LOCUS       V022779                 9142 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022779
VERSION     V022779
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    1798..1915
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2005..2208
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2297..3184
                     /label="GLYAT(NM_201648)"
                     /note="GLYAT(NM_201648)"
                     /gene="GLYAT"
     CDS             3191..3256
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3301..3512
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3525..3793
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3822..3831
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3852..4514
                     /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             4584..4637
                     /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             4638..5237
                     /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    5238..5825
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5899..6132
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6204..6325
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6344..6479
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6517..6533)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6541..6557
                     /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(6565..6595)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6610..6631
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6919..7507)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7678..8538)
                     /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(8539..8643)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9117..9133
                     /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 ggttaacttt
     1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga
     1921 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
     2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc
     2281 tagcgaattc gccaccatga tgttaccatt gcaaggtgcc cagatgctgc agatgctgga
     2341 gaaatccttg aggaagagcc tcccagcatc cttaaaggtt tatggaactg tctttcacat
     2401 aaaccatgga aatccattca atctgaaggc tgtggtggac aagtggcctg attttaatac
     2461 agtggttgtc tgccctcagg agcaggatat gacagatgac cttgatcact ataccaatac
     2521 ttaccaaatc tactccaaag atccccaaaa ctgtcaggaa ttccttggat caccagaact
     2581 catcaactgg aaacagcatt tacagattca aagttcacag cctagcctga atgaggctat
     2641 acaaaatctt gcagccatta agtccttcaa agtcaaacaa acacaacgca ttctctatat
     2701 ggcagctgaa acagccaagg aactgactcc tttcctgctg aaatcaaaga ttttatctcc
     2761 caatggtggc aaacccaagg ccatcaacca agagatgttt aaactctcat ccatggatgt
     2821 tacccatgct cacttggtga ataaattctg gcattttggt ggtaatgaga ggagccagag
     2881 attcattgag cgctgcattc agacctttcc cacctgctgt ctcctggggc ctgaggggac
     2941 ccctgtgtgc tgggatctaa tggaccagac tggagagatg agaatggcag gcaccttgcc
     3001 ggaataccgg ctccatggcc ttgtgacgta tgtcatctat tcccacgccc agaaattggg
     3061 caaacttggg tttcctgtct attctcatgt agactacagc aatgaagcta tgcaaaaaat
     3121 gagttacaca ctgcaacatg ttcccattcc cagaagctgg aaccagtgga actgtgtacc
     3181 tctgggatcc gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa
     3241 ggatgacgat gacaagtgag cggccgcgaa ggatctgcga tcgctccggt gcccgtcagt
     3301 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa
     3361 cgggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
     3421 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc
     3481 tttttcgcaa cgggtttgcc gccagaacac agctgaagct tcgaggggct cgcatctctc
     3541 cttcacgcgc ccgccgccct acctgaggcc gccatccacg ccggttgagt cgcgttctgc
     3601 cgcctcccgc ctgtggtgcc tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca
     3661 ggtcgagacc gggcctttgt ccggcgctcc cttggagcct acctagactc agccggctct
     3721 ccacgctttg cctgaccctg cttgctcaac tctacgtctt tgtttcgttt tctgttctgc
     3781 gccgttacag atccaagctg tgaccggcgc ctacgctaga cgccaccatg gagagcgacg
     3841 agagcggcct gcccgccatg gagatcgagt gccgcatcac cggcaccctg aacggcgtgg
     3901 agttcgagct ggtgggcggc ggagagggca cccccaagca gggccgcatg accaacaaga
     3961 tgaagagcac caaaggcgcc ctgaccttca gcccctacct gctgagccac gtgatgggct
     4021 acggcttcta ccacttcggc acctacccca gcggctacga gaaccccttc ctgcacgcca
     4081 tcaacaacgg cggctacacc aacacccgca tcgagaagta cgaggacggc ggcgtgctgc
     4141 acgtgagctt cagctaccgc tacgaggccg gccgcgtgat cggcgacttc aaggtggtgg
     4201 gcaccggctt ccccgaggac agcgtgatct tcaccgacaa gatcatccgc agcaacgcca
     4261 ccgtggagca cctgcacccc atgggcgata acgtgctggt gggcagcttc gcccgcacct
     4321 tcagcctgcg cgacggcggc tactacagct tcgtggtgga cagccacatg cacttcaaga
     4381 gcgccatcca ccccagcatc ctgcagaacg ggggccccat gttcgccttc cgccgcgtgg
     4441 aggagctgca cagcaacacc gagctgggca tcgtggagta ccagcacgcc ttcaagaccc
     4501 ccatcgcctt cgccagatcc cgcgctcagt cgtccaattc tgccgtggac ggcaccgccg
     4561 gacccggctc caccggatct cgcgagggca gaggaagtct tctaacatgc ggtgacgtgg
     4621 aggagaatcc cggccctatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg
     4681 acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc
     4741 acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca
     4801 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg
     4861 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca
     4921 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc
     4981 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc
     5041 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg
     5101 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg
     5161 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc
     5221 gcaagcccgg tgcctgaaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     5281 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     5341 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     5401 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     5461 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     5521 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     5581 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt
     5641 cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     5701 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     5761 ggcctcttcc gcgtctccgc cttcgccctc agacgagtcg gatctccctt tggccgcctc
     5821 cccgcctggt acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
     5881 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaaaataa gatctgcttt
     5941 ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     6001 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     6061 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     6121 aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga
     6181 aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt tacaaataaa
     6241 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     6301 tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc ccctaactcc
     6361 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     6421 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     6481 agacttttgc agagacggcc caaattcgta atcatggtca tagctgtttc ctgtgtgaaa
     6541 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     6601 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     6661 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     6721 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6781 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6841 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6901 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6961 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     7021 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     7081 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     7141 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     7201 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     7261 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     7321 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     7381 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     7441 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     7501 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     7561 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     7621 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     7681 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7741 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7801 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7861 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7921 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7981 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     8041 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     8101 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     8161 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     8221 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     8281 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     8341 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     8401 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     8461 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     8521 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     8581 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     8641 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     8701 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     8761 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     8821 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     8881 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     8941 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     9001 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     9061 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     9121 aacgacggcc agtgccaagc tg
//
