LOCUS       V024625                 9383 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024625
VERSION     V024625
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..3426
                     /label="Usp12(NM_011669)"
                     /note="Usp12(NM_011669)"
                     /gene="Usp12"
     CDS             3433..3498
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3542..3753
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3766..4034
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4063..4072
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4093..4755
                     /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             4825..4878
                     /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             4879..5478
                     /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    5479..6066
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6140..6373
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6445..6566
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6585..6720
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6758..6774)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6782..6798
                     /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(6806..6836)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6851..6872
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7160..7748)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7919..8779)
                     /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(8780..8884)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9358..9374
                     /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 aaatcctaat gacagtctcc
     2341 aaattcgcct ccatctgtac catgggcgcc aatgcctcgg cattagagaa agagattggt
     2401 ccagaacagt ttccggtcaa cgagcactat tttggattag tcaattttgg gaatacctgc
     2461 tactgcaact cagttcttca agcactttat ttttgtcgtc catttcggga aaaagttcta
     2521 gcatacaaaa gtcagcccag aaaaaaggag aaccttctca cctgcctggc agacctcttc
     2581 cacagcatag ccacccagaa gaagaaggtc ggcgtgatcc cgcccaagaa gttcatcaca
     2641 aggttgcgga aagaaaacga gctgtttgac aactacatgc aacaagatgc ccatgaattc
     2701 ttaaattacc tactcaatac aattgctgat attctacaag aagaaagaaa acaggagaag
     2761 caaaatggcc gattacggaa cggagacgtg gacaacgagg ataacaacag cacaccagac
     2821 ccaacctggg tccacgagat ctttcaggga acattaacta atgaaaccag gtgtcttact
     2881 tgtgaaacta taagcagcaa agatgaagat tttttagacc tttctgttga cgtggaacaa
     2941 aacacatcaa ttactcactg cttaagaggg ttcagtaaca cagaaactct gtgcagtgaa
     3001 tataagtact actgtgaaga gtgccggagc aagcaggaag cacacaagcg gatgaaggtt
     3061 aagaaactgc ccctgatcct ggctctgcac ctgaagaggt ttaagtatat ggaccagctt
     3121 caccggtaca cgaagctttc ttatcgcgtg gtctttcctt tagagctccg tctgttcaac
     3181 acgtcaggag atgcgaccaa ccctgacaga atgtatgacc tggtcgccgt tgtggttcac
     3241 tgtggaagtg gtcccaaccg aggccattat attgcaatag ttaagagtca tgatttctgg
     3301 ctgttgtttg atgatgacat tgtagaaaaa atcgatgccc aagctattga agaattctac
     3361 ggattgacat ctgacatctc caagaactcg gagtctggct acatcctttt ctatcagtct
     3421 cgggacggat ccgactacaa agaccatgac ggtgattata aagatcatga catcgactac
     3481 aaggatgacg atgacaagta gcggccgcga aggatctgcg atcgctccgg tgcccgtcag
     3541 tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga
     3601 acgggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc
     3661 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt
     3721 ctttttcgca acgggtttgc cgccagaaca cagctgaagc ttcgaggggc tcgcatctct
     3781 ccttcacgcg cccgccgccc tacctgaggc cgccatccac gccggttgag tcgcgttctg
     3841 ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag tttaaagctc
     3901 aggtcgagac cgggcctttg tccggcgctc ccttggagcc tacctagact cagccggctc
     3961 tccacgcttt gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg
     4021 cgccgttaca gatccaagct gtgaccggcg cctacgctag acgccaccat ggagagcgac
     4081 gagagcggcc tgcccgccat ggagatcgag tgccgcatca ccggcaccct gaacggcgtg
     4141 gagttcgagc tggtgggcgg cggagagggc acccccaagc agggccgcat gaccaacaag
     4201 atgaagagca ccaaaggcgc cctgaccttc agcccctacc tgctgagcca cgtgatgggc
     4261 tacggcttct accacttcgg cacctacccc agcggctacg agaacccctt cctgcacgcc
     4321 atcaacaacg gcggctacac caacacccgc atcgagaagt acgaggacgg cggcgtgctg
     4381 cacgtgagct tcagctaccg ctacgaggcc ggccgcgtga tcggcgactt caaggtggtg
     4441 ggcaccggct tccccgagga cagcgtgatc ttcaccgaca agatcatccg cagcaacgcc
     4501 accgtggagc acctgcaccc catgggcgat aacgtgctgg tgggcagctt cgcccgcacc
     4561 ttcagcctgc gcgacggcgg ctactacagc ttcgtggtgg acagccacat gcacttcaag
     4621 agcgccatcc accccagcat cctgcagaac gggggcccca tgttcgcctt ccgccgcgtg
     4681 gaggagctgc acagcaacac cgagctgggc atcgtggagt accagcacgc cttcaagacc
     4741 cccatcgcct tcgccagatc ccgcgctcag tcgtccaatt ctgccgtgga cggcaccgcc
     4801 ggacccggct ccaccggatc tcgcgagggc agaggaagtc ttctaacatg cggtgacgtg
     4861 gaggagaatc ccggccctat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac
     4921 gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc
     4981 cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc
     5041 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg
     5101 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc
     5161 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg
     5221 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac
     5281 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc
     5341 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc
     5401 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc
     5461 cgcaagcccg gtgcctgaaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     5521 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     5581 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     5641 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     5701 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     5761 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     5821 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg
     5881 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     5941 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     6001 cggcctcttc cgcgtctccg ccttcgccct cagacgagtc ggatctccct ttggccgcct
     6061 ccccgcctgg tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt
     6121 ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaaaata agatctgctt
     6181 tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     6241 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     6301 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     6361 aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag
     6421 aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa
     6481 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     6541 ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc
     6601 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     6661 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     6721 tagacttttg cagagacggc ccaaattcgt aatcatggtc atagctgttt cctgtgtgaa
     6781 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     6841 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     6901 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     6961 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     7021 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     7081 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     7141 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     7201 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     7261 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     7321 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     7381 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     7441 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     7501 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     7561 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     7621 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     7681 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     7741 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     7801 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     7861 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     7921 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     7981 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     8041 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     8101 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     8161 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     8221 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     8281 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     8341 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     8401 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     8461 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     8521 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     8581 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     8641 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     8701 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     8761 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     8821 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     8881 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     8941 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     9001 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     9061 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     9121 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     9181 cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca
     9241 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     9301 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
     9361 aaacgacggc cagtgccaag ctg
//
