LOCUS       .                       9337 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     polyA_signal    complement(216..350)
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     promoter        398..502
                     /label="AmpR promoter"
     CDS             503..1360
                     /label="AmpR"
                     /note="β-lactamase"
     rep_origin      1534..2119
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    2407..2428
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2443..2473
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    2481..2497
                     /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)."
     primer_bind     2505..2521
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     LTR             complement(2650..3283)
                     /label="5' LTR"
                     /note="5' long terminal repeat (LTR) from HIV-1"
     misc_feature    complement(3490..4078)
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     CDS             complement(4095..4691)
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     promoter        complement(4700..5029)
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             complement(5043..5786)
                     /label="Tet-On® 3G"
                     /note="modified rtTA protein that binds tightly to
                     promoters containing the tet operator in the presence of
                     doxycycline"
     promoter        complement(5805..6315)
                     /label="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
     promoter        6332..6699
                     /label="TRE3GS promoter"
                     /note="3rd-generation Tet-responsive promoter that can be
                     activated by binding of Tet-On® 3G, modified to eliminate
                     binding sites for endogenous mammalian transcription
                     factors"
     CDS             6732..6797
                     /label="3xFLAG"
                     /note="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
     CDS             6804..6830
                     /label="9xHis"
                     /note="9xHis affinity tag"
     misc_feature    complement(7194..7311)
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     CDS             complement(7383..7424)
                     /gene="Protein Tat"
                     /label="Protein Tat"
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
     CDS             complement(7573..7617)
                     /label="gp41 peptide"
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
                     al., 2013)"
     misc_feature    complement(7802..8035)
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     misc_feature    complement(8532..8657)
                     /label="HIV-1 Ψ"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     LTR             complement(8704..9337)
                     /label="3' LTR"
                     /note="3' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 actactaagt ttgtagtaca tatttaacaa atacaatttc tttaaaaact actaagtttg
       61 tagtacatat ttaacaaata caatttcttt aaaatgaaaa taattcagag gaatcacagg
      121 tttagagtaa atgaaaccac taggtaattg gcagtggtaa tagggtatgg ggtgggaagt
      181 ttgggatgat tttggttagc ttgagttatc cagttgatcc agacatgata agatacattg
      241 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
      301 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt gatctcccga
      361 tccgtcgacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
      421 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
      481 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
      541 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
      601 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
      661 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
      721 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
      781 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
      841 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
      901 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
      961 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     1021 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     1081 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     1141 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     1201 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     1261 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     1321 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     1381 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     1441 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     1501 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     1561 ctgctgcttg caaacaaaaa aaccgctacc agcggtggtt tgtttgccgg atcaagagct
