LOCUS       .                       9735 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(4094..4489)
                     /label="BSD"
                     /note="blasticidin S deaminase"
     promoter        complement(4504..4833)
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             complement(4847..5590)
                     /label="Tet-On® 3G"
                     /note="modified rtTA protein that binds tightly to
                     promoters containing the tet operator in the presence of
                     doxycycline"
     promoter        complement(5609..6119)
                     /label="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
     promoter        6136..6503
                     /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             6524..7231
                     /label="mCherry"
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     misc_feature    complement(7592..7709)
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     CDS             complement(7781..7822)
                     /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(7971..8015)
                     /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(8200..8433)
                     /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(8930..9055)
                     /label="HIV-1 Ψ"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     LTR             complement(9102..9735)
                     /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 ttagccctcc cacacataac cagagggcag caattcacga atcccaactg
     4141 ccgtcggctg tccatcactg tccttcacta tggctttgat cccaggatgc agatcgagaa
     4201 gcacctgtcg gcaccgtccg caggggctca agatgcccct gttctcattt ccgatcgcga
     4261 cgatacaagt caggttgcca gctgccgcag cagcagcagt gcccagcacc acgagttctg
     4321 cacaaggtcc cccagtaaaa tgatatacat tgacaccagt gaagatgcgg ccgtcgctag
     4381 agagagctgc gctggcgacg ctgtagtctt cagagatggg gatgctgttg attgtagccg
     4441 ttgctctttc aatgagggtg gattcttctt gagacaaagg cttggccatg gtggcggtcg
     4501 cgttttgcaa aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga
     4561 ggccgaggcg gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga
     4621 atgggcggaa ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt
     4681 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     4741 tccacacctg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga
     4801 gcctggggac tttccacacc ctaactgaca cacattcgcg cgtttacccg gggagcatgt
     4861 caaggtcaaa atcgtcaaga gcgtcagcag gcagcatatc aaggtcaaag tcgtcaaggg
     4921 catcggctgg gagcatgtct aagtcaaaat cgtcaagggc gtcggtcggc ccgccgcttt
     4981 cgcactttag ctgtttctcc aggccacata tgattagttc caggccgaaa aggaaggcag
     5041 gttcggctcc ctgccggtcg aacagctcaa ttgcttgttt cagaagtggg ggcatagaat
     5101 cggtggtagg tgtctctctt tcctcttttg ctacttgatg ctcctgttcc tccaatacgc
     5161 agcccagtgt aaagtggccc acggcggaca gagcgtacag tgcgttctcc agggagaagc
     5221 cttgctgaca caggaacgcg agctgatttt ccagggtttc gtactgtttc tctgttgggc
     5281 gggtgccgag atgcacttta gccccgtcgc gatgtgagag gagagcacag cggtatgact
     5341 tggcgttgtt ccgcagaaag tcttgccatg actcgccttc cagggggcag gagtgggtat
     5401 gatgcctgtc cagcatctcg attggcaggg catcgagcag ggcccgcttg ttcttcacgt
     5461 gccagtacag ggtaggctgc tcaactccca gcttttgagc gagtttcctt gtcgtcaggc
     5521 cttcgatacc gactccattg agtaattcca gagcagagtt tatgactttg ctcttgtcca
     5581 gtctagacat ggtaattcga tgatccccct ggggagagag gtcggtgatt cggtcaacga
     5641 gggagccgac tgccgacgtg cgctccggag gcttgcagaa tgcggaacac cgcgcgggca
     5701 ggaacagggc ccacactacc gccccacacc ccgcctcccg caccgcccct tcccggccgc
     5761 tgctctcggc gcgccctgct gagcagccgc tattggccac agcccatcgc ggtcggcgcg
     5821 ctgccattgc tccctggcgc tgtccgtctg cgagggtact agtgagacgt gcggcttccg
     5881 tttgtcacgt ccggcacgcc gcgaaccgca aggaaccttc ccgacttagg ggcggagcag
     5941 gaagcgtcgc cggggggccc acaagggtag cggcgaagat ccgggtgacg ctgcgaacgg
     6001 acgtgaagaa tgtgcgagac ccagggtcgg cgccgctgcg tttcccggaa ccacgcccag
     6061 agcagccgcg tccctgcgca aacccagggc tgccttggaa aaggcgcaac cccaaccccg
     6121 tggaattatc acctcgagtt tactccctat cagtgataga gaacgtatga agagtttact
     6181 ccctatcagt gatagagaac gtatgcagac tttactccct atcagtgata gagaacgtat
     6241 aaggagttta ctccctatca gtgatagaga acgtatgacc agtttactcc ctatcagtga
