LOCUS       .                      10220 bp    DNA     linear   UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     protein_bind    113..134
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        149..179
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     promoter        248..266
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        294..520
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             521..701
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    748..873
                     /label="HIV-1 Ψ"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1366..1599
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             1784..1828
                     /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             1977..2018
                     /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    2095..2211
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     enhancer        2234..2537
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2538..2741
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             2905..3612
                     /label="mCherry"
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     promoter        3646..3975
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             3985..5634
                     /label="luciferase"
                     /note="firefly luciferase"
     CDS             5704..5757
                     /label="T2A"
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             5758..6354
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    6358..6945
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             7032..7265
                     /label="3' LTR (ΔU3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7337..7471
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     promoter        complement(7654..7672)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      7840..8295
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        8321..8425
                     /label="AmpR promoter"
     CDS             8426..9283
                     /label="AmpR"
                     /note="β-lactamase"
ORIGIN
        1 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
       61 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
      121 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
      181 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
      241 agcgcgcaat taaccctcac taaagggaac aaaagctgga gctgcaagct taatgtagtc
      301 ttatgcaata ctcttgtagt cttgcaacat ggtaacgatg agttagcaac atgccttaca
      361 aggagagaaa aagcaccgtg catgccgatt ggtggaagta aggtggtacg atcgtgcctt
      421 attaggaagg caacagacgg gtctgacatg gattggacga accactgaat tgccgcattg
      481 cagagatatt gtatttaagt gcctagctcg atacataaac gggtctctct ggttagacca
      541 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
      601 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
      661 atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac
      721 ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg
      781 cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga
      841 ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg
      901 cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata
      961 gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca
     1021 gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa
     1081 cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata
     1141 aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc
     1201 gcacagcaag cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag
     1261 aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa
     1321 ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct
     1381 tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca
     1441 ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga
     1501 ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat
     1561 cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg
     1621 aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga
     1681 acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag
     1741 cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt
     1801 attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg
     1861 gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc
     1921 tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca
     1981 cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag gtggagagag
     2041 agacagagac agatccattc gattagtgaa cggatctcga cggtatcggt taacttttaa
     2101 aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac
     2161 agacatacaa actaaagaat tacaaaaaca aattacaaaa ttcaaaattt tatcgataag
     2221 cttgggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     2281 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     2341 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     2401 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     2461 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     2521 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     2581 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     2641 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     2701 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga
     2761 tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccga ctctagacgg
     2821 gatccggcta gcgactagtt actcgaggct actaacttca gcctgctgaa gcaggctgga
     2881 gacgtggagg agaaccctgg acctatggtg agcaagggcg aggaggataa catggccatc
     2941 atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc
     3001 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag
     3061 gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac
     3121 ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc
     3181 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg
     3241 acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc
     3301 aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc
     3361 gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg
     3421 aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg
     3481 cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac
     3541 tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac
     3601 gagctgtaca agtaacgcgt cgaccagtgg tcgaccctgt ggaatgtgtg tcagttaggg
     3661 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3721 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3781 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3841 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3901 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3961 ctaggctttt gcaaaaagct taccatggaa gacgccaaaa acataaagaa aggcccggcg
     4021 ccattctatc cgctggaaga tggaaccgct ggagagcaac tgcataaggc tatgaagaga
     4081 tacgccctgg ttcctggaac aattgctttt acagatgcac atatcgaggt ggacatcact