     1621 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     1681 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     1741 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     1801 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     1861 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     1921 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     1981 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     2041 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     2101 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     2161 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     2221 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     2281 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     2341 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     2401 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     2461 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     2521 ccatgattac gccaagctct agctagaggt cgacggtaaa acgctagcaa tgtcaaggcc
     2581 tctcactctc tgatattcat ttctttgcaa gttataaata ctgaataata agatgacatg
     2641 aactactact gctagagatt ttccacactg actaaaaggg tctgagggat ctctagttac
     2701 cagagtcaca caacagacgg gcacacacta cttgaagcac tcaaggcaag ctttattgag
     2761 gcttaagcag tgggttccct agttagccag agagctccca ggctcagatc tggtctaacc
     2821 agagagaccc agtacaggca aaaagcagct gcttatatgc aggatctgag ggctcgccac
     2881 tccccagtcc cgcccaggcc acgcctccct ggaaagtccc cagcggaaag tcccttgtag
     2941 caagctcgat atcagcagtt cttgaagtac tccggatgca gctctcgggc cacgtgatga
     3001 aatgctaggc ggctgtcaaa cctccactct aacacttctc tctccgggtc atccatccca
     3061 tgcaggctca cagggtgtaa caagctggtg ttctctcctt tattggcctc ttctacctta
     3121 tctggctcaa ctggtactag cttgtagcac catccaaagg tcagtggata tctgacccct
     3181 ggccctggtg tgtagttctg ctaatcaggg aagtagcctt gtgtgtggta gatccacaga
     3241 tcaaggatat cttgtcttcg ttgggagtga attagccctt ccagtcccct cttttctttt
     3301 aaaaagtggc taagatctac agctgccttg taagtcattg gtcttaaagg tacctgaggt
     3361 gtgactggaa aacccacctc ctcctcctct tgtgcttcta gccaggcaca atcagcattg
     3421 gtagctgctg tattgctact tgtgattgct ccatgttttt ctaggtctcg actgcagaat
     3481 taattccagg cggggaggcg gcccaaaggg agatccgact cgtctgaggg cgaaggcgaa
     3541 gacgcggaag aggccgcaga gccggcagca ggccgcggga aggaaggtcc gctggattga
     3601 gggccgaagg gacgtagcag aaggacgtcc cgcgcagaat ccaggtggca acacaggcga
     3661 gcagccatgg aaaggacgtc agcttccccg acaacaccac ggaattgtca gtgcccaaca
     3721 gccgagcccc tgtccagcag cgggcaaggc aggcggcgat gagttccgcc gtggcaatag
     3781 ggagggggaa agcgaaagtc ccggaaagga gctgacaggt ggtggcaatg ccccaaccag
     3841 tgggggttgc gtcagcaaac acagtgcaca ccacgccacg ttgcctgaca acgggccaca
     3901 actcctcata aagagacagc aaccaggatt tatacaagga ggagaaaatg aaagccatac
     3961 gggaagcaat agcatgatac aaaggcatta aagcagcgta tccacatagc gtaaaaggag
     4021 caacatagtt aagaatacca gtcaatcttt cacaaatttt gtaatccaga ggttgattgt
     4081 tccagacgcg ttcaggcacc gggcttgcgg gtcatgcacc aggtgcgcgg tccttcgggc
     4141 acctcgacgt cggcggtgac ggtgaagccg agccgctcgt agaaggggag gttgcggggc
     4201 gcggaggtct ccaggaaggc gggcaccccg gcgcgctcgg ccgcctccac tccggggagc
     4261 acgacggcgc tgcccagacc cttgccctgg tggtcgggcg agacgccgac ggtggccagg
     4321 aaccacgcgg gctccttggg ccggtgcggc gccaggaggc cttccatctg ttgctgcgcg
     4381 gccagccggg aaccgctcaa ctcggccatg cgcgggccga tctcggcgaa caccgccccc
     4441 gcttcgacgc tctccggcgt ggtccagacc gccaccgcgg cgccgtcgtc cgcgacccac
     4501 accttgccga tgtcgagccc gacgcgcgtg aggaagagtt cttgcagctc ggtgacccgc
     4561 tcgatgtggc ggtccgggtc gacggtgtgg cgcgtggcgg ggtagtcggc gaacgcggcg
     4621 gcgagggtgc gtacggcccg ggggacgtcg tcgcgggtgg cgaggcgcac cgtgggcttg
     4681 tactcggtca tggtcgcgtt ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact
     4741 tctggaatag ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc
     4801 agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta
     4861 ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg
     4921 gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac
     4981 ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca ttcgcgcgtt
     5041 tacccgggga gcatgtcaag gtcaaaatcg tcaagagcgt cagcaggcag catatcaagg
     5101 tcaaagtcgt caagggcatc ggctgggagc atgtctaagt caaaatcgtc aagggcgtcg
     5161 gtcggcccgc cgctttcgca ctttagctgt ttctccaggc cacatatgat tagttccagg
     5221 ccgaaaagga aggcaggttc ggctccctgc cggtcgaaca gctcaattgc ttgtttcaga
     5281 agtgggggca tagaatcggt ggtaggtgtc tctctttcct cttttgctac ttgatgctcc
     5341 tgttcctcca atacgcagcc cagtgtaaag tggcccacgg cggacagagc gtacagtgcg
     5401 ttctccaggg agaagccttg ctgacacagg aacgcgagct gattttccag ggtttcgtac
     5461 tgtttctctg ttgggcgggt gccgagatgc actttagccc cgtcgcgatg tgagaggaga
     5521 gcacagcggt atgacttggc gttgttccgc agaaagtctt gccatgactc gccttccagg
