LOCUS       V022179                 9513 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022179
VERSION     V022179
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             2303..3628
                     /label="TGFB2(NM_001135599)"
                     /note="TGFB2(NM_001135599)"
                     /gene="TGFB2"
     promoter        3672..3883
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3896..4164
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4193..4202
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4223..4885
                     /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             4955..5008
                     /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             5009..5608
                     /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    5609..6196
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6270..6503
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6575..6696
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6715..6850
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6888..6904)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6912..6928
                     /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(6936..6966)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6981..7002
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7290..7878)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8049..8909)
                     /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(8910..9014)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9488..9504
                     /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 gaattcgcca ccatgcacta ctgtgtgctg agcgcttttc tgatcctgca
     2341 tctggtcacg gtcgcgctca gcctgtctac ctgcagcaca ctcgatatgg accagttcat
     2401 gcgcaagagg atcgaggcga tccgcgggca gatcctgagc aagctgaagc tcaccagtcc
     2461 cccagaagac tatcctgagc ccgaggaagt ccccccggag gtgatttcca tctacaacag
     2521 caccagggac ttgctccagg agaaggcgag ccggagggcg gccgcctgcg agcgcgagag
     2581 gagcgacgaa gagtactacg ccaaggaggt ttacaaaata gacatgccgc ccttcttccc
     2641 ctccgaaact gtctgcccag ttgttacaac accctctggc tcagtgggca gcttgtgctc
     2701 cagacagtcc caggtgctct gtgggtacct tgatgccatc ccgcccactt tctacagacc
     2761 ctacttcaga attgttcgat ttgacgtctc agcaatggag aagaatgctt ccaatttggt
     2821 gaaagcagag ttcagagtct ttcgtttgca gaacccaaaa gccagagtgc ctgaacaacg
     2881 gattgagcta tatcagattc tcaagtccaa agatttaaca tctccaaccc agcgctacat
     2941 cgacagcaaa gttgtgaaaa caagagcaga aggcgaatgg ctctccttcg atgtaactga
     3001 tgctgttcat gaatggcttc accataaaga caggaacctg ggatttaaaa taagcttaca
     3061 ctgtccctgc tgcacttttg taccatctaa taattacatc atcccaaata aaagtgaaga
     3121 actagaagca agatttgcag gtattgatgg cacctccaca tataccagtg gtgatcagaa
     3181 aactataaag tccactagga aaaaaaacag tgggaagacc ccacatctcc tgctaatgtt
     3241 attgccctcc tacagacttg agtcacaaca gaccaaccgg cggaagaagc gtgctttgga
     3301 tgcggcctat tgctttagaa atgtgcagga taattgctgc ctacgtccac tttacattga
     3361 tttcaagagg gatctagggt ggaaatggat acacgaaccc aaagggtaca atgccaactt
     3421 ctgtgctgga gcatgcccgt atttatggag ttcagacact cagcacagca gggtcctgag
     3481 cttatataat accataaatc cagaagcatc tgcttctcct tgctgcgtgt cccaagattt
     3541 agaacctcta accattctct actacattgg caaaacaccc aagattgaac agctttctaa
     3601 tatgattgta aagtcttgca aatgcagcta agcggccgca aggatctgcg atcgctccgg
     3661 tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt
     3721 cggcaattga acgggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt
     3781 gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc
     3841 cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca cagctgaagc ttcgaggggc
     3901 tcgcatctct ccttcacgcg cccgccgccc tacctgaggc cgccatccac gccggttgag
     3961 tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag
     4021 tttaaagctc aggtcgagac cgggcctttg tccggcgctc ccttggagcc tacctagact
     4081 cagccggctc tccacgcttt gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt
     4141 ttctgttctg cgccgttaca gatccaagct gtgaccggcg cctacgctag acgccaccat
     4201 ggagagcgac gagagcggcc tgcccgccat ggagatcgag tgccgcatca ccggcaccct
     4261 gaacggcgtg gagttcgagc tggtgggcgg cggagagggc acccccaagc agggccgcat
     4321 gaccaacaag atgaagagca ccaaaggcgc cctgaccttc agcccctacc tgctgagcca
     4381 cgtgatgggc tacggcttct accacttcgg cacctacccc agcggctacg agaacccctt
     4441 cctgcacgcc atcaacaacg gcggctacac caacacccgc atcgagaagt acgaggacgg
     4501 cggcgtgctg cacgtgagct tcagctaccg ctacgaggcc ggccgcgtga tcggcgactt
     4561 caaggtggtg ggcaccggct tccccgagga cagcgtgatc ttcaccgaca agatcatccg
     4621 cagcaacgcc accgtggagc acctgcaccc catgggcgat aacgtgctgg tgggcagctt
     4681 cgcccgcacc ttcagcctgc gcgacggcgg ctactacagc ttcgtggtgg acagccacat
     4741 gcacttcaag agcgccatcc accccagcat cctgcagaac gggggcccca tgttcgcctt
     4801 ccgccgcgtg gaggagctgc acagcaacac cgagctgggc atcgtggagt accagcacgc
     4861 cttcaagacc cccatcgcct tcgccagatc ccgcgctcag tcgtccaatt ctgccgtgga
     4921 cggcaccgcc ggacccggct ccaccggatc tcgcgagggc agaggaagtc ttctaacatg
     4981 cggtgacgtg gaggagaatc ccggccctat gaccgagtac aagcccacgg tgcgcctcgc
     5041 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     5101 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga
     5161 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     5221 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     5281 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     5341 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     5401 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     5461 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     5521 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     5581 gtgcatgacc cgcaagcccg gtgcctgaaa tcaacctctg gattacaaaa tttgtgaaag
     5641 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     5701 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     5761 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     5821 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     5881 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     5941 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     6001 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     6061 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     6121 gccggctctg cggcctcttc cgcgtctccg ccttcgccct cagacgagtc ggatctccct
     6181 ttggccgcct ccccgcctgg tacctttaag accaatgact tacaaggcag ctgtagatct
     6241 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaaaata
     6301 agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct
     6361 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     6421 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     6481 gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata
     6541 acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg
     6601 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     6661 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg
     6721 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     6781 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     6841 tttggaggcc tagacttttg cagagacggc ccaaattcgt aatcatggtc atagctgttt
     6901 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     6961 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     7021 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     7081 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     7141 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     7201 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     7261 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     7321 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     7381 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     7441 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     7501 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     7561 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     7621 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     7681 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     7741 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     7801 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     7861 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     7921 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     7981 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     8041 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     8101 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     8161 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     8221 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     8281 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     8341 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     8401 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     8461 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     8521 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     8581 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     8641 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     8701 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     8761 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     8821 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     8881 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     8941 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     9001 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     9061 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     9121 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     9181 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     9241 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
     9301 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc
     9361 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     9421 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     9481 cgacgttgta aaacgacggc cagtgccaag ctg
//
