LOCUS       V040291                 9217 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040291
VERSION     V040291
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..3913
                     /label="GPRC5A(NM_003979)"
                     /note="GPRC5A(NM_003979)"
                     /gene="GPRC5A"
     CDS             3920..3985
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4005..4504
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4525..5124
                     /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    5138..5726
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5933..6566
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6695..6711)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6719..6735
                     /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(6743..6773)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6788..6809
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7097..7685)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7856..8716)
                     /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(8717..8821)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    8869..9003
                     /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 ccatggctac aacagtccct gatggttgcc gcaatggcct
     2881 gaaatccaag tactacagac tttgtgataa ggctgaagct tggggcatcg tcctagaaac
     2941 ggtggccaca gccggggttg tgacctcggt ggccttcatg ctcactctcc cgatcctcgt
     3001 ctgcaaggtg caggactcca acaggcgaaa aatgctgcct actcagtttc tcttcctcct
     3061 gggtgtgttg ggcatctttg gcctcacctt cgccttcatc atcggactgg acgggagcac
     3121 agggcccaca cgcttcttcc tctttgggat cctcttttcc atctgcttct cctgcctgct
     3181 ggctcatgct gtcagtctga ccaagctcgt ccgggggagg aagccccttt ccctgttggt
     3241 gattctgggt ctggccgtgg gcttcagcct agtccaggat gttatcgcta ttgaatatat
     3301 tgtcctgacc atgaatagga ccaacgtcaa tgtcttttct gagctttccg ctcctcgtcg
     3361 caatgaagac tttgtcctcc tgctcaccta cgtcctcttc ttgatggcgc tgaccttcct
     3421 catgtcctcc ttcaccttct gtggttcctt cacgggctgg aagagacatg gggcccacat
     3481 ctacctcacg atgctcctct ccattgccat ctgggtggcc tggatcaccc tgctcatgct
     3541 tcctgacttt gaccgcaggt gggatgacac catcctcagc tccgccttgg ctgccaatgg
     3601 ctgggtgttc ctgttggctt atgttagtcc cgagttttgg ctgctcacaa agcaacgaaa
     3661 ccccatggat tatcctgttg aggatgcttt ctgtaaacct caactcgtga agaagagcta
     3721 tggtgtggag aacagagcct actctcaaga ggaaatcact caaggttttg aagagacagg
     3781 ggacacgctc tatgccccct attccacaca ttttcagctg cagaaccagc ctccccaaaa
     3841 ggaattctcc atcccacggg cccacgcttg gccgagccct tacaaagact atgaagtaaa
     3901 gaaagagggc agcggatccg actacaaaga ccatgacggt gattataaag atcatgacat
     3961 cgactacaag gatgacgatg acaagtagtc tagataattc taccgggtag gggaggcgct
     4021 tttcccaagg cagtctggag catgcgcttt agcagccccg ctgggcactt ggcgctacac
     4081 aagtggcctc tggcctcgca cacattccac atccaccggt aggcgccaac cggctccgtt
     4141 ctttggtggc cccttcgcgc caccttctac tcctccccta gtcaggaagt tcccccccgc
     4201 cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca ctagtctcgt
     4261 gcagatggac agcaccgctg agcaatggaa gcgggtaggc ctttggggca gcggccaata
     4321 gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt ccgggggcgg
     4381 gctcaggggc gggctcaggg gcggggcggg cgcccgaagg tcctccggag gcccggcatt
     4441 ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc tcttcctcat ctccgggcct
     4501 ttcgacctgc agcccaagct taccatgacc gagtacaagc ccacggtgcg cctcgccacc
     4561 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
     4621 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
     4681 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
     4741 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
     4801 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc
     4861 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc
     4921 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag
     4981 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag
     5041 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
     5101 atgacccgca agcccggtgc ctgaccgcgt ctggaacaat caacctctgg attacaaaat
     5161 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     5221 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     5281 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     5341 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     5401 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     5461 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     5521 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
     5581 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     5641 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     5701 gatctccctt tgggccgcct ccccgcctgg aattaattct gcagtcgaga cctagaaaaa
     5761 catggagcaa tcacaagtag caatacagca gctaccaatg ctgattgtgc ctggctagaa
     5821 gcacaagagg aggaggaggt gggttttcca gtcacacctc aggtaccttt aagaccaatg
     5881 acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagagggg actggaaggg
     5941 ctaattcact cccaacgaag acaagatatc cttgatctgt ggatctacca cacacaaggc
     6001 tacttccctg attagcagaa ctacacacca gggccagggg tcagatatcc actgaccttt
     6061 ggatggtgct acaagctagt accagttgag ccagataagg tagaagaggc caataaagga
     6121 gagaacacca gcttgttaca ccctgtgagc ctgcatggga tggatgaccc ggagagagaa
     6181 gtgttagagt ggaggtttga cagccgccta gcatttcatc acgtggcccg agagctgcat
     6241 ccggagtact tcaagaactg ctgatatcga gcttgctaca agggactttc cgctggggac
     6301 tttccaggga ggcgtggcct gggcgggact ggggagtggc gagccctcag atcctgcata
     6361 taagcagctg ctttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga
     6421 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     6481 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     6541 ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt
     6601 ataacttgca aagaaatgaa tatcagagag tgagaggcct tgacattgct agcgttttac
     6661 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     6721 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     6781 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     6841 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     6901 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     6961 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     7021 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     7081 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     7141 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     7201 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     7261 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     7321 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     7381 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     7441 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     7501 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     7561 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     7621 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     7681 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     7741 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     7801 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     7861 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     7921 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     7981 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     8041 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     8101 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     8161 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     8221 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     8281 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     8341 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     8401 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     8461 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     8521 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     8581 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     8641 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     8701 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     8761 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     8821 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatcaa cttgtttatt
     8881 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     8941 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     9001 atcaactgga taactcaagc taaccaaaat catcccaaac ttcccacccc ataccctatt
     9061 accactgcca attacctgtg gtttcattta ctctaaacct gtgattcctc tgaattattt
     9121 tcattttaaa gaaattgtat ttgttaaata tgtactacaa acttagtagt ttttaaagaa
     9181 attgtatttg ttaaatatgt actacaaact tagtagt
//
