LOCUS       V024718                 8903 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024718
VERSION     V024718
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..2946
                     /label="Isoc2b(NM_026158)"
                     /note="Isoc2b(NM_026158)"
                     /gene="Isoc2b"
     CDS             2953..3018
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3062..3273
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3286..3554
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3583..3592
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3613..4275
                     /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             4345..4398
                     /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             4399..4998
                     /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    4999..5586
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5660..5893
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5965..6086
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6105..6240
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6278..6294)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6302..6318
                     /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(6326..6356)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6371..6392
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6680..7268)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7439..8299)
                     /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(8300..8404)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8878..8894
                     /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 gagcagccaa agctagcctc
     2341 ggccggatct tcccagagtc ctccatcctg ttcctatgtg acatgcagga gaaacttcga
     2401 gatagaatcc tctacttccc acagatcgtg tcaaaggccg ctcggatgct taaggtagcc
     2461 cagatgctag aaatccccgt cttgctgaca gagcattacc cacaaggtct gggccccaca
     2521 gttcccgagc tgggtgctca gggtctgcga accatgagca aaacgtcctt cagcatggtg
     2581 cccccgcttc aacaggagtt ggacaaactg ccccagctgc agtccgtgct actctgtggc
     2641 attgagaccc aaggctgcat cttgcacaca gctctggacc tcctggaccg tgggctacag
     2701 gttcatgtag cagtggatgc ctgttcttct caaagtgaga tgaaccggct cgtggcactg
     2761 gcccgaatgc agcagagtgg cgtctttctg tccacgagcg aggtgctgat tctccagctg
     2821 gtgaaggatg ctgcccaccc acagttcaag gagattcaga agatcctcaa ggagccagtc
     2881 acagacatcg gactgttagg cttcttccaa ggcaagaaaa actccctgtt gcccaactcc
     2941 aggaccggat ccgactacaa agaccatgac ggtgattata aagatcatga catcgactac
     3001 aaggatgacg atgacaagta gcggccgcga aggatctgcg atcgctccgg tgcccgtcag
     3061 tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga
     3121 acgggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc
     3181 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt
     3241 ctttttcgca acgggtttgc cgccagaaca cagctgaagc ttcgaggggc tcgcatctct
     3301 ccttcacgcg cccgccgccc tacctgaggc cgccatccac gccggttgag tcgcgttctg
     3361 ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag tttaaagctc
     3421 aggtcgagac cgggcctttg tccggcgctc ccttggagcc tacctagact cagccggctc
     3481 tccacgcttt gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg
     3541 cgccgttaca gatccaagct gtgaccggcg cctacgctag acgccaccat ggagagcgac
     3601 gagagcggcc tgcccgccat ggagatcgag tgccgcatca ccggcaccct gaacggcgtg
     3661 gagttcgagc tggtgggcgg cggagagggc acccccaagc agggccgcat gaccaacaag
     3721 atgaagagca ccaaaggcgc cctgaccttc agcccctacc tgctgagcca cgtgatgggc
     3781 tacggcttct accacttcgg cacctacccc agcggctacg agaacccctt cctgcacgcc
     3841 atcaacaacg gcggctacac caacacccgc atcgagaagt acgaggacgg cggcgtgctg
     3901 cacgtgagct tcagctaccg ctacgaggcc ggccgcgtga tcggcgactt caaggtggtg
     3961 ggcaccggct tccccgagga cagcgtgatc ttcaccgaca agatcatccg cagcaacgcc
     4021 accgtggagc acctgcaccc catgggcgat aacgtgctgg tgggcagctt cgcccgcacc
     4081 ttcagcctgc gcgacggcgg ctactacagc ttcgtggtgg acagccacat gcacttcaag
     4141 agcgccatcc accccagcat cctgcagaac gggggcccca tgttcgcctt ccgccgcgtg
     4201 gaggagctgc acagcaacac cgagctgggc atcgtggagt accagcacgc cttcaagacc
     4261 cccatcgcct tcgccagatc ccgcgctcag tcgtccaatt ctgccgtgga cggcaccgcc
     4321 ggacccggct ccaccggatc tcgcgagggc agaggaagtc ttctaacatg cggtgacgtg
     4381 gaggagaatc ccggccctat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac
     4441 gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc
     4501 cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc
     4561 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg
     4621 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc
     4681 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg
     4741 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac
     4801 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc
     4861 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc
     4921 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc
     4981 cgcaagcccg gtgcctgaaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     5041 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     5101 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     5161 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     5221 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     5281 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     5341 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg
     5401 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     5461 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     5521 cggcctcttc cgcgtctccg ccttcgccct cagacgagtc ggatctccct ttggccgcct
     5581 ccccgcctgg tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt
     5641 ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaaaata agatctgctt
     5701 tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     5761 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     5821 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     5881 aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag
     5941 aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa
     6001 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     6061 ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc
     6121 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     6181 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     6241 tagacttttg cagagacggc ccaaattcgt aatcatggtc atagctgttt cctgtgtgaa
     6301 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     6361 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     6421 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     6481 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     6541 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     6601 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6661 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6721 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6781 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6841 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6901 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6961 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     7021 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     7081 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     7141 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     7201 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     7261 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     7321 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     7381 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     7441 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     7501 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     7561 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     7621 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7681 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7741 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7801 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7861 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7921 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7981 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     8041 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     8101 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     8161 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     8221 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     8281 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     8341 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     8401 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     8461 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     8521 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     8581 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     8641 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     8701 cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca
     8761 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     8821 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
     8881 aaacgacggc cagtgccaag ctg
//
