LOCUS       V024026                 9017 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024026
VERSION     V024026
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..3060
                     /label="Ifi30(NM_023065)"
                     /note="Ifi30(NM_023065)"
                     /gene="Ifi30"
     CDS             3067..3132
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3176..3387
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3400..3668
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3697..3706
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3727..4389
                     /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             4459..4512
                     /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             4513..5112
                     /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    5113..5700
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5774..6007
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6079..6200
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6219..6354
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6392..6408)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6416..6432
                     /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(6440..6470)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6485..6506
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6794..7382)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7553..8413)
                     /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(8414..8518)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8992..9008
                     /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 gccaccatgt cctggtctcc gatcctacca
     2341 tttttgtccc ttctgcttct gctgttccca ctcgaggttc ccagagcagc cactgcgtcc
     2401 ctttcgcaag catcctccga aggcacaacc acctgcaagg cccacgatgt gtgcctcttg
     2461 gggccacgcc cattgccccc ttcaccacct gtcagagtca gcctgtatta tgagtccctg
     2521 tgtggagctt gtcgctactt cctcgtccgg gatttgttcc caacctggct gatggtgatg
     2581 gaaatcatga acatcaccct ggtgccctac gggaatgcac aggagagaaa tgtcagcggt
     2641 acgtgggagt tcacgtgcca gcacggggag ctggagtgta gactgaacat ggtggaggcc
     2701 tgtctgctgg ataagctgga aaaggaggca gcgttcctaa ccatcgtctg tatggaggag
     2761 atggatgata tggagaagaa actgggaccg tgcctgcagg tgtatgctcc tgaggtgtca
     2821 ccagagagta tcatggagtg cgccacagga aaacggggca cacagctcat gcatgagaat
     2881 gcccagctca cagatgccct acacccaccc cacgagtatg tgccctgggt gctggtcaat
     2941 gagaaacctt tgaaggaccc cagcgagctc ctgagcatag tctgtcagct gtaccaggga
     3001 acggagaagc cagacatctg ctcctccatt gccgactccc ccaggaaggt ctgctataag
     3061 ggatccgact acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat
     3121 gacgatgaca agtagcggcc gcgaaggatc tgcgatcgct ccggtgcccg tcagtgggca
     3181 gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaacgggt
     3241 gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt
     3301 tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt
     3361 cgcaacgggt ttgccgccag aacacagctg aagcttcgag gggctcgcat ctctccttca
     3421 cgcgcccgcc gccctacctg aggccgccat ccacgccggt tgagtcgcgt tctgccgcct
     3481 cccgcctgtg gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg
     3541 agaccgggcc tttgtccggc gctcccttgg agcctaccta gactcagccg gctctccacg
     3601 ctttgcctga ccctgcttgc tcaactctac gtctttgttt cgttttctgt tctgcgccgt
     3661 tacagatcca agctgtgacc ggcgcctacg ctagacgcca ccatggagag cgacgagagc
     3721 ggcctgcccg ccatggagat cgagtgccgc atcaccggca ccctgaacgg cgtggagttc
     3781 gagctggtgg gcggcggaga gggcaccccc aagcagggcc gcatgaccaa caagatgaag
     3841 agcaccaaag gcgccctgac cttcagcccc tacctgctga gccacgtgat gggctacggc
     3901 ttctaccact tcggcaccta ccccagcggc tacgagaacc ccttcctgca cgccatcaac
     3961 aacggcggct acaccaacac ccgcatcgag aagtacgagg acggcggcgt gctgcacgtg
     4021 agcttcagct accgctacga ggccggccgc gtgatcggcg acttcaaggt ggtgggcacc
     4081 ggcttccccg aggacagcgt gatcttcacc gacaagatca tccgcagcaa cgccaccgtg
     4141 gagcacctgc accccatggg cgataacgtg ctggtgggca gcttcgcccg caccttcagc
     4201 ctgcgcgacg gcggctacta cagcttcgtg gtggacagcc acatgcactt caagagcgcc
     4261 atccacccca gcatcctgca gaacgggggc cccatgttcg ccttccgccg cgtggaggag
     4321 ctgcacagca acaccgagct gggcatcgtg gagtaccagc acgccttcaa gacccccatc
     4381 gccttcgcca gatcccgcgc tcagtcgtcc aattctgccg tggacggcac cgccggaccc
     4441 ggctccaccg gatctcgcga gggcagagga agtcttctaa catgcggtga cgtggaggag
     4501 aatcccggcc ctatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     4561 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     4621 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     4681 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     4741 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     4801 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     4861 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     4921 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     4981 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     5041 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     5101 cccggtgcct gaaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     5161 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     5221 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     5281 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     5341 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     5401 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     5461 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt
     5521 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     5581 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     5641 cttccgcgtc tccgccttcg ccctcagacg agtcggatct ccctttggcc gcctccccgc
     5701 ctggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     5761 gaaaaggggg gactggaagg gctaattcac tcccaacgaa aataagatct gctttttgct
     5821 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     5881 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     5941 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     6001 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga
     6061 atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     6121 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     6181 aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca
     6241 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     6301 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact
     6361 tttgcagaga cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     6421 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     6481 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6541 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6601 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6661 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6721 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6781 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6841 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6901 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6961 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     7021 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     7081 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     7141 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7201 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     7261 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7321 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7381 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7441 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7501 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7561 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7621 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7681 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7741 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7801 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7861 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7921 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7981 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     8041 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     8101 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     8161 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     8221 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     8281 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     8341 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     8401 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8461 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8521 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     8581 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     8641 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     8701 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     8761 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     8821 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt
     8881 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     8941 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     9001 cggccagtgc caagctg
//