     4141 tacgctgagt acttcgaaat gtccgttcgg ttggcagaag ctatgaaacg atatgggctg
     4201 aatacaaatc acagaatcgt cgtatgcagt gaaaactctc ttcaattctt tatgccggtg
     4261 ttgggcgcgt tatttatcgg agttgcagtt gcgcccgcga acgacattta taatgaacgt
     4321 gaattgctca acagtatggg catttcgcag cctaccgtgg tgttcgtttc caaaaagggg
     4381 ttgcaaaaaa ttttgaacgt gcaaaaaaag ctcccaatca tccaaaaaat tattatcatg
     4441 gattctaaaa cggattacca gggatttcag tcgatgtaca cgttcgtcac atctcatcta
     4501 cctcccggtt ttaatgaata cgattttgtg ccagagtcct tcgataggga caagacaatt
     4561 gcactgatca tgaactcctc tggatctact ggtctgccta aaggtgtcgc tctgcctcat
     4621 agaactgcct gcgtgagatt ctcgcatgcc agagatccta tttttggcaa tcaaatcatt
     4681 ccggatactg cgattttaag tgttgttcca ttccatcacg gttttggaat gtttactaca
     4741 ctcggatatt tgatatgtgg atttcgagtc gtcttaatgt atagatttga agaagagctg
     4801 tttctgagga gccttcagga ttacaagatt caaagtgcgc tgctggtgcc aaccctattc
     4861 tccttcttcg ccaaaagcac tctgattgac aaatacgatt tatctaattt acacgaaatt
     4921 gcttctggtg gcgctcccct ctctaaggaa gtcggggaag cggttgccaa gaggttccat
     4981 ctgccaggta tcaggcaagg atatgggctc actgagacta catcagctat tctgattaca
     5041 cccgaggggg atgataaacc gggcgcggtc ggtaaagttg ttccattttt tgaagcgaag
     5101 gttgtggatc tggataccgg gaaaacgctg ggcgttaatc aaagaggcga actgtgtgtg
     5161 agaggtccta tgattatgtc cggttatgta aacaatccgg aagcgaccaa cgccttgatt
     5221 gacaaggatg gatggctaca ttctggagac atagcttact gggacgaaga cgaacacttc
     5281 ttcatcgttg accgcctgaa gtctctgatt aagtacaaag gctatcaggt ggctcccgct
     5341 gaattggaat ccatcttgct ccaacacccc aacatcttcg acgcaggtgt cgcaggtctt
     5401 cccgacgatg acgccggtga acttcccgcc gccgttgttg ttttggagca cggaaagacg
     5461 atgacggaaa aagagatcgt ggattacgtc gccagtcaag taacaaccgc gaaaaagttg
     5521 cgcggaggag ttgtgtttgt ggacgaagta ccgaaaggtc ttaccggaaa actcgacgca
     5581 agaaaaatca gagagatcct cataaaggcc aagaagggcg gaaagatcgc cgtgagatcc
     5641 cgcgctcagt cgtccaattc tgccgtggac ggcaccgccg gacccggctc caccggatct
     5701 cgcgagggca gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc cggccctatg
     5761 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc
     5821 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc
     5881 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc
     5941 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc
     6001 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc
     6061 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc
     6121 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc
     6181 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag
     6241 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac
     6301 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgaaat
     6361 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     6421 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     6481 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     6541 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     6601 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     6661 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     6721 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc
     6781 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     6841 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtctccgc
     6901 cttcgccctc agacgagtcg gatctccctt tggccgcctc cccgcctgga attcgagctc
     6961 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     7021 aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg
     7081 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     7141 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     7201 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     7261 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat
     7321 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     7381 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     7441 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc
     7501 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     7561 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     7621 ttttttggag gcctagggac gtacccaatt cgccctatag tgagtcgtat tacgcgcgct
     7681 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     7741 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     7801 gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
     7861 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     7921 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     7981 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     8041 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     8101 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     8161 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     8221 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     8281 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     8341 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     8401 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     8461 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     8521 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     8581 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     8641 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     8701 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     8761 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     8821 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     8881 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     8941 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     9001 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     9061 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     9121 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     9181 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     9241 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     9301 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     9361 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     9421 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     9481 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     9541 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     9601 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     9661 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     9721 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     9781 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     9841 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     9901 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     9961 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
    10021 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
    10081 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
    10141 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
    10201 cgagtcagtg agcgaggaag
//
