LOCUS       .                       7824 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     protein_bind    135..156
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        171..201
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    209..225
                     /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     233..249
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        270..288
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        316..542
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     misc_feature    770..895
                     /label="HIV-1 Ψ"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1388..1621
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             1806..1850
                     /label="gp41 peptide"
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
                     al., 2013)"
     promoter        2025..2525
                     /label="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             2537..2932
                     /label="BSD"
                     /note="blasticidin S deaminase"
     misc_feature    2992..3109
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     enhancer        3226..3529
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3530..3733
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     protein_bind    3756..3780
                     /label="attB1"
                     /note="recombination site for the Gateway® BP reaction"
     CDS             3801..3824
                     /label="FLAG"
                     /note="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             3849..3875
                     /label="NES"
                     /note="nuclear export signal from the HIV Rev protein
                     (Fischer et al., 1995)"
     protein_bind    complement(3883..3907)
                     /label="attB2"
                     /note="recombination site for the Gateway® BP reaction"
     CDS             3909..3950
                     /label="V5 tag"
                     /note="epitope tag from simian virus 5"
     misc_feature    3992..4580
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             4652..4885
                     /label="3' LTR (ΔU3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4963..5097
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      5124..5259
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(5280..5298)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5308..5324)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      5466..5921
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        5947..6051
                     /label="AmpR promoter"
     CDS             6052..6909
                     /label="AmpR"
                     /note="β-lactamase"
     rep_origin      7083..7671
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
ORIGIN
        1 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
       61 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
      121 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
      181 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
      241 agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg
      301 gagctgcaag cttaatgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga
      361 tgagttagca acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag
      421 taaggtggta cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac
      481 gaaccactga attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacataa
      541 acgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
      601 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
      661 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
      721 gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg
      781 caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac
      841 gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat
      901 taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa
      961 aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa
     1021 tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc
     1081 ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg
     1141 tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga
     1201 gcaaaacaaa agtaagacca ccgcacagca agcggccgct gatcttcaga cctggaggag
     1261 gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac
     1321 cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag
     1381 tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag
     1441 cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga
     1501 acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca
     1561 tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc
     1621 tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta
     1681 gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca
     1741 gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc
     1801 aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt
     1861 ttaacataac aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg
     1921 taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt
     1981 caccattatc gtttcagacc cacctcccaa ccccgagggg acccttgcgc cttttccaag
     2041 gcagccctgg gtttgcgcag ggacgcggct gctctgggcg tggttccggg aaacgcagcg
     2101 gcgccgaccc tgggtctcgc acattcttca cgtccgttcg cagcgtcacc cggatcttcg
     2161 ccgctaccct tgtgggcccc ccggcgacgc ttcctgctcc gcccctaagt cgggaaggtt
     2221 ccttgcggtt cgcggcgtgc cggacgtgac aaacggaagc cgcacgtctc actagtaccc
     2281 tcgcagacgg acagcgccag ggagcaatgg cagcgcgccg accgcgatgg gctgtggcca
     2341 atagcggctg ctcagcaggg cgcgccgaga gcagcggccg ggaaggggcg gtgcgggagg
     2401 cggggtgtgg ggcggtagtg tgggccctgt tcctgcccgc gcggtgttcc gcattctgca
     2461 agcctccgga gcgcacgtcg gcagtcggct ccctcgttga ccgaatcacc gacctctctc
     2521 cccagggggt accaccatgg ccaagccttt gtctcaagaa gaatccaccc tcattgaaag
     2581 agcaacggct acaatcaaca gcatccccat ctctgaagac tacagcgtcg ccagcgcagc
     2641 tctctctagc gacggccgca tcttcactgg tgtcaatgta tatcatttta ctgggggacc
     2701 ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca acctgacttg
     2761 tatcgtcgcg atcggaaatg agaacagggg catcttgagc ccctgcggac ggtgccgaca
     2821 ggtgcttctc gatctgcatc ctgggatcaa agccatagtg aaggacagtg atggacagcc
     2881 gacggcagtt gggattcgtg aattgctgcc ctctggttat gtgtgggagg gcctgcagct
     2941 gcagtagtaa gaattctaga tcttgagaca aatggcagta ttcatccaca attttaaaag
     3001 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
     3061 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta
     3121 cagggacagc agagatccac tttggcgccg gctcgagggg gcccggggtt attaatagta
     3181 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     3241 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     3301 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     3361 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     3421 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     3481 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     3541 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     3601 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     3661 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     3721 tataagcaga gctctctggc taactgtcgg gatcaacaag tttgtacaaa aaagcaggct
     3781 tcgaaggaga tagaaccatg gactacaagg atgacgacga taagggatcc acttacgata
     3841 tcctgcagct gcctcccctg gagcgcctga ccctggacta agacccagct ttcttgtaca
     3901 aagtggttgg taagcctatc cctaaccctc tcctcggtct cgattctacg tagtaatgag
     3961 ctagcgctaa ccggtggcgc gttaagtcga caatcaacct ctggattaca aaatttgtga
     4021 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     4081 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     4141 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     4201 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     4261 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     4321 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     4381 ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     4441 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     4501 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     4561 cctttgggcc gcctccccgc gtcgacttta agaccaatga cttacaaggc agctgtagat
     4621 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga
     4681 caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag
     4741 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     4801 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     4861 tagtcagtgt ggaaaatctc tagcagtacg tatagtagtt catgtcatct tattattcag
     4921 tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct
     4981 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     5041 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag
     5101 ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     5161 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     5221 agctattcca gaagtagtga ggaggctttt ttggaggcct agggacgtac ccaattcgcc
     5281 ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga
     5341 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     5401 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     5461 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     5521 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     5581 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     5641 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     5701 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     5761 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     5821 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     5881 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga
     5941 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     6001 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     6061 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     6121 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     6181 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     6241 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     6301 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     6361 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     6421 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     6481 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     6541 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     6601 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     6661 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     6721 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     6781 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     6841 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     6901 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     6961 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     7021 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     7081 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     7141 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     7201 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     7261 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     7321 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     7381 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     7441 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     7501 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     7561 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     7621 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     7681 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     7741 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     7801 cgccgcagcc gaacgaccga gcgc
//
