacaagatc
     4321 atccgcagca acgccaccgt ggagcacctg caccccatgg gcgataacgt gctggtgggc
     4381 agcttcgccc gcaccttcag cctgcgcgac ggcggctact acagcttcgt ggtggacagc
     4441 cacatgcact tcaagagcgc catccacccc agcatcctgc agaacggggg ccccatgttc
     4501 gccttccgcc gcgtggagga gctgcacagc aacaccgagc tgggcatcgt ggagtaccag
     4561 cacgccttca agacccccat cgccttcgcc agatcccgcg ctcagtcgtc caattctgcc
     4621 gtggacggca ccgccggacc cggctccacc ggatctcgcg agggcagagg aagtcttcta
     4681 acatgcggtg acgtggagga gaatcccggc cctatgaccg agtacaagcc cacggtgcgc
     4741 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac
     4801 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg
     4861 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     4921 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     4981 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     5041 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     5101 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     5161 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     5221 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     5281 acctggtgca tgacccgcaa gcccggtgcc tgaaatcaac ctctggatta caaaatttgt
     5341 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     5401 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     5461 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     5521 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     5581 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     5641 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     5701 tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc
     5761 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     5821 ctgctgccgg ctctgcggcc tcttccgcgt ctccgccttc gccctcagac gagtcggatc
     5881 tccctttggc cgcctccccg cctggtacct ttaagaccaa tgacttacaa ggcagctgta
     5941 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
     6001 aaataagatc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg
     6061 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     6121 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     6181 ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat
     6241 ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat
     6301 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     6361 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta
     6421 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     6481 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     6541 gcttttttgg aggcctagac ttttgcagag acggcccaaa ttcgtaatca tggtcatagc
     6601 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     6661 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     6721 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     6781 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     6841 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     6901 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     6961 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     7021 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     7081 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     7141 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     7201 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     7261 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     7321 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     7381 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     7441 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     7501 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     7561 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     7621 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     7681 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     7741 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     7801 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     7861 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     7921 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     7981 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     8041 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     8101 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     8161 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     8221 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     8281 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     8341 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     8401 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     8461 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     8521 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     8581 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     8641 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     8701 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     8761 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     8821 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     8881 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     8941 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     9001 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
     9061 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     9121 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     9181 agtcacgacg ttgtaaaacg acggccagtg ccaagctg
//
