LOCUS       .                       8445 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        65..368
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        369..572
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     protein_bind    595..619
                     /label="attB1"
                     /note="recombination site for the Gateway® BP reaction"
     CDS             769..1089
                     /label="FKBP"
                     /note="human FK506-binding protein FKBP12"
     CDS             1120..1161
                     /label="V5 tag"
                     /note="epitope tag from simian virus 5"
     misc_feature    1452..2040
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             2112..2345
                     /label="3' LTR (ΔU3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2423..2557
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      2584..2719
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(2740..2758)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2768..2784)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      2926..3381
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3407..3511
                     /label="AmpR promoter"
     CDS             3512..4369
                     /label="AmpR"
                     /note="β-lactamase"
     rep_origin      4543..5131
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    5419..5440
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5455..5485
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    5493..5509
                     /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     5517..5533
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        5554..5572
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5600..5826
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     misc_feature    6054..6179
                     /label="HIV-1 Ψ"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    6672..6905
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             7090..7134
                     /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        7309..7809
                     /label="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             7821..8216
                     /label="BSD"
                     /note="blasticidin S deaminase"
     misc_feature    8276..8393
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
ORIGIN
        1 cccggggtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
       61 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
      121 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      181 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      241 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      301 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      361 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      421 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      481 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      541 gtgtacggtg ggaggtctat ataagcagag ctctctggct aactgtcggg atcaacaagt
      601 ttgtacaaaa aagcaggctt cgaaggagat agaaccatgg tgggtcggaa cagcgccatc
      661 gccgcgggcg tgtgcggtgc cctcttcata gggtactgca tctactttga ccgcaaaaga
      721 cgaagtgacc ccaacttcgg atccggagca agtggaagtg ctggtggtgg agtgcaggtg
      781 gaaaccatct ccccaggaga cgggcgcacc ttccccaagc gcggccagac ctgcgtggtg
      841 cactacaccg ggatgcttga agatggaaag aaatttgatt cctcccggga cagaaacaag
      901 ccctttaagt ttatgctagg caagcaggag gtgatccgag gctgggaaga aggggttgcc
      961 cagatgagtg tgggtcagag agccaaactg actatatctc cagattatgc ctatggtgcc
     1021 actgggcacc caggcatcat cccaccacat gccactctcg tcttcgatgt ggagcttcta
     1081 aaactggaag ggagtggcag cggaggtagt gggagcggtg gcaagcccat ccccaacccc
     1141 ctgctgggcc tggacagcac caagggctcg ggctcgacct cgggctcggg caccggtaaa
     1201 gacaatactg tgcctctgaa gctgatcgct ctcctggcta atggcgagtt ccatagtggc
     1261 gaacagctgg gagaaaccct gggcatgtcc agggccgcta tcaacaagca cattcagact
     1321 ctgcgcgact ggggcgtgga cgtgttcacc gtgcccggaa agggctactc tctgcccgag
     1381 cctatcccgc tgctgaacgc taaacagatt ctgtaataag ctagcgctaa ccggtggcgc
     1441 gttaagtcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     1501 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     1561 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     1621 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     1681 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     1741 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     1801 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     1861 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     1921 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     1981 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     2041 gtcgacttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     2101 aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg
     2161 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     2221 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     2281 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     2341 tagcagtacg tatagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa
     2401 atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag
     2461 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     2521 gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg
     2581 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     2641 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     2701 ggaggctttt ttggaggcct agggacgtac ccaattcgcc ctatagtgag tcgtattacg
     2761 cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     2821 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     2881 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg
     2941 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     3001 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     3061 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     3121 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     3181 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     3241 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     3301 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     3361 aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat
     3421 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     3481 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     3541 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     3601 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     3661 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     3721 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     3781 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     3841 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     3901 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     3961 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     4021 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     4081 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     4141 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     4201 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     4261 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     4321 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     4381 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     4441 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     4501 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     4561 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     4621 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     4681 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     4741 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     4801 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     4861 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     4921 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     4981 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     5041 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5101 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5161 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     5221 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     5281 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     5341 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     5401 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     5461 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     5521 aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa
     5581 gctggagctg caagcttaat gtagtcttat gcaatactct tgtagtcttg caacatggta
     5641 acgatgagtt agcaacatgc cttacaagga gagaaaaagc accgtgcatg ccgattggtg
     5701 gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac agacgggtct gacatggatt
     5761 ggacgaacca ctgaattgcc gcattgcaga gatattgtat ttaagtgcct agctcgatac
     5821 ataaacgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     5881 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     5941 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     6001 tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc
     6061 gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga
     6121 gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca
     6181 gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa
     6241 agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag
     6301 ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac
     6361 catcccttca gacaggatca gaagaactta gatcattata taatacagta gcaaccctct
     6421 attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg
     6481 aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt cagacctgga
     6541 ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt
     6601 gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga
     6661 gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc
     6721 gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag
     6781 cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg
     6841 ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag
     6901 ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat
     6961 gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg
     7021 gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac
     7081 cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat
     7141 tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc
     7201 ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga
     7261 tattcaccat tatcgtttca gacccacctc ccaaccccga ggggaccctt gcgccttttc
     7321 caaggcagcc ctgggtttgc gcagggacgc ggctgctctg ggcgtggttc cgggaaacgc
     7381 agcggcgccg accctgggtc tcgcacattc ttcacgtccg ttcgcagcgt cacccggatc
     7441 ttcgccgcta cccttgtggg ccccccggcg acgcttcctg ctccgcccct aagtcgggaa
     7501 ggttccttgc ggttcgcggc gtgccggacg tgacaaacgg aagccgcacg tctcactagt
     7561 accctcgcag acggacagcg ccagggagca atggcagcgc gccgaccgcg atgggctgtg
     7621 gccaatagcg gctgctcagc agggcgcgcc gagagcagcg gccgggaagg ggcggtgcgg
     7681 gaggcggggt gtggggcggt agtgtgggcc ctgttcctgc ccgcgcggtg ttccgcattc
     7741 tgcaagcctc cggagcgcac gtcggcagtc ggctccctcg ttgaccgaat caccgacctc
     7801 tctccccagg gggtaccacc atggccaagc ctttgtctca agaagaatcc accctcattg
     7861 aaagagcaac ggctacaatc aacagcatcc ccatctctga agactacagc gtcgccagcg
     7921 cagctctctc tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat tttactgggg
     7981 gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc tgcggcagct ggcaacctga
     8041 cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt gagcccctgc ggacggtgcc
     8101 gacaggtgct tctcgatctg catcctggga tcaaagccat agtgaaggac agtgatggac
     8161 agccgacggc agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg gagggcctgc
     8221 agctgcagta gtaagaattc tagatcttga gacaaatggc agtattcatc cacaatttta
     8281 aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa
     8341 cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt
     8401 attacaggga cagcagagat ccactttggc gccggctcga ggggg
//
