LOCUS       V035880                 9463 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035880
VERSION     V035880
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..3577
                     /label="UBR7(NM_175748)"
                     /note="UBR7(NM_175748)"
                     /gene="UBR7"
     promoter        3622..3833
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3846..4114
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4143..4152
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4173..4835
                     /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             4905..4958
                     /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             4959..5558
                     /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    5559..6146
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6220..6453
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6525..6646
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6665..6800
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6838..6854)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6862..6878
                     /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(6886..6916)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6931..6952
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7240..7828)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7999..8859)
                     /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(8860..8964)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9438..9454
                     /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 ccatggccgg agccgagggc gccgctgggc ggcagtcgga
     2341 gctggagccc gtggtatcgt tggtcgacgt ccttgaggag gacgaggagc tggagaatga
     2401 ggcgtgcgct gtcctgggcg gcagcgactc cgagaagtgc tcctactctc agggctcagt
     2461 aaagagacaa gcactatatg cctgtagtac ctgcacccca gagggagaag aaccagcagg
     2521 aatttgttta gcttgcagtt atgaatgtca tggaagtcac aaactatttg agctatacac
     2581 aaaaagaaat tttcgttgtg attgtggaaa cagcaagttt aaaaatttgg aatgcaaatt
     2641 acttcctgac aaagcaaagg taaattctgg caataagtac aatgacaact tttttggatt
     2701 gtactgcatt tgcaagagac cttatcctga tcctgaagac gagattccag atgagatgat
     2761 ccagtgcgta gtctgtgaag actggttcca tggaaggcat cttggtgcca ttccccctga
     2821 gagtggggat tttcaggaga tggtatgcca ggcctgcatg aaacgttgtt cttttttgtg
     2881 ggcttatgct gcacaattgg cagtaaccaa aatatccact gaggatgatg gattggtgcg
     2941 gaacattgat ggaataggtg atcaggaagt tatcaaacct gaaaatggag agcatcaaga
     3001 tagtaccctc aaagaggatg ttccagaaca gggaaaggat gatgtccggg aggttaaagt
     3061 agagcagaac agtgaaccat gtgccggctc tagttctgaa tctgatctcc agacagtgtt
     3121 taagaatgaa agcctcaacg cagaatcaaa atctggctgc aaacttcagg agcttaaagc
     3181 taagcagctt ataaagaaag acactgccac ctattggccc ctgaactggc gtagcaagtt
     3241 gtgtacctgc caagactgta tgaaaatgta tggagatcta gatgtcttat tcctgacaga
     3301 tgaatacgac acagttctgg cttatgaaaa caaagggaag attgcccagg ccactgacag
     3361 gagcgatccc ctaatggata cccttagcag catgaataga gtccagcaag tggaactcat
     3421 ttgtgaatac aatgatttga agactgaact taaagactat ctcaagagat ttgctgatga
     3481 aggcacggtt gttaagagag aggacattca gcagttcttt gaagagtttc agtcaaaaaa
     3541 gagaagaaga gtggatggga tgcagtatta ctgcagctag gcggccgcga aggatctgcg
     3601 atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
     3661 ggggaggggt cggcaattga acgggtgcct agagaaggtg gcgcggggta aactgggaaa
     3721 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
     3781 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca cagctgaagc
     3841 ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc cgccatccac
     3901 gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg
     3961 tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc ccttggagcc
     4021 tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa ctctacgtct
     4081 ttgtttcgtt ttctgttctg cgccgttaca gatccaagct gtgaccggcg cctacgctag
     4141 acgccaccat ggagagcgac gagagcggcc tgcccgccat ggagatcgag tgccgcatca
     4201 ccggcaccct gaacggcgtg gagttcgagc tggtgggcgg cggagagggc acccccaagc
     4261 agggccgcat gaccaacaag atgaagagca ccaaaggcgc cctgaccttc agcccctacc
     4321 tgctgagcca cgtgatgggc tacggcttct accacttcgg cacctacccc agcggctacg
     4381 agaacccctt cctgcacgcc atcaacaacg gcggctacac caacacccgc atcgagaagt
     4441 acgaggacgg cggcgtgctg cacgtgagct tcagctaccg ctacgaggcc ggccgcgtga
     4501 tcggcgactt caaggtggtg ggcaccggct tccccgagga cagcgtgatc ttcaccgaca
     4561 agatcatccg cagcaacgcc accgtggagc acctgcaccc catgggcgat aacgtgctgg
     4621 tgggcagctt cgcccgcacc ttcagcctgc gcgacggcgg ctactacagc ttcgtggtgg
     4681 acagccacat gcacttcaag agcgccatcc accccagcat cctgcagaac gggggcccca
     4741 tgttcgcctt ccgccgcgtg gaggagctgc acagcaacac cgagctgggc atcgtggagt
     4801 accagcacgc cttcaagacc cccatcgcct tcgccagatc ccgcgctcag tcgtccaatt
     4861 ctgccgtgga cggcaccgcc ggacccggct ccaccggatc tcgcgagggc agaggaagtc
     4921 ttctaacatg cggtgacgtg gaggagaatc ccggccctat gaccgagtac aagcccacgg
     4981 tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg
     5041 ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg
     5101 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg
     5161 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt
     5221 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac
     5281 agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg
     5341 tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag
     5401 tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc
     5461 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac
     5521 cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgaaa tcaacctctg gattacaaaa
     5581 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     5641 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     5701 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     5761 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     5821 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     5881 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     5941 tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc
     6001 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     6061 gcggcctgct gccggctctg cggcctcttc cgcgtctccg ccttcgccct cagacgagtc
     6121 ggatctccct ttggccgcct ccccgcctgg tacctttaag accaatgact tacaaggcag
     6181 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc
     6241 aacgaaaata agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag
     6301 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     6361 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     6421 gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc
     6481 agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag
     6541 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     6601 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct
     6661 agctatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     6721 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     6781 aggaggcttt tttggaggcc tagacttttg cagagacggc ccaaattcgt aatcatggtc
     6841 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     6901 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     6961 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     7021 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     7081 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     7141 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     7201 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     7261 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     7321 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     7381 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     7441 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     7501 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     7561 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     7621 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     7681 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     7741 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     7801 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     7861 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     7921 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     7981 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     8041 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     8101 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     8161 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     8221 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     8281 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     8341 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     8401 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     8461 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     8521 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     8581 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     8641 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     8701 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     8761 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     8821 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     8881 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     8941 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     9001 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     9061 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     9121 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     9181 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     9241 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     9301 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     9361 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     9421 ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctg
//
