LOCUS       V041024                10120 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041024
VERSION     V041024
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             2840..2866
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2873..4135
                     /label="Acadm(NM_007382)"
                     /note="Acadm(NM_007382)"
                     /gene="Acadm"
     CDS             4172..4891
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK*"
                     /label="EGFP"
     promoter        4908..5407
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5428..6027
                     /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    6041..6629
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6836..7469
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7598..7614)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7622..7638
                     /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(7646..7676)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7691..7712
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8000..8588)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8759..9619)
                     /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(9620..9724)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9772..9906
                     /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 ctcaagcttc gccaccatgt acccatacga tgttccagat tacgctgaat tcatggcagc
     2881 ggcgttccgc agaggctgca gggtcctgag aagtgtttct cattttgagt gtcgaacaca
     2941 acactcgaaa gcggctcaca agcaggagcc cggattaggg tttagttttg agttgacgga
     3001 acagcagaaa gagtttcaag caactgcccg caagtttgcc agagaggaga ttatccccgt
     3061 cgccccggaa tatgacaaaa gcggggagta cccgttccct ctcatcaaaa gagcctggga
     3121 actcggcttg atcaacgcgc acattccgga aagttgcggt ggccttggcc tgggaacgtt
     3181 cgatgcttgt ttaattaccg aagagttggc gtatgggtgt acaggggtgc aaactgctat
     3241 tgaagcaaat tctttggggc aaatgcctgt gattcttgct ggaaatgatc aacaaaaaaa
     3301 gaagtatttg gggaggatga cggagcagcc aatgatgtgt gcttactgtg tgacagagcc
     3361 ctccgcaggc tctgatgtgg cggccattaa gaccaaagca gagaagaagg gtgacgagta
     3421 tgttatcaac ggccagaaga tgtggataac caacggggga aaggccaact ggtatttctt
     3481 gttggcacgt tctaacccag atcctaaagt acccgctagt aaagccttta ctggattcat
     3541 tgtggaagcc gacaccccgg gaatacatat tggaaaaaag gaactaaaca tgggccagcg
     3601 atgctctgac accagaggaa ttgctttcga agacgtcaga gtgcctaagg aaaatgtgtt
     3661 aatcggtgaa ggagcaggtt tcaagatcgc aatgggtgct tttgatagaa ccagacctac
     3721 agtcgcagct ggcgctgtcg ggctagccca gagagctcta gacgaagcca cgaagtatgc
     3781 cctggatagg aagacatttg gaaagctgct agtggagcac caaggagttt catttctgct
     3841 cgcagaaatg gcgatgaagg ttgaactcgc taggctcagt taccagagag cagcctggga
     3901 ggttgactcc ggtcgccgga acacttacta tgcctcgatt gcaaaggcct ttgctggaga
     3961 cattgccaat cagctagcca ctgacgccgt gcagattttc ggaggctatg gattcaacac
     4021 agagtaccct gtggagaagc tgatgaggga cgccaagatc tatcagattt atgaaggtac
     4081 tgcacaaatt cagaggctga tcatagctcg tgagcacatt gaaaagtata aaaatggtgg
     4141 aggcggatca ggaggcggtg gatccgccac catggtgagc aagggcgagg agctgttcac
     4201 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     4261 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     4321 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     4381 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     4441 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     4501 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     4561 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     4621 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     4681 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     4741 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     4801 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     4861 cactctcggc atggacgagc tgtacaagta gtctagataa ttctaccggg taggggaggc
     4921 gcttttccca aggcagtctg gagcatgcgc tttagcagcc ccgctgggca cttggcgcta
     4981 cacaagtggc ctctggcctc gcacacattc cacatccacc ggtaggcgcc aaccggctcc
     5041 gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga agttcccccc
     5101 cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc tcactagtct
     5161 cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg gcagcggcca
     5221 atagcagctt tgctccttcg ctttctgggc tcagaggctg ggaaggggtg ggtccggggg
     5281 cgggctcagg ggcgggctca ggggcggggc gggcgcccga aggtcctccg gaggcccggc
     5341 attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct catctccggg
     5401 cctttcgacc tgcagcccaa gcttaccatg accgagtaca agcccacggt gcgcctcgcc
     5461 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
     5521 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa
     5581 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
     5641 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
     5701 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
     5761 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg
     5821 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc
     5881 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
     5941 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
     6001 tgcatgaccc gcaagcccgg tgcctgaccg cgtctggaac aatcaacctc tggattacaa
     6061 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     6121 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     6181 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     6241 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     6301 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     6361 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     6421 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
     6481 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     6541 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     6601 tcggatctcc ctttgggccg cctccccgcc tggaattaat tctgcagtcg agacctagaa
     6661 aaacatggag caatcacaag tagcaataca gcagctacca atgctgattg tgcctggcta
     6721 gaagcacaag aggaggagga ggtgggtttt ccagtcacac ctcaggtacc tttaagacca
     6781 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaagag gggactggaa
     6841 gggctaattc actcccaacg aagacaagat atccttgatc tgtggatcta ccacacacaa
     6901 ggctacttcc ctgattagca gaactacaca ccagggccag gggtcagata tccactgacc
     6961 tttggatggt gctacaagct agtaccagtt gagccagata aggtagaaga ggccaataaa
     7021 ggagagaaca ccagcttgtt acaccctgtg agcctgcatg ggatggatga cccggagaga
     7081 gaagtgttag agtggaggtt tgacagccgc ctagcatttc atcacgtggc ccgagagctg
     7141 catccggagt acttcaagaa ctgctgatat cgagcttgct acaagggact ttccgctggg
     7201 gactttccag ggaggcgtgg cctgggcggg actggggagt ggcgagccct cagatcctgc
     7261 atataagcag ctgctttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg
     7321 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     7381 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     7441 cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta
     7501 tttataactt gcaaagaaat gaatatcaga gagtgagagg ccttgacatt gctagcgttt
     7561 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     7621 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     7681 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     7741 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     7801 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     7861 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     7921 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     7981 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     8041 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     8101 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     8161 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     8221 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     8281 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     8341 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     8401 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     8461 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     8521 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     8581 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     8641 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     8701 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     8761 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     8821 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     8881 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     8941 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     9001 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     9061 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     9121 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     9181 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     9241 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     9301 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     9361 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     9421 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     9481 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     9541 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     9601 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     9661 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     9721 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat caacttgttt
     9781 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     9841 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     9901 tggatcaact ggataactca agctaaccaa aatcatccca aacttcccac cccataccct
     9961 attaccactg ccaattacct gtggtttcat ttactctaaa cctgtgattc ctctgaatta
    10021 ttttcatttt aaagaaattg tatttgttaa atatgtacta caaacttagt agtttttaaa
    10081 gaaattgtat ttgttaaata tgtactacaa acttagtagt
//
