LOCUS       .                       9464 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..1920
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2010..2213
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             2302..2952
                     /label="mKO2"
                     /note="monomeric Kusabira-Orange 2 fluorescent protein"
     CDS             3007..3279
                     /label="Cdt1 (30-120)"
                     /note="degron consisting of residues 30-120 of human Cdt1
                     (Zielke and Edgar, 2015)"
     CDS             4054..4383
                     /label="Gem1 (1-110)"
                     /note="degron consisting of residues 1-110 of human Geminin
                     (Zielke and Edgar, 2015)"
     promoter        4434..4645
                     /label="EF-1α core promoter"
                     /note="core promoter for human elongation factor EF-1α"
     LTR             4658..4926
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     CDS             4953..5549
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    5559..6147
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             6221..6454
                     /label="3' LTR (ΔU3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6526..6660
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      6666..6801
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     primer_bind     complement(6839..6855)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(6863..6879)
                     /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(6887..6917)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6932..6953)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7241..7829)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(8003..8860)
                     /label="AmpR"
                     /note="β-lactamase"
     promoter        complement(8861..8965)
                     /label="AmpR promoter"
     primer_bind     9439..9455
                     /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 attacaaaat tcaaaatttt
     1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag
     2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct
     2281 agagctagcg aattcgccac catggtgtcc gtgatcaagc ccgagatgaa gatgcggtac
     2341 tacatggacg gcagcgtgaa cggccacgag ttcaccatcg agggcgaggg caccggcaga
     2401 ccttacgagg gccaccagga aatgaccctg agagtcacca tggccgaggg cggacccatg
     2461 cccttcgcct tcgacctggt gtcccacgtg ttctgctacg gccaccgggt gttcaccaag
     2521 taccccgagg aaatccccga ctacttcaag caggccttcc ccgagggcct gagctgggag
     2581 cggagcctgg aattcgagga tggcggcagc gcctctgtgt ctgcccacat cagcctgcgg
     2641 ggcaacacct tctaccacaa gagcaagttc accggcgtga acttccccgc cgacggcccc
     2701 atcatgcaga accagagcgt ggactgggag cccagcaccg agaagatcac cgccagcgac
     2761 ggcgtgctga agggcgacgt gaccatgtac ctgaagctgg aaggcggcgg aaaccacaag
     2821 tgccagatga aaaccaccta caaggccgcc aaagaaatcc tggaaatgcc cggcgaccac
     2881 tacatcggcc accggctggt gcggaaaacc gagggcaaca tcaccgagca ggtggaggat
     2941 gccgtggccc actacagcat gctgcctagc ctcggcggct accctagcca ttggaggccc
     3001 ctggaaccct ctcctgccag acctgccctg agagcccctg cctccgccac ctccggcagc
     3061 agaaagagag ccagacctcc tgccgcccct ggcagagatc aggccaggcc ccctgcccgc
     3121 agacggctcc ggctgtccgt ggatgaggtg tccagcccta gcacccccga ggcccccgat
     3181 atccctgcct gtcccagccc tggccagaaa atcaagaaga gcacacctgc cgccggacag
     3241 cctcctcacc tgaccagcgc ccaggaccag gacaccatcg tcgacgaggg caggggcagc
     3301 ctgctgacct gcggcgacgt ggaggagaac cccggcccca gatccatggt gtccgtgatc
     3361 aagcccgaga tgaagatcaa gctgtgcatg cggggaaccg tgaacggcca caacttcgtg
     3421 atcgagggcg agggcaaggg aaacccctac gagggcaccc agatcctgga cctgaacgtg
     3481 accgaaggcg cccctctgcc cttcgcctac gacatcctga ccaccgtgtt ccagtacggc
     3541 aaccgggcct tcaccaagta ccccgccgac atccaggact acttcaagca gaccttcccc
     3601 gagggctacc actgggagcg gagcatgacc tacgaggacc agggcatctg caccgccacc
     3661 tccaacatca gcatgagggg cgactgcttt ttctacgaca tcagattcga cggcaccaac
     3721 ttccccccca acggccccgt gatgcagaaa aagaccctga agtgggagcc cagcaccgag
     3781 aagatgtacg tggaggacgg cgtgctgaag ggcgacgtga acatgcggct gctgctggaa
     3841 ggcggcggac actacagatg cgacttcaag accacctaca aggccaagaa agaagtgcgg
     3901 ctgcccgacg cccacaagat cgaccaccgg atcgagatcc tgaagcacga caaggactac
     3961 aacaaagtga agctgtacga gaacgccgtg gccaggtaca gcatgctgcc cagccaggcc
     4021 aagggctacc ccagccactg gcggcctctg gaaatgaacc ccagcatgaa gcagaagcag
     4081 gaagagatca aagagaacat caagaacagc agcgtgccca ggcggaccct gaagatgatc
     4141 cagcccagcg ccagcggatc tctggtgggc cgggagaatg agctgtctgc cggcctgagc
     4201 aagcggaagc accggaacga ccacctgacc agcaccacct ctagcccagg cgtgatcgtg
     4261 cccgagagca gcgagaacaa gaacctgggc ggcgtgaccc aagaaagctt cgacctgatg
     4321 atcaaagaaa accccagcag ccagtactgg aaagaggtgg ccgagaagcg gcggaaggcc
     4381 ctgtagggat ccgcggccgc gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga
     4441 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc
     4501 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt
     4561 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg
     4621 caacgggttt gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg
     4681 cgcccgccgc cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc
     4741 cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag
     4801 accgggcctt tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct
     4861 ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta
     4921 cagatccaag ctgtgaccgg cgcctacgct agatgaccga gtacaagccc acggtgcgcc
     4981 tcgccacccg cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg ttcgccgact
     5041 accccgccac gcgccacacc gtcgacccgg accgccacat cgagcgggtc accgagctgc
     5101 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
     5161 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
     5221 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
     5281 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg
     5341 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
     5401 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
     5461 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
     5521 cctggtgcat gacccgcaag cccggtgcct gagtcgacaa 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 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc
     6001 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     6061 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     6121 ggatctccct ttgggccgcc tccccgcctg gtacctttaa gaccaatgac ttacaaggca
     6181 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc
     6241 caacgaaaat aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga
     6301 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     6361 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     6421 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt
     6481 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca
     6541 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     6601 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc
     6661 tagctatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     6721 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     6781 gaggaggctt ttttggaggc ctagactttt gcagagacgg cccaaattcg taatcatggt
     6841 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     6901 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     6961 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     7021 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     7081 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     7141 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     7201 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     7261 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     7321 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     7381 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     7441 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     7501 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     7561 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     7621 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     7681 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     7741 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     7801 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     7861 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     7921 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     7981 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     8041 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     8101 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     8161 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     8221 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     8281 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     8341 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     8401 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     8461 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     8521 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     8581 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     8641 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     8701 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     8761 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     8821 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     8881 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     8941 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     9001 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     9061 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     9121 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     9181 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     9241 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
     9301 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     9361 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     9421 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctg
//
