LOCUS       V035740                 9329 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035740
VERSION     V035740
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             1..634
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     misc_feature    681..806
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1303..1536
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1721..1765
                     /codon_start=1
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /product="antigenic peptide corresponding to amino acids
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
                     et al., 2013)"
                     /transl_table=1
                     /translation="KNEQELLELDKWASL"
                     /label="gp41 peptide"
     misc_feature    2028..2143
                     /note="central polypurine tract and central termination
                     sequence of HIV-1 (lacking the first T)"
                     /label="cPPT/CTS"
     enhancer        2201..2504
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2505..2708
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2829..2855
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2856..4085
                     /label="CCNE1(NM_001238)"
                     /note="CCNE1(NM_001238)"
                     /gene="CCNE1"
     promoter        4117..4616
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4637..5236
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     misc_feature    5250..5838
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6045..6678
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6807..6823)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6831..6847
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
                     /label="lac operator"
     promoter        complement(6855..6885)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6900..6921
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7209..7797)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7968..8828)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(8829..8933)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    8981..9115
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa
     2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     2761 tccatagaag acaccgactc tactagagga tcgctagcgc taccggactc agatctcgag
     2821 ccaccatgta cccatacgac gtaccagatt acgctatgcc gagggagcgc agggagcggg
     2881 atgcgaagga gcgggacacc atgaaggagg acggcggcgc ggagttctcg gctcgctcca
     2941 ggaagaggaa ggcaaacgtg accgtttttt tgcaggatcc agatgaagaa atggccaaaa
     3001 tcgacaggac ggcgagggac cagtgtggga gccagccttg ggacaataat gcagtctgtg
     3061 cagacccctg ctccctgatc cccacacctg acaaagaaga tgatgaccgg gtttacccaa
     3121 actcaacgtg caagcctcgg attattgcac catccagagg ctccccgctg cctgtactga
     3181 gctgggcaaa tagagaggaa gtctggaaaa tcatgttaaa caaggaaaag acatacttaa
     3241 gggatcagca ctttcttgag caacaccctc ttctgcagcc aaaaatgcga gcaattcttc
     3301 tggattggtt aatggaggtg tgtgaagtct ataaacttca cagggagacc ttttacttgg
     3361 cacaagattt ctttgaccgg tatatggcga cacaagaaaa tgttgtaaaa actcttttac
     3421 agcttattgg gatttcatct ttatttattg cagccaaact tgaggaaatc tatcctccaa
     3481 agttgcacca gtttgcgtat gtgacagatg gagcttgttc aggagatgaa attctcacca
     3541 tggaattaat gattatgaag gcccttaagt ggcgtttaag tcccctgact attgtgtcct
     3601 ggctgaatgt atacatgcag gttgcatatc taaatgactt acatgaagtg ctactgccgc
     3661 agtatcccca gcaaatcttt atacagattg cagagctgtt ggatctctgt gtcctggatg
     3721 ttgactgcct tgaatttcct tatggtatac ttgctgcttc ggccttgtat catttctcgt
     3781 catctgaatt gatgcaaaag gtttcagggt atcagtggtg cgacatagag aactgtgtca
     3841 agtggatggt tccatttgcc atggttataa gggagacggg gagctcaaaa ctgaagcact
     3901 tcaggggcgt cgctgatgaa gatgcacaca acatacagac ccacagagac agcttggatt
     3961 tgctggacaa agcccgagca aagaaagcca tgttgtctga acaaaatagg gcttctcctc
     4021 tccccagtgg gctcctcacc ccgccacaga gcggtaagaa gcagagcagc gggccggaaa
     4081 tggcgtgagg atcccgcgac tctagataat tctaccgggt aggggaggcg cttttcccaa
     4141 ggcagtctgg agcatgcgct ttagcagccc cgctgggcac ttggcgctac acaagtggcc
     4201 tctggcctcg cacacattcc acatccaccg gtaggcgcca accggctccg ttctttggtg
     4261 gccccttcgc gccaccttct actcctcccc tagtcaggaa gttccccccc gccccgcagc
     4321 tcgcgtcgtg caggacgtga caaatggaag tagcacgtct cactagtctc gtgcagatgg
     4381 acagcaccgc tgagcaatgg aagcgggtag gcctttgggg cagcggccaa tagcagcttt
     4441 gctccttcgc tttctgggct cagaggctgg gaaggggtgg gtccgggggc gggctcaggg
     4501 gcgggctcag gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc
     4561 ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc atctccgggc ctttcgacct
     4621 gcagcccaag cttaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     4681 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     4741 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     4801 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     4861 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     4921 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     4981 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     5041 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     5101 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     5161 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     5221 caagcccggt gcctgaccgc gtctggaaca atcaacctct ggattacaaa atttgtgaaa
     5281 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     5341 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     5401 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     5461 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     5521 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     5581 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     5641 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     5701 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     5761 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     5821 tttgggccgc ctccccgcct ggaattaatt ctgcagtcga gacctagaaa aacatggagc
     5881 aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag aagcacaaga
     5941 ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa tgacttacaa
     6001 ggcagctgta gatcttagcc actttttaaa agaaaagagg ggactggaag ggctaattca
     6061 ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag gctacttccc
     6121 tgattagcag aactacacac cagggccagg ggtcagatat ccactgacct ttggatggtg
     6181 ctacaagcta gtaccagttg agccagataa ggtagaagag gccaataaag gagagaacac
     6241 cagcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag aagtgttaga
     6301 gtggaggttt gacagccgcc tagcatttca tcacgtggcc cgagagctgc atccggagta
     6361 cttcaagaac tgctgatatc gagcttgcta caagggactt tccgctgggg actttccagg
     6421 gaggcgtggc ctgggcggga ctggggagtg gcgagccctc agatcctgca tataagcagc
     6481 tgctttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     6541 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     6601 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     6661 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
     6721 caaagaaatg aatatcagag agtgagaggc cttgacattg ctagcgtttt accgtcgacc
     6781 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     6841 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     6901 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     6961 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     7021 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     7081 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     7141 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     7201 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     7261 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     7321 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     7381 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     7441 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     7501 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     7561 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     7621 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     7681 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     7741 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     7801 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     7861 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     7921 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     7981 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     8041 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     8101 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     8161 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     8221 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     8281 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     8341 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     8401 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     8461 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     8521 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     8581 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     8641 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     8701 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     8761 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     8821 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     8881 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     8941 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc aacttgttta ttgcagctta
     9001 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     9061 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcaactg
     9121 gataactcaa gctaaccaaa atcatcccaa acttcccacc ccatacccta ttaccactgc
     9181 caattacctg tggtttcatt tactctaaac ctgtgattcc tctgaattat tttcatttta
     9241 aagaaattgt atttgttaaa tatgtactac aaacttagta gtttttaaag aaattgtatt
     9301 tgttaaatat gtactacaaa cttagtagt
//
