LOCUS       V034077                 9300 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034077
VERSION     V034077
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             2827..4056
                     /label="CCNE1(NM_001238)"
                     /note="CCNE1(NM_001238)"
                     /gene="CCNE1"
     promoter        4088..4587
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4608..5207
                     /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    5221..5809
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6016..6649
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6778..6794)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6802..6818
                     /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(6826..6856)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6871..6892
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7180..7768)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7939..8799)
                     /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(8800..8904)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    8952..9086
                     /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 gccaccatgc cgagggagcg cagggagcgg gatgcgaagg agcgggacac catgaaggag
     2881 gacggcggcg cggagttctc ggctcgctcc aggaagagga aggcaaacgt gaccgttttt
     2941 ttgcaggatc cagatgaaga aatggccaaa atcgacagga cggcgaggga ccagtgtggg
     3001 agccagcctt gggacaataa tgcagtctgt gcagacccct gctccctgat ccccacacct
     3061 gacaaagaag atgatgaccg ggtttaccca aactcaacgt gcaagcctcg gattattgca
     3121 ccatccagag gctccccgct gcctgtactg agctgggcaa atagagagga agtctggaaa
     3181 atcatgttaa acaaggaaaa gacatactta agggatcagc actttcttga gcaacaccct
     3241 cttctgcagc caaaaatgcg agcaattctt ctggattggt taatggaggt gtgtgaagtc
     3301 tataaacttc acagggagac cttttacttg gcacaagatt tctttgaccg gtatatggcg
     3361 acacaagaaa atgttgtaaa aactctttta cagcttattg ggatttcatc tttatttatt
     3421 gcagccaaac ttgaggaaat ctatcctcca aagttgcacc agtttgcgta tgtgacagat
     3481 ggagcttgtt caggagatga aattctcacc atggaattaa tgattatgaa ggcccttaag
     3541 tggcgtttaa gtcccctgac tattgtgtcc tggctgaatg tatacatgca ggttgcatat
     3601 ctaaatgact tacatgaagt gctactgccg cagtatcccc agcaaatctt tatacagatt
     3661 gcagagctgt tggatctctg tgtcctggat gttgactgcc ttgaatttcc ttatggtata
     3721 cttgctgctt cggccttgta tcatttctcg tcatctgaat tgatgcaaaa ggtttcaggg
     3781 tatcagtggt gcgacataga gaactgtgtc aagtggatgg ttccatttgc catggttata
     3841 agggagacgg ggagctcaaa actgaagcac ttcaggggcg tcgctgatga agatgcacac
     3901 aacatacaga cccacagaga cagcttggat ttgctggaca aagcccgagc aaagaaagcc
     3961 atgttgtctg aacaaaatag ggcttctcct ctccccagtg ggctcctcac cccgccacag
     4021 agcggtaaga agcagagcag cgggccggaa atggcgtgag gatcccgcga ctctagataa
     4081 ttctaccggg taggggaggc gcttttccca aggcagtctg gagcatgcgc tttagcagcc
     4141 ccgctgggca cttggcgcta cacaagtggc ctctggcctc gcacacattc cacatccacc
     4201 ggtaggcgcc aaccggctcc gttctttggt ggccccttcg cgccaccttc tactcctccc
     4261 ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa
     4321 gtagcacgtc tcactagtct cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta
     4381 ggcctttggg gcagcggcca atagcagctt tgctccttcg ctttctgggc tcagaggctg
     4441 ggaaggggtg ggtccggggg cgggctcagg ggcgggctca ggggcggggc gggcgcccga
     4501 aggtcctccg gaggcccggc attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc
     4561 tcctcttcct catctccggg cctttcgacc tgcagcccaa gcttaccatg accgagtaca
     4621 agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg
     4681 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc
     4741 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
     4801 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
     4861 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
     4921 cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
     4981 tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
     5041 tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
     5101 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
     5161 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgaccg cgtctggaac
     5221 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     5281 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     5341 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     5401 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     5461 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     5521 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     5581 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc
     5641 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     5701 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     5761 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaattaat
     5821 tctgcagtcg agacctagaa aaacatggag caatcacaag tagcaataca gcagctacca
     5881 atgctgattg tgcctggcta gaagcacaag aggaggagga ggtgggtttt ccagtcacac
     5941 ctcaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
     6001 aagaaaagag gggactggaa gggctaattc actcccaacg aagacaagat atccttgatc
     6061 tgtggatcta ccacacacaa ggctacttcc ctgattagca gaactacaca ccagggccag
     6121 gggtcagata tccactgacc tttggatggt gctacaagct agtaccagtt gagccagata
     6181 aggtagaaga ggccaataaa ggagagaaca ccagcttgtt acaccctgtg agcctgcatg
     6241 ggatggatga cccggagaga gaagtgttag agtggaggtt tgacagccgc ctagcatttc
     6301 atcacgtggc ccgagagctg catccggagt acttcaagaa ctgctgatat cgagcttgct
     6361 acaagggact ttccgctggg gactttccag ggaggcgtgg cctgggcggg actggggagt
     6421 ggcgagccct cagatcctgc atataagcag ctgctttttg cctgtactgg gtctctctgg
     6481 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     6541 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     6601 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca
     6661 tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg
     6721 ccttgacatt gctagcgttt taccgtcgac ctctagctag agcttggcgt aatcatggtc
     6781 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     6841 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     6901 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     6961 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     7021 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     7081 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     7141 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     7201 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     7261 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     7321 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     7381 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     7441 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     7501 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     7561 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     7621 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     7681 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     7741 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     7801 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     7861 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     7921 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7981 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     8041 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     8101 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     8161 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     8221 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     8281 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     8341 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     8401 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     8461 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     8521 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     8581 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     8641 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     8701 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     8761 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     8821 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     8881 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     8941 atcgggagat caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     9001 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     9061 tcaatgtatc ttatcatgtc tggatcaact ggataactca agctaaccaa aatcatccca
     9121 aacttcccac cccataccct attaccactg ccaattacct gtggtttcat ttactctaaa
     9181 cctgtgattc ctctgaatta ttttcatttt aaagaaattg tatttgttaa atatgtacta
     9241 caaacttagt agtttttaaa gaaattgtat ttgttaaata tgtactacaa acttagtagt
//
