LOCUS       V018283                10086 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018283
VERSION     V018283
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             2807..3523
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="Aequorea coerulescens GFP"
                     /transl_table=1
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
                     /label="AcGFP1"
     CDS             3545..4879
                     /label="NEK2(NM_002497)"
                     /note="NEK2(NM_002497)"
                     /gene="NEK2"
     promoter        4921..5420
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5441..6040
                     /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    6054..6642
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6849..7482
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7611..7627)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7635..7651
                     /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(7659..7689)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7704..7725
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8013..8601)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8772..9632)
                     /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(9633..9737)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9785..9919
                     /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 tctaccggtc gccaccatgg tgagcaaggg
     2821 cgccgagctg ttcaccggca tcgtgcccat cctgatcgag ctgaatggcg atgtgaatgg
     2881 ccacaagttc agcgtgagcg gcgagggcga gggcgatgcc acctacggca agctgaccct
     2941 gaagttcatc tgcaccaccg gcaagctgcc tgtgccctgg cccaccctgg tgaccaccct
     3001 gagctacggc gtgcagtgct tctcacgcta ccccgatcac atgaagcagc acgacttctt
     3061 caagagcgcc atgcctgagg gctacatcca ggagcgcacc atcttcttcg aggatgacgg
     3121 caactacaag tcgcgcgccg aggtgaagtt cgagggcgat accctggtga atcgcatcga
     3181 gctgaccggc accgatttca aggaggatgg caacatcctg ggcaataaga tggagtacaa
     3241 ctacaacgcc cacaatgtgt acatcatgac cgacaaggcc aagaatggca tcaaggtgaa
     3301 cttcaagatc cgccacaaca tcgaggatgg cagcgtgcag ctggccgacc actaccagca
     3361 gaataccccc atcggcgatg gccctgtgct gctgcccgat aaccactacc tgtccaccca
     3421 gagcgccctg tccaaggacc ccaacgagaa gcgcgatcac atgatctact tcggcttcgt
     3481 gaccgccgcc gccatcaccc acggcatgga tgagctgtac aagtccggac tcagatctcg
     3541 agtcatgcct tcccgggctg aggactatga agtgttgtac accattggca caggctccta
     3601 cggccgctgc cagaagatcc ggaggaagag tgatggcaag atattagttt ggaaagaact
     3661 tgactatggc tccatgacag aagctgagaa acagatgctt gtttctgaag tgaatttgct
     3721 tcgtgaactg aaacatccaa acatcgttcg ttactatgat cggattattg accggaccaa
     3781 tacaacactg tacattgtaa tggaatattg tgaaggaggg gatctggcta gtgtaattac
     3841 aaagggaacc aaggaaaggc aatacttaga tgaagagttt gttcttcgag tgatgactca
     3901 gttgactctg gccctgaagg aatgccacag acgaagtgat ggtggtcata ccgtattgca
     3961 tcgggatctg aaaccagcca atgttttcct ggatggcaag caaaacgtca agcttggaga
     4021 ctttgggcta gctagaatat taaaccatga cacgagtttt gcaaaaacat ttgttggcac
     4081 accttattac atgtctcctg aacaaatgaa tcgcatgtcc tacaatgaga aatcagatat
     4141 ctggtcattg ggctgcttgc tgtatgagtt atgtgcatta atgcctccat ttacagcttt
     4201 tagccagaaa gaactcgctg ggaaaatcag agaaggcaaa ttcaggcgaa ttccataccg
     4261 ttactctgat gaattgaatg aaattattac gaggatgtta aacttaaagg attaccatcg
     4321 accttctgtt gaagaaattc ttgagaaccc tttaatagca gatttggttg cagacgagca
     4381 aagaagaaat cttgagagaa gagggcgaca attaggagag ccagaaaaat cgcaggattc
     4441 cagccctgta ttgagtgagc tgaaactgaa ggaaattcag ttacaggagc gagagcgagc
     4501 tctcaaagca agagaagaaa gattggagca gaaagaacag gagctttgtg ttcgtgagag
     4561 actagcagag gacaaactgg ctagagcaga aaatctgttg aagaactaca gcttgctaaa
     4621 ggaacggaag ttcctgtctc tggcaagtaa tccagaactt cttaatcttc catcctcagt
     4681 aattaagaag aaagttcatt tcagtgggga aagtaaagag aacatcatga ggagtgagaa
     4741 ttctgagagt cagctcacat ctaagtccaa gtgcaaggac ctgaagaaaa ggcttcacgc
     4801 tgcccagctg cgggctcaag ccctgtcaga tattgagaaa aattaccaac tgaaaagcag
     4861 acagatcctg ggcatgcgct agggatccac cggatctaga taactgatca taattctacc
     4921 gggtagggga ggcgcttttc ccaaggcagt ctggagcatg cgctttagca gccccgctgg
     4981 gcacttggcg ctacacaagt ggcctctggc ctcgcacaca ttccacatcc accggtaggc
     5041 gccaaccggc tccgttcttt ggtggcccct tcgcgccacc ttctactcct cccctagtca
     5101 ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac
     5161 gtctcactag tctcgtgcag atggacagca ccgctgagca atggaagcgg gtaggccttt
     5221 ggggcagcgg ccaatagcag ctttgctcct tcgctttctg ggctcagagg ctgggaaggg
     5281 gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct
     5341 ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt
     5401 cctcatctcc gggcctttcg acctgcagcc caagcttacc atgaccgagt acaagcccac
     5461 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt
     5521 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac
     5581 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     5641 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     5701 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     5761 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     5821 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     5881 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     5941 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     6001 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga ccgcgtctgg aacaatcaac
     6061 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     6121 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     6181 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     6241 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     6301 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     6361 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     6421 ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg
     6481 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     6541 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     6601 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaatt aattctgcag
     6661 tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga
     6721 ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt
     6781 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     6841 gaggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat
     6901 ctaccacaca caaggctact tccctgatta gcagaactac acaccagggc caggggtcag
     6961 atatccactg acctttggat ggtgctacaa gctagtacca gttgagccag ataaggtaga
     7021 agaggccaat aaaggagaga acaccagctt gttacaccct gtgagcctgc atgggatgga
     7081 tgacccggag agagaagtgt tagagtggag gtttgacagc cgcctagcat ttcatcacgt
     7141 ggcccgagag ctgcatccgg agtacttcaa gaactgctga tatcgagctt gctacaaggg
     7201 actttccgct ggggactttc cagggaggcg tggcctgggc gggactgggg agtggcgagc
     7261 cctcagatcc tgcatataag cagctgcttt ttgcctgtac tgggtctctc tggttagacc
     7321 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     7381 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     7441 gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc
     7501 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggccttgac
     7561 attgctagcg ttttaccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     7621 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     7681 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     7741 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     7801 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     7861 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     7921 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     7981 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     8041 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     8101 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     8161 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     8221 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     8281 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     8341 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     8401 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     8461 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     8521 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     8581 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     8641 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     8701 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     8761 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     8821 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     8881 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     8941 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     9001 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     9061 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     9121 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     9181 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     9241 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     9301 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     9361 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     9421 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     9481 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     9541 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     9601 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     9661 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     9721 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     9781 gatcaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     9841 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     9901 atcttatcat gtctggatca actggataac tcaagctaac caaaatcatc ccaaacttcc
     9961 caccccatac cctattacca ctgccaatta cctgtggttt catttactct aaacctgtga
    10021 ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt taaatatgta ctacaaactt
    10081 agtagt
//