     5581 gggcaggagt gggtatgatg cctgtccagc atctcgattg gcagggcatc gagcagggcc
     5641 cgcttgttct tcacgtgcca gtacagggta ggctgctcaa ctcccagctt ttgagcgagt
     5701 ttccttgtcg tcaggccttc gataccgact ccattgagta attccagagc agagtttatg
     5761 actttgctct tgtccagtct agacatggta attcgatgat ccccctgggg agagaggtcg
     5821 gtgattcggt caacgaggga gccgactgcc gacgtgcgct ccggaggctt gcagaatgcg
     5881 gaacaccgcg cgggcaggaa cagggcccac actaccgccc cacaccccgc ctcccgcacc
     5941 gccccttccc ggccgctgct ctcggcgcgc cctgctgagc agccgctatt ggccacagcc
     6001 catcgcggtc ggcgcgctgc cattgctccc tggcgctgtc cgtctgcgag ggtactagtg
     6061 agacgtgcgg cttccgtttg tcacgtccgg cacgccgcga accgcaagga accttcccga
     6121 cttaggggcg gagcaggaag cgtcgccggg gggcccacaa gggtagcggc gaagatccgg
     6181 gtgacgctgc gaacggacgt gaagaatgtg cgagacccag ggtcggcgcc gctgcgtttc
     6241 ccggaaccac gcccagagca gccgcgtccc tgcgcaaacc cagggctgcc ttggaaaagg
     6301 cgcaacccca accccgtgga attatcacct cgagtttact ccctatcagt gatagagaac
     6361 gtatgaagag tttactccct atcagtgata gagaacgtat gcagacttta ctccctatca
     6421 gtgatagaga acgtataagg agtttactcc ctatcagtga tagagaacgt atgaccagtt
     6481 tactccctat cagtgataga gaacgtatct acagtttact ccctatcagt gatagagaac
     6541 gtatatccag tttactccct atcagtgata gagaacgtat aagctttgct tatgtaaacc
     6601 agggcgccta taaaagagtg ctgatttttt gagtaaactt caattccaca acacttttgt
     6661 cttataccaa ctttccgtac cacttcctac cctcgtaaag aattcccgta tacaccggtg
     6721 ccggcggatc cgactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca
     6781 aggatgacga tgacaagttc gaacaccatc accatcacca tcaccatcac catcatcact
     6841 agagaccacc tcccctgcga gctaagctgg acagccaatg acgggtaaga gagtgacatt
     6901 tttcactaac ctaagacagg agggccgtca gagctactgc ctaatccaaa gacgggtaaa
     6961 agtgataaaa atgtatcact ccaacctaag acaggcgcag cttccgaggg atttgagatc
     7021 cagacatgat aagatacatt gatgagtttg gacaaaccaa aactagaatg cagtgaaaaa
     7081 aatgccttat ttgtgaaatt tgtgatgcta ttgccttatt tgtaaccatt ataagctgca
     7141 ataaacaagt taagtcgata aactggatct ctgctgtccc tgtaataaac ccgaaaattt
     7201 tgaatttttg taatttgttt ttgtagttct ttagtttgta tgtctgttgc tattatgtct
     7261 actattcttt cccctgcact gtacccccca atcccccctt ttcttttaaa ggcgataccg
     7321 tcgagatccg ttcactaatc gaatggatct gtctctgtct ctctctccac cttcttcttc
     7381 tattccttcg ggcctgtcgg gtcccctcgg ggttgggagg tgggtctgaa acgataatgg
     7441 tgaatatccc tgcctaactc tattcactat agaaagtaca gcaaaaacta ttcttaaacc
     7501 taccaagcct cctactatca ttatgaataa ttttatatac cacagccaat ttgttatgtt
     7561 aaaccaattc cacaaacttg cccatttatc taattccaat aattcttgtt cattcttttc
     7621 ttgctggttt tgcgattctt caattaagga gtgtattaag cttgtgtaat tgttaatttc
     7681 tctgtcccac tccatccagg tcgtgtgatt ccaaatctgt tccagagatt tattactcca
     7741 actagcattc caaggcacag cagtggtgca aatgagtttt ccagagcaac cccaaatccc
     7801 caggagctgt tgatccttta ggtatctttc cacagccagg attcttgcct ggagctgctt
     7861 gatgccccag actgtgagtt gcaacagatg ctgttgcgcc tcaatagccc tcagcaaatt
     7921 gttctgctgc tgcactatac cagacaataa ttgtctggcc tgtaccgtca gcgtcattga
     7981 cgctgcgccc atagtgcttc ctgctgctcc caagaaccca aggaacaaag ctcctattcc
     8041 cactgctctt ttttctctct gcaccactct tctctttgcc ttggtgggtg ctactcctaa
     8101 tggttcaatt tttactactt tatatttata taattcactt ctccaattgt ccctcatatc
     8161 tcctcctcca ggtctgaaga tcagcggccg gccgcttgct gtgcggtggt cttacttttg
     8221 ttttgctctt cctctatctt gtctaaagct tccttggtgt cttttatctc tatcctttga
     8281 tgcacacaat agagggttgc tactgtatta tataatgatc taagttcttc tgatcctgtc
     8341 tgaagggatg gttgtagctg tcccagtatt tgtctacagc cttctgatgt ttctaacagg
     8401 ccaggattaa ctgcgaatcg ttctagctcc ctgcttgccc atactatatg ttttaattta
     8461 tattttttct ttccccctgg ccttaaccga attttttccc atcgcgatct aattctcccc
     8521 cgcttaatac tgacgctctc gcacccatct ctctccttct agcctccgct agtcaaaatt
     8581 tttggcgtac tcaccagtcg ccgcccctcg cctcttgccg tgcgcgcttc agcaagccga
     8641 gtcctgcgtc gagagagctc ctctggtttc cctttcgctt tcaagtccct gttcgggcgc
     8701 cactgctaga gattttccac actgactaaa agggtctgag ggatctctag ttaccagagt
     8761 cacacaacag acgggcacac actacttgaa gcactcaagg caagctttat tgaggcttaa
     8821 gcagtgggtt ccctagttag ccagagagct cccaggctca gatctggtct aaccagagag
     8881 acccagtaca ggcaaaaagc agctgcttat atgcaggatc tgagggctcg ccactcccca
     8941 gtcccgccca ggccacgcct ccctggaaag tccccagcgg aaagtccctt gtagcaagct
     9001 cgatatcagc agttcttgaa gtactccgga tgcagctctc gggccacgtg atgaaatgct
     9061 aggcggctgt caaacctcca ctctaacact tctctctccg ggtcatccat cccatgcagg
     9121 ctcacagggt gtaacaagct ggtgttctct cctttattgg cctcttctac cttatctggc
     9181 tcaactggta ctagcttgta gcaccatcca aaggtcagtg gatatctgac ccctggccct
     9241 ggtgtgtagt tctgctaatc agggaagtag ccttgtgtgt ggtagatcca cagatcaagg
     9301 atatcttgtc ttctttggga gtgaattagc ccttcca
//
