LOCUS       V019470                10003 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019470
VERSION     V019470
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             2843..4060
                     /label="CCBE1(NM_133459)"
                     /note="CCBE1(NM_133459)"
                     /gene="CCBE1"
     CDS             4091..4810
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="Aequorea coerulescens GFP"
                     /transl_table=1
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK*"
                     /label="AcGFP1"
     promoter        4838..5337
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5358..5957
                     /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    5971..6559
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6766..7399
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7528..7544)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7552..7568
                     /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(7576..7606)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7621..7642
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7930..8518)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8689..9549)
                     /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(9550..9654)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9702..9836
                     /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 gaattcgcca ccatggtgcc gccgcctccg agccggggag gagctgccag
     2881 gggccagctg ggcaggagcc tgggtccgct gctgctgctc ctggcgttgg gacacacgtg
     2941 gacctacaga gaggagccgg aggacggcga cagagaaatc tgctcagaga gcaaaatcgc
     3001 gacgactaaa tacccgtgtc tgaagtcttc aggcgagctc accacatgct acaggaaaaa
     3061 gtgctgcaaa ggatataaat ttgttcttgg acaatgcatc ccagaagatt acgacgtttg
     3121 tgccgaggct ccctgtgaac agcagtgcac ggacaacttt ggccgagtgc tgtgtacttg
     3181 ttatccggga taccgatatg accgggagag acaccggaag cgggagaagc catactgtct
     3241 ggatattgat gagtgtgcca gcagcaatgg gacgctgtgt gcccacatct gcatcaatac
     3301 cttgggcagc taccgctgcg agtgccggga aggctacatc cgggaagatg atgggaagac
     3361 atgtaccagg ggagacaaat atcccaatga cactggccat gagaagtctg agaacatggt
     3421 gaaagccgga acttgctgtg ccacatgcaa ggagttctac cagatgaagc agaccgtgct
     3481 gcagctgaag caaaagattg ctctgctccc caacaatgca gctgacctgg gcaagtatat
     3541 cactggtgac aaggtgctgg cctcaaacac ctaccttcca ggacctcctg gcctgcctgg
     3601 gggccagggc cctcccggct caccaggacc aaagggaagc ccaggcttcc ccggtatgcc
     3661 aggccctcct gggcagcccg gcccacgggg ctcaatggga cccatgggac catctcctga
     3721 tctgtcccac attaagcaag gccggagggg ccctgtgggt ccaccagggg caccaggaag
     3781 agatggttct aagggggaga gaggagcgcc tgggcccaga gggtctccag gaccccctgg
     3841 ttctttcgac ttcctgctac ttatgctggc tgacatccgc aatgacatca ctgagctgca
     3901 ggaaaaggtg ttcgggcacc ggactcactc ttcagcagag gagttccctt tacctcagga
     3961 atttcccagc tacccagaag ccatggacct gggctctgga gatgaccatc caagaagaac
     4021 tgagacaaga gacttgagag cccccagaga cttctaccca ggcggcggag ggagtatgga
     4081 tccaccggtc atggtgagca agggcgccga gctgttcacc ggcatcgtgc ccatcctgat
     4141 cgagctgaat ggcgatgtga atggccacaa gttcagcgtg agcggcgagg gcgagggcga
     4201 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcctgtgcc
     4261 ctggcccacc ctggtgacca ccctgagcta cggcgtgcag tgcttctcac gctaccccga
     4321 tcacatgaag cagcacgact tcttcaagag cgccatgcct gagggctaca tccaggagcg
     4381 caccatcttc ttcgaggatg acggcaacta caagtcgcgc gccgaggtga agttcgaggg
     4441 cgataccctg gtgaatcgca tcgagctgac cggcaccgat ttcaaggagg atggcaacat
     4501 cctgggcaat aagatggagt acaactacaa cgcccacaat gtgtacatca tgaccgacaa
     4561 ggccaagaat ggcatcaagg tgaacttcaa gatccgccac aacatcgagg atggcagcgt
     4621 gcagctggcc gaccactacc agcagaatac ccccatcggc gatggccctg tgctgctgcc
     4681 cgataaccac tacctgtcca cccagagcgc cctgtccaag gaccccaacg agaagcgcga
     4741 tcacatgatc tacttcggct tcgtgaccgc cgccgccatc acccacggca tggatgagct
     4801 gtacaagtga gcggccgcga ctctagataa ttctaccggg taggggaggc gcttttccca
     4861 aggcagtctg gagcatgcgc tttagcagcc ccgctgggca cttggcgcta cacaagtggc
     4921 ctctggcctc gcacacattc cacatccacc ggtaggcgcc aaccggctcc gttctttggt
     4981 ggccccttcg cgccaccttc tactcctccc ctagtcagga agttcccccc cgccccgcag
     5041 ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc tcactagtct cgtgcagatg
     5101 gacagcaccg ctgagcaatg gaagcgggta ggcctttggg gcagcggcca atagcagctt
     5161 tgctccttcg ctttctgggc tcagaggctg ggaaggggtg ggtccggggg cgggctcagg
     5221 ggcgggctca ggggcggggc gggcgcccga aggtcctccg gaggcccggc attctgcacg
     5281 cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct catctccggg cctttcgacc
     5341 tgcagcccaa gcttaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg
     5401 acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc
     5461 acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca
     5521 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg
     5581 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca
     5641 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc
     5701 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc
     5761 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg
     5821 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg
     5881 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc
     5941 gcaagcccgg tgcctgaccg cgtctggaac aatcaacctc tggattacaa aatttgtgaa
     6001 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     6061 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     6121 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     6181 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     6241 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     6301 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     6361 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
     6421 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     6481 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     6541 ctttgggccg cctccccgcc tggaattaat tctgcagtcg agacctagaa aaacatggag
     6601 caatcacaag tagcaataca gcagctacca atgctgattg tgcctggcta gaagcacaag
     6661 aggaggagga ggtgggtttt ccagtcacac ctcaggtacc tttaagacca atgacttaca
     6721 aggcagctgt agatcttagc cactttttaa aagaaaagag gggactggaa gggctaattc
     6781 actcccaacg aagacaagat atccttgatc tgtggatcta ccacacacaa ggctacttcc
     6841 ctgattagca gaactacaca ccagggccag gggtcagata tccactgacc tttggatggt
     6901 gctacaagct agtaccagtt gagccagata aggtagaaga ggccaataaa ggagagaaca
     6961 ccagcttgtt acaccctgtg agcctgcatg ggatggatga cccggagaga gaagtgttag
     7021 agtggaggtt tgacagccgc ctagcatttc atcacgtggc ccgagagctg catccggagt
     7081 acttcaagaa ctgctgatat cgagcttgct acaagggact ttccgctggg gactttccag
     7141 ggaggcgtgg cctgggcggg actggggagt ggcgagccct cagatcctgc atataagcag
     7201 ctgctttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     7261 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     7321 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     7381 gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt
     7441 gcaaagaaat gaatatcaga gagtgagagg ccttgacatt gctagcgttt taccgtcgac
     7501 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     7561 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     7621 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     7681 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     7741 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     7801 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     7861 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     7921 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     7981 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     8041 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     8101 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     8161 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     8221 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     8281 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     8341 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     8401 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     8461 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     8521 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     8581 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     8641 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     8701 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     8761 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     8821 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     8881 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     8941 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     9001 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     9061 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     9121 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     9181 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     9241 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     9301 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     9361 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     9421 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     9481 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     9541 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     9601 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     9661 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat caacttgttt attgcagctt
     9721 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     9781 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatcaact
     9841 ggataactca agctaaccaa aatcatccca aacttcccac cccataccct attaccactg
     9901 ccaattacct gtggtttcat ttactctaaa cctgtgattc ctctgaatta ttttcatttt
     9961 aaagaaattg tatttgttaa atatgtacta caaacttagt agt
//
