LOCUS       .                       9623 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        6..233
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     misc_feature    458..583
                     /label="HIV-1 Ψ"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1076..1309
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             1493..1537
                     /label="gp41 peptide"
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
                     al., 2013)"
     CDS             1686..1727
                     /gene="Protein Tat"
                     /label="Protein Tat"
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
     misc_feature    1804..1921
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2011..2214
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             2303..3460
                     /label="ovalbumin"
                     /note="chicken ovalbumin"
     CDS             3467..3532
                     /label="3xFLAG"
                     /note="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
     promoter        3576..3787
                     /label="EF-1α core promoter"
                     /note="core promoter for human elongation factor EF-1α"
     LTR             3800..4068
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     CDS             4124..4789
                     /label="CopGFP"
                     /note="green fluorescent protein 2 from Pontellina plumata,
                     also known as ppluGFP2 (Shagin et al., 2004)"
     CDS             4859..4912
                     /label="T2A"
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             4919..5710
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     misc_feature    5720..6308
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             6380..6613
                     /label="3' LTR (ΔU3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6685..6819
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      6825..6960
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     primer_bind     complement(6998..7014)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(7022..7038)
                     /label="lac operator"
                     /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)."
     promoter        complement(7046..7076)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7091..7112)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7400..7988)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(8162..9019)
                     /label="AmpR"
                     /note="β-lactamase"
     promoter        complement(9020..9124)
                     /label="AmpR promoter"
     primer_bind     9598..9614
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
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 ccatgggctc catcggtgca gcaagcatgg aattttgttt
     2341 tgatgtattc aaggagctca aagtccacca tgccaatgag aacatcttct actgccccat
     2401 tgccatcatg tcagctctag ccatggtata cctgggtgca aaagacagca ccaggacaca
     2461 aataaataag gttgttcgct ttgataaact tccaggattc ggagacagta ttgaagctca
     2521 gtgtggcaca tctgtaaacg ttcactcttc acttagagac atcctcaacc aaatcaccaa
     2581 accaaatgat gtttattcgt tcagccttgc cagtagactt tatgctgaag agagataccc
     2641 aatcctgcca gaatacttgc agtgtgtgaa ggaactgtat agaggaggct tggaacctat
     2701 caactttcaa acagctgcag atcaagccag agagctcatc aattcctggg tagaaagtca
     2761 gacaaatgga attatcagaa atgtccttca gccaagctcc gtggattctc aaactgcaat
     2821 ggttctggtt aatgccattg tcttcaaagg actgtgggag aaagcattta aggatgaaga
     2881 cacacaagca atgcctttca gagtgactga gcaagaaagc aaacctgtgc agatgatgta
     2941 ccagattggt ttatttagag tggcatcaat ggcttctgag aaaatgaaga tcctggagct
     3001 tccatttgcc agtgggacaa tgagcatgtt ggtgctgttg cctgatgaag tctcaggcct
     3061 tgagcagctt gagagtataa tcaactttga aaaactgact gaatggacca gttctaatgt
     3121 tatggaagag aggaagatca aagtgtactt acctcgcatg aagatggagg aaaaatacaa
     3181 cctcacatct gtcttaatgg ctatgggcat tactgacgtg tttagctctt cagccaatct
     3241 gtctggcatc tcctcagcag agagcctgaa gatatctcaa gctgtccatg cagcacatgc
     3301 agaaatcaat gaagcaggca gagaggtggt agggtcagca gaggctggag tggatgctgc
     3361 aagcgtctct gaagaattta gggctgacca tccattcctc ttctgtatca agcacatcgc
     3421 aaccaacgcc gttctcttct ttggcagatg tgtttcccct ggatccgact acaaagacca
     3481 tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca agtagcggcc
     3541 gcgaaggatc tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt
     3601 ccccgagaag ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg
     3661 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga
     3721 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag
     3781 aacacagctg aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg
     3841 aggccgccat ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg
     3901 aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc
     3961 gctcccttgg agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc
     4021 tcaactctac gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc
     4081 ggcgcctacg ctagacgcca ccatggagag cgacgagagc ggcctgcccg ccatggagat
     4141 cgagtgccgc atcaccggca ccctgaacgg cgtggagttc gagctggtgg gcggcggaga
     4201 gggcaccccc aagcagggcc gcatgaccaa caagatgaag agcaccaaag gcgccctgac
     4261 cttcagcccc tacctgctga gccacgtgat gggctacggc ttctaccact tcggcaccta
     4321 ccccagcggc tacgagaacc ccttcctgca cgccatcaac aacggcggct acaccaacac
     4381 ccgcatcgag aagtacgagg acggcggcgt gctgcacgtg agcttcagct accgctacga
     4441 ggccggccgc gtgatcggcg acttcaaggt ggtgggcacc ggcttccccg aggacagcgt
     4501 gatcttcacc gacaagatca tccgcagcaa cgccaccgtg gagcacctgc accccatggg
     4561 cgataacgtg ctggtgggca gcttcgcccg caccttcagc ctgcgcgacg gcggctacta
     4621 cagcttcgtg gtggacagcc acatgcactt caagagcgcc atccacccca gcatcctgca
     4681 gaacgggggc cccatgttcg ccttccgccg cgtggaggag ctgcacagca acaccgagct
     4741 gggcatcgtg gagtaccagc acgccttcaa gacccccatc gccttcgcca gatcccgcgc
     4801 tcagtcgtcc aattctgccg tggacggcac cgccggaccc ggctccaccg gatctcgcga
     4861 gggcagagga agtcttctaa catgcggtga cgtggaggag aatcccggcc ctgccaccat
     4921 ggctgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     4981 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     5041 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     5101 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     5161 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     5221 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     5281 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     5341 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     5401 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     5461 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     5521 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     5581 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     5641 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     5701 cgagttcttc tgagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     5761 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     5821 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     5881 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     5941 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     6001 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     6061 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     6121 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     6181 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     6241 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     6301 ctccccgcgg tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt
     6361 ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagata agatctgctt
     6421 tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     6481 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     6541 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     6601 aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag
     6661 aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa
     6721 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     6781 ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc
     6841 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     6901 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     6961 tagacttttg cagagacggc ccaaattcgt aatcatggtc atagctgttt cctgtgtgaa
     7021 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     7081 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     7141 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     7201 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     7261 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     7321 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     7381 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     7441 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     7501 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     7561 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     7621 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     7681 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     7741 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     7801 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     7861 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     7921 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     7981 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     8041 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     8101 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     8161 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     8221 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     8281 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     8341 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     8401 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     8461 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     8521 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     8581 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     8641 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     8701 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     8761 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     8821 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     8881 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     8941 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     9001 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     9061 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     9121 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     9181 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     9241 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     9301 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     9361 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     9421 cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca
     9481 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     9541 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
     9601 aaacgacggc cagtgccaag ctg
//