     6301 tagagaacgt atctacagtt tactccctat cagtgataga gaacgtatat ccagtttact
     6361 ccctatcagt gatagagaac gtataagctt tgcttatgta aaccagggcg cctataaaag
     6421 agtgctgatt ttttgagtaa acttcaattc cacaacactt ttgtcttata ccaactttcc
     6481 gtaccacttc ctaccctcgt aaagaattcc cgtatacacc ggtatggtga gcaagggcga
     6541 ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc
     6601 cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
     6661 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct
     6721 gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga
     6781 ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
     6841 cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa
     6901 ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat
     6961 gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat
     7021 caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta
     7081 caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat
     7141 cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca
     7201 ctccaccggc ggcatggacg agctgtacaa gtagggatcc agaccacctc ccctgcgagc
     7261 taagctggac agccaatgac gggtaagaga gtgacatttt tcactaacct aagacaggag
     7321 ggccgtcaga gctactgcct aatccaaaga cgggtaaaag tgataaaaat gtatcactcc
     7381 aacctaagac aggcgcagct tccgagggat ttgagatcca gacatgataa gatacattga
     7441 tgagtttgga caaaccaaaa ctagaatgca gtgaaaaaaa tgccttattt gtgaaatttg
     7501 tgatgctatt gccttatttg taaccattat aagctgcaat aaacaagtta agtcgataaa
     7561 ctggatctct gctgtccctg taataaaccc gaaaattttg aatttttgta atttgttttt
     7621 gtagttcttt agtttgtatg tctgttgcta ttatgtctac tattctttcc cctgcactgt
     7681 accccccaat cccccctttt cttttaaagg cgataccgtc gagatccgtt cactaatcga
     7741 atggatctgt ctctgtctct ctctccacct tcttcttcta ttccttcggg cctgtcgggt
     7801 cccctcgggg ttgggaggtg ggtctgaaac gataatggtg aatatccctg cctaactcta
     7861 ttcactatag aaagtacagc aaaaactatt cttaaaccta ccaagcctcc tactatcatt
     7921 atgaataatt ttatatacca cagccaattt gttatgttaa accaattcca caaacttgcc
     7981 catttatcta attccaataa ttcttgttca ttcttttctt gctggttttg cgattcttca
     8041 attaaggagt gtattaagct tgtgtaattg ttaatttctc tgtcccactc catccaggtc
     8101 gtgtgattcc aaatctgttc cagagattta ttactccaac tagcattcca aggcacagca
     8161 gtggtgcaaa tgagttttcc agagcaaccc caaatcccca ggagctgttg atcctttagg
     8221 tatctttcca cagccaggat tcttgcctgg agctgcttga tgccccagac tgtgagttgc
     8281 aacagatgct gttgcgcctc aatagccctc agcaaattgt tctgctgctg cactatacca
     8341 gacaataatt gtctggcctg taccgtcagc gtcattgacg ctgcgcccat agtgcttcct
     8401 gctgctccca agaacccaag gaacaaagct cctattccca ctgctctttt ttctctctgc
     8461 accactcttc tctttgcctt ggtgggtgct actcctaatg gttcaatttt tactacttta
     8521 tatttatata attcacttct ccaattgtcc ctcatatctc ctcctccagg tctgaagatc
     8581 agcggccggc cgcttgctgt gcggtggtct tacttttgtt ttgctcttcc tctatcttgt
     8641 ctaaagcttc cttggtgtct tttatctcta tcctttgatg cacacaatag agggttgcta
     8701 ctgtattata taatgatcta agttcttctg atcctgtctg aagggatggt tgtagctgtc
     8761 ccagtatttg tctacagcct tctgatgttt ctaacaggcc aggattaact gcgaatcgtt
     8821 ctagctccct gcttgcccat actatatgtt ttaatttata ttttttcttt ccccctggcc
     8881 ttaaccgaat tttttcccat cgcgatctaa ttctcccccg cttaatactg acgctctcgc
     8941 acccatctct ctccttctag cctccgctag tcaaaatttt tggcgtactc accagtcgcc
     9001 gcccctcgcc tcttgccgtg cgcgcttcag caagccgagt cctgcgtcga gagagctcct
     9061 ctggtttccc tttcgctttc aagtccctgt tcgggcgcca ctgctagaga ttttccacac
     9121 tgactaaaag ggtctgaggg atctctagtt accagagtca cacaacagac gggcacacac
     9181 tacttgaagc actcaaggca agctttattg aggcttaagc agtgggttcc ctagttagcc
     9241 agagagctcc caggctcaga tctggtctaa ccagagagac ccagtacagg caaaaagcag
     9301 ctgcttatat gcaggatctg agggctcgcc actccccagt cccgcccagg ccacgcctcc
     9361 ctggaaagtc cccagcggaa agtcccttgt agcaagctcg atatcagcag ttcttgaagt
     9421 actccggatg cagctctcgg gccacgtgat gaaatgctag gcggctgtca aacctccact
     9481 ctaacacttc tctctccggg tcatccatcc catgcaggct cacagggtgt aacaagctgg
     9541 tgttctctcc tttattggcc tcttctacct tatctggctc aactggtact agcttgtagc
     9601 accatccaaa ggtcagtgga tatctgaccc ctggccctgg tgtgtagttc tgctaatcag
     9661 ggaagtagcc ttgtgtgtgg tagatccaca gatcaaggat atcttgtctt ctttgggagt
     9721 gaattagccc ttcca
//
