LOCUS       V036265                 9283 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036265
VERSION     V036265
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..3309
                     /label="Lrrc26(NM_146117)"
                     /note="Lrrc26(NM_146117)"
                     /gene="Lrrc26"
     CDS             3316..3381
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3425..3636
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3649..3917
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3946..3955
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3976..4638
                     /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             4708..4761
                     /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             4762..5361
                     /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    5362..5950
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6024..6257
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6329..6450
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6490..6625
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6658..6674)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6682..6698
                     /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(6706..6736)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6751..6772
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7060..7648)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7819..8679)
                     /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(8680..8784)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9258..9274
                     /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 gccaccatgc ggggttcttt tttctcgcgg
     2341 cttccgccgc aactctctct gctgctgctg ctgtcgttga ggcgagtctg gacccaggag
     2401 gatattggaa ctgccccttc taaatccccg gtggcccccg aatgccccga ggcatgttca
     2461 tgttcactag gcggcaaggc caattgctcc gcactcgcgc tgcctgcggt accagcggac
     2521 ctgagctggc aagtacgctc actgctgctg gatcacaatc gcgtgagcgc gctgcctccg
     2581 ggtgccttcg ccaatgcagg cgcgctgcta tacctagatc tgagggagaa ccggcttcgg
     2641 tcggtgcacg cacgagcttt ctggggtctg ggagtgttgc aatggctgga cctgagctcc
     2701 aaccagctgg aaactctgcc tcctggcacc ttcgcgccgc tgcgcgcgct tagtttcctc
     2761 tccctagcgg gtaaccggct ggcactcctg gagccttcga tcctgggccc gcttccatta
     2821 ctgcgagtgc tcagcctgca ggacaattca ctatcggcaa tcgaggcggg tttgctgaat
     2881 aacttgcctg ccctcgatgt gttgcgcttg catggcaacc cctggacgtg caactgtgcg
     2941 ctgcgtcccc tttgcacttg gctgcgtaag cacccgcgtc cagcctcaga aactgagacc
     3001 ctgctctgcg tgtctccaag actccagacg ctcagcctac tgacagcttt tccggatgcc
     3061 gccttcaaac agtgcactca gtcactagca gcgcgagacc tggcggtggt ctacgctctc
     3121 gggccggtct ctttccttgc tagtttggcc atctgcctgg cattgggctc cgtgctcact
     3181 gcttgtggtg cacggcgccg ccgccgccgc cgcaccacgg tgcgccactt actaaggaga
     3241 cagctagacc ccgagggccc accctccctg gaggatgctg ggagccctgt aacagcagct
     3301 atccaagccg gatccgacta caaagaccat gacggtgatt ataaagatca tgacatcgac
     3361 tacaaggatg acgatgacaa gtagcggccg cgaaggatct gcgatcgctc cggtgcccgt
     3421 cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat
     3481 tgaacgggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg
     3541 ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac
     3601 gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg ggctcgcatc
     3661 tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt
     3721 ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag
     3781 ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg
     3841 ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc gttttctgtt
     3901 ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc tagacgccac catggagagc
     3961 gacgagagcg gcctgcccgc catggagatc gagtgccgca tcaccggcac cctgaacggc
     4021 gtggagttcg agctggtggg cggcggagag ggcaccccca agcagggccg catgaccaac
     4081 aagatgaaga gcaccaaagg cgccctgacc ttcagcccct acctgctgag ccacgtgatg
     4141 ggctacggct tctaccactt cggcacctac cccagcggct acgagaaccc cttcctgcac
     4201 gccatcaaca acggcggcta caccaacacc cgcatcgaga agtacgagga cggcggcgtg
     4261 ctgcacgtga gcttcagcta ccgctacgag gccggccgcg tgatcggcga cttcaaggtg
     4321 gtgggcaccg gcttccccga ggacagcgtg atcttcaccg acaagatcat ccgcagcaac
     4381 gccaccgtgg agcacctgca ccccatgggc gataacgtgc tggtgggcag cttcgcccgc
     4441 accttcagcc tgcgcgacgg cggctactac agcttcgtgg tggacagcca catgcacttc
     4501 aagagcgcca tccaccccag catcctgcag aacgggggcc ccatgttcgc cttccgccgc
     4561 gtggaggagc tgcacagcaa caccgagctg ggcatcgtgg agtaccagca cgccttcaag
     4621 acccccatcg ccttcgccag atcccgcgct cagtcgtcca attctgccgt ggacggcacc
     4681 gccggacccg gctccaccgg atctcgcgag ggcagaggaa gtcttctaac atgcggtgac
     4741 gtggaggaga atcccggccc tatgaccgag tacaagccca cggtgcgcct cgccacccgc
     4801 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     4861 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     4921 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     4981 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     5041 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     5101 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     5161 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     5221 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     5281 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     5341 acccgcaagc ccggtgcctg aaatcaacct ctggattaca aaatttgtga aagattgact
     5401 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     5461 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     5521 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     5581 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     5641 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     5701 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     5761 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     5821 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     5881 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     5941 gcctccccgc ctggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca
     6001 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gataagatct
     6061 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     6121 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     6181 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     6241 gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc
     6301 aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa
     6361 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     6421 tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta
     6481 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     6541 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     6601 tagtgaggag gcttttttgg aggcctagac ttttgcagag accaaattcg taatcatgtc
     6661 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     6721 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     6781 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     6841 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     6901 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     6961 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     7021 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     7081 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     7141 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     7201 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     7261 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     7321 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     7381 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     7441 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     7501 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     7561 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     7621 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     7681 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     7741 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     7801 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7861 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     7921 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     7981 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     8041 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     8101 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     8161 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     8221 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     8281 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     8341 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     8401 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     8461 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     8521 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     8581 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     8641 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     8701 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     8761 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     8821 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     8881 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     8941 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     9001 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     9061 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     9121 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     9181 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     9241 ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctg
//
