LOCUS       V027319                 8773 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027319
VERSION     V027319
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..2799
                     /label="Isg15(NM_015783)"
                     /note="Isg15(NM_015783)"
                     /gene="Isg15"
     CDS             2806..2871
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        2915..3126
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3139..3407
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3436..3445
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3466..4128
                     /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             4198..4251
                     /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             4252..4851
                     /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    4852..5440
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5514..5747
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5819..5940
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5980..6115
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6148..6164)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6172..6188
                     /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(6196..6226)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6241..6262
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6550..7138)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7309..8169)
                     /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(8170..8274)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8748..8764
                     /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 cctgggacct aaaggtgaag
     2341 atgctggggg gtaacgattt cctggtgtcc gtgactaact ccatgacggt gtcagaactg
     2401 aagaagcaga ttgcccagaa gattggtgtg ccggctttcc agcagcgcct ggcccaccaa
     2461 actgcagtgc tccaggacgg tcttaccctt tccagtctgg gtctgggtcc cagcagcaca
     2521 gtgatgctag tggtacagaa ctgcagcgag cctctgagca tcctggtgag gaacgaaagg
     2581 ggccacagca acatctatga ggtctttctg acgcagactg tagacacgct taagaagaag
     2641 gtgtcccagc gggaacaagt ccacgaagac cagttctggc tgagcttcga gggaaggccc
     2701 atggaggaca aggagctgct gggggagtat ggcctaaagc cccagtgcac agtgatcaag
     2761 catttgcgcc tgaggggtgg gggaggggac cagtgtgccg gatccgacta caaagaccat
     2821 gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagcggccg
     2881 cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc
     2941 cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg
     3001 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa
     3061 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga
     3121 acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga
     3181 ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga
     3241 actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg
     3301 ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct
     3361 caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg
     3421 gcgcctacgc tagacgccac catggagagc gacgagagcg gcctgcccgc catggagatc
     3481 gagtgccgca tcaccggcac cctgaacggc gtggagttcg agctggtggg cggcggagag
     3541 ggcaccccca agcagggccg catgaccaac aagatgaaga gcaccaaagg cgccctgacc
     3601 ttcagcccct acctgctgag ccacgtgatg ggctacggct tctaccactt cggcacctac
     3661 cccagcggct acgagaaccc cttcctgcac gccatcaaca acggcggcta caccaacacc
     3721 cgcatcgaga agtacgagga cggcggcgtg ctgcacgtga gcttcagcta ccgctacgag
     3781 gccggccgcg tgatcggcga cttcaaggtg gtgggcaccg gcttccccga ggacagcgtg
     3841 atcttcaccg acaagatcat ccgcagcaac gccaccgtgg agcacctgca ccccatgggc
     3901 gataacgtgc tggtgggcag cttcgcccgc accttcagcc tgcgcgacgg cggctactac
     3961 agcttcgtgg tggacagcca catgcacttc aagagcgcca tccaccccag catcctgcag
     4021 aacgggggcc ccatgttcgc cttccgccgc gtggaggagc tgcacagcaa caccgagctg
     4081 ggcatcgtgg agtaccagca cgccttcaag acccccatcg ccttcgccag atcccgcgct
     4141 cagtcgtcca attctgccgt ggacggcacc gccggacccg gctccaccgg atctcgcgag
     4201 ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc tatgaccgag
     4261 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc
     4321 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc
     4381 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     4441 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     4501 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     4561 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     4621 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     4681 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc
     4741 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     4801 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg aaatcaacct
     4861 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     4921 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     4981 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     5041 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     5101 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     5161 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     5221 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
     5281 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     5341 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     5401 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt taagaccaat
     5461 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
     5521 gctaattcac tcccaacgaa gataagatct gctttttgct tgtactgggt ctctctggtt
     5581 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     5641 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     5701 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg
     5761 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa
     5821 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     5881 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5941 tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc
     6001 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     6061 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac
     6121 ttttgcagag accaaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc
     6181 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     6241 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     6301 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6361 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6421 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6481 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6541 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6601 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6661 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6721 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6781 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6841 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6901 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6961 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     7021 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     7081 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     7141 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     7201 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     7261 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     7321 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7381 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7441 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7501 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7561 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7621 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7681 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7741 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7801 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7861 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7921 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7981 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     8041 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     8101 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     8161 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     8221 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     8281 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     8341 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     8401 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     8461 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     8521 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     8581 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
     8641 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     8701 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     8761 cagtgccaag ctg
//
