LOCUS       V025735                 9103 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025735
VERSION     V025735
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..3217
                     /label="CA5A(NM_001739)"
                     /note="CA5A(NM_001739)"
                     /gene="CA5A"
     promoter        3262..3473
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3486..3754
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3783..3792
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3813..4475
                     /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             4545..4598
                     /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             4599..5198
                     /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    5199..5786
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5860..6093
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6165..6286
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6305..6440
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6478..6494)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6502..6518
                     /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(6526..6556)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6571..6592
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6880..7468)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7639..8499)
                     /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(8500..8604)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9078..9094
                     /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 ccatgttggg gaggaacact tggaagacct cagctttctc
     2341 cttcttggtt gagcagatgt gggcccctct ctggagtcgt tcgatgaggc cagggcgatg
     2401 gtgttctcag cgttcctgtg catggcaaac cagcaataac actttgcacc cactctggac
     2461 ggtcccggtc tccgtgccag ggggcacccg gcagtctcct attaacatcc agtggaggga
     2521 cagcgtctat gacccccagc tgaagccact cagggtctcc tatgaagcgg catcctgcct
     2581 gtacatctgg aacactggct acctcttcca ggtggaattt gacgatgcca ccgaggcatc
     2641 aggaattagt ggtgggccct tggaaaacca ctacagactg aagcaatttc acttccactg
     2701 gggagcagtg aacgaggggg gctcagagca cacagtggac ggccacgcgt accccgcaga
     2761 gctgcattta gttcactgga attctgtgaa ataccaaaat tacaaggaag ctgtcgtggg
     2821 agagaatggt ttggctgtga taggcgtgtt tttaaagctc ggggcccatc atcagacgct
     2881 gcagaggctg gtggacatct tgccggaaat aaaacataag gacgcgcggg cggccatgcg
     2941 ccccttcgac ccctccactc tgctgcccac ctgctgggat tactggacct acgcgggctc
     3001 gctcaccacc ccgccgctga ccgagtcggt cacctggatc atccagaagg agcccgttga
     3061 agtggcccca agccagctct ctgcatttcg tactctcctg ttttctgcac ttggtgaaga
     3121 ggagaagatg atggtgaaca actatcgccc acttcaaccc ttgatgaacc ggaaggtctg
     3181 ggcgtccttc caggccacta atgagggcac aaggtcctag gcggccgcga aggatctgcg
     3241 atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
     3301 ggggaggggt cggcaattga acgggtgcct agagaaggtg gcgcggggta aactgggaaa
     3361 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
     3421 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca cagctgaagc
     3481 ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc cgccatccac
     3541 gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg
     3601 tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc ccttggagcc
     3661 tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa ctctacgtct
     3721 ttgtttcgtt ttctgttctg cgccgttaca gatccaagct gtgaccggcg cctacgctag
     3781 acgccaccat ggagagcgac gagagcggcc tgcccgccat ggagatcgag tgccgcatca
     3841 ccggcaccct gaacggcgtg gagttcgagc tggtgggcgg cggagagggc acccccaagc
     3901 agggccgcat gaccaacaag atgaagagca ccaaaggcgc cctgaccttc agcccctacc
     3961 tgctgagcca cgtgatgggc tacggcttct accacttcgg cacctacccc agcggctacg
     4021 agaacccctt cctgcacgcc atcaacaacg gcggctacac caacacccgc atcgagaagt
     4081 acgaggacgg cggcgtgctg cacgtgagct tcagctaccg ctacgaggcc ggccgcgtga
     4141 tcggcgactt caaggtggtg ggcaccggct tccccgagga cagcgtgatc ttcaccgaca
     4201 agatcatccg cagcaacgcc accgtggagc acctgcaccc catgggcgat aacgtgctgg
     4261 tgggcagctt cgcccgcacc ttcagcctgc gcgacggcgg ctactacagc ttcgtggtgg
     4321 acagccacat gcacttcaag agcgccatcc accccagcat cctgcagaac gggggcccca
     4381 tgttcgcctt ccgccgcgtg gaggagctgc acagcaacac cgagctgggc atcgtggagt
     4441 accagcacgc cttcaagacc cccatcgcct tcgccagatc ccgcgctcag tcgtccaatt
     4501 ctgccgtgga cggcaccgcc ggacccggct ccaccggatc tcgcgagggc agaggaagtc
     4561 ttctaacatg cggtgacgtg gaggagaatc ccggccctat gaccgagtac aagcccacgg
     4621 tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg
     4681 ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg
     4741 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg
     4801 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt
     4861 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac
     4921 agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg
     4981 tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag
     5041 tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc
     5101 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac
     5161 cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgaaa tcaacctctg gattacaaaa
     5221 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     5281 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     5341 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     5401 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     5461 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     5521 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     5581 tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc
     5641 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     5701 gcggcctgct gccggctctg cggcctcttc cgcgtctccg ccttcgccct cagacgagtc
     5761 ggatctccct ttggccgcct ccccgcctgg tacctttaag accaatgact tacaaggcag
     5821 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc
     5881 aacgaaaata agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag
     5941 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     6001 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     6061 gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc
     6121 agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag
     6181 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     6241 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct
     6301 agctatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     6361 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     6421 aggaggcttt tttggaggcc tagacttttg cagagacggc ccaaattcgt aatcatggtc
     6481 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     6541 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     6601 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     6661 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     6721 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     6781 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     6841 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     6901 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6961 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     7021 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     7081 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     7141 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     7201 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     7261 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     7321 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     7381 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     7441 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     7501 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     7561 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     7621 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7681 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     7741 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     7801 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     7861 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     7921 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     7981 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     8041 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     8101 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     8161 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     8221 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     8281 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     8341 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     8401 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     8461 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     8521 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     8581 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     8641 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     8701 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     8761 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     8821 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     8881 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     8941 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     9001 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     9061 ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctg
//
