LOCUS       V035151                 9865 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035151
VERSION     V035151
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..4561
                     /label="Acsm1(NM_054094)"
                     /note="Acsm1(NM_054094)"
                     /gene="Acsm1"
     CDS             4568..4633
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4653..5152
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5173..5772
                     /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    5786..6374
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6581..7214
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7343..7359)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7367..7383
                     /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(7391..7421)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7436..7457
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7745..8333)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8504..9364)
                     /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(9365..9469)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9517..9651
                     /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 ccatgcagtg gctgaagagt ttccagatct gcaaagtctt
     2881 acagggcttc tccctaagcc ccacccagct gcacagacgg ttgttttcaa gagttggagc
     2941 tccaagatgg aatgaccatg attcacctga ggagtttaac tttgcaagtg atgtcctgga
     3001 ctactgggct caaatggagg aggagggcaa gagaggacca agtccagcct tttggtgggt
     3061 gaatggccaa ggagatgaaa taaagtggag cttcaggaag ctgagggacc tcacctgtcg
     3121 cactgccaac gtctttgagc agatttgtgg cctgcagcaa ggagatcacc tggccttgat
     3181 tctgccccga gtgcccgagt ggtggttggt gacagtgggc tgcatgcgaa cagggatcat
     3241 cttcatgcct gggactaccc aactgaaagc caaggacatt ctctaccgaa tacaaatatc
     3301 tcgagccaaa gccattgtga ccacagctag ccttgtccca gaggtggaat ctgtggcttc
     3361 tgagtgtcct gatctgaaaa ccaagctggt ggtgtctgat cacagccatg aagggtggct
     3421 tgatttctgt tcactgatta aatcagcatc cccagaccat acttgtatta agtcaaagat
     3481 gaaggatccc atggccatct tcttcaccag tgggaccaca ggctacccca agatggcaaa
     3541 gcacaaccag ggacttgcct tccggtcata tatcccttca tgcagaaaat tattgaagct
     3601 gaagacatct gacatcttgt ggtgcatgtc agacccagga tggattctgg ctaccgtggg
     3661 gtgcctgatc gagccatgga catcaggatg tacagtcttc atccaccacc tccctcaatt
     3721 cgaccccaaa gtcattgtag aggtactgtt caaatacccc atcactcagt gccttgctgc
     3781 cccaggcgtg tatcgaatgg ttcttcagca gaaaacctcc aacctcaggt tccccaccct
     3841 tgagcattgc actactggtg gggagagcct gctgcctgag gagtatgagc agtggaagca
     3901 aaggacaggt ctttccatcc acgaggtcta tggacagtca gaaacgggga tcagcagtgc
     3961 caccctccgg gaaatgaaga tcaagcgagg ctccataggg aaggccatct taccctttga
     4021 cttgcagatc atcgatgaaa agggcaacat cctcccaccc aacactgaag gatacattgg
     4081 catcaggatc aagcccacca ggcctctagg cctcttcatg gaatatgaga atagcccaga
     4141 gagcacatct gaagtggagt gtggggactt ttacaatagt ggggatagag cgaccattga
     4201 tgaagagggc tacatctggt tcttgggaag gggcgatgat gtcatcaatg cttccgggta
     4261 tcgcatcggg cctgtagagg tggagaacgc cttggcggag cacccagcag tggcagagtc
     4321 tgcggtggtg agcagcccgg acaaggatcg aggagaggtg gtgaaggcgt ttattgtcct
     4381 caacccagag ttcctgtcac acgatcagga acagcttatc aaagagctac agcatcatgt
     4441 gaagtcagtg acggcaccat acaagtaccc caggaaggtg gagtttgttt cagaattgcc
     4501 caaaactgtc acaggcaaaa tcaaaaggaa ggaacttcga aacaaggagt ttggtcagct
     4561 aggatccgac tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga
     4621 tgacgatgac aagtagtcta gataattcta ccgggtaggg gaggcgcttt tcccaaggca
     4681 gtctggagca tgcgctttag cagccccgct gggcacttgg cgctacacaa gtggcctctg
     4741 gcctcgcaca cattccacat ccaccggtag gcgccaaccg gctccgttct ttggtggccc
     4801 cttcgcgcca ccttctactc ctcccctagt caggaagttc ccccccgccc cgcagctcgc
     4861 gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact agtctcgtgc agatggacag
     4921 caccgctgag caatggaagc gggtaggcct ttggggcagc ggccaatagc agctttgctc
     4981 cttcgctttc tgggctcaga ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg
     5041 gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc ccggcattct gcacgcttca
     5101 aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct ccgggccttt cgacctgcag
     5161 cccaagctta ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     5221 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     5281 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     5341 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     5401 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     5461 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     5521 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     5581 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     5641 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     5701 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     5761 cccggtgcct gaccgcgtct ggaacaatca acctctggat tacaaaattt gtgaaagatt
     5821 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     5881 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     5941 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     6001 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     6061 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     6121 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     6181 gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     6241 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     6301 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     6361 ggccgcctcc ccgcctggaa ttaattctgc agtcgagacc tagaaaaaca tggagcaatc
     6421 acaagtagca atacagcagc taccaatgct gattgtgcct ggctagaagc acaagaggag
     6481 gaggaggtgg gttttccagt cacacctcag gtacctttaa gaccaatgac ttacaaggca
     6541 gctgtagatc ttagccactt tttaaaagaa aagaggggac tggaagggct aattcactcc
     6601 caacgaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta cttccctgat
     6661 tagcagaact acacaccagg gccaggggtc agatatccac tgacctttgg atggtgctac
     6721 aagctagtac cagttgagcc agataaggta gaagaggcca ataaaggaga gaacaccagc
     6781 ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt gttagagtgg
     6841 aggtttgaca gccgcctagc atttcatcac gtggcccgag agctgcatcc ggagtacttc
     6901 aagaactgct gatatcgagc ttgctacaag ggactttccg ctggggactt tccagggagg
     6961 cgtggcctgg gcgggactgg ggagtggcga gccctcagat cctgcatata agcagctgct
     7021 ttttgcctgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     7081 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     7141 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     7201 gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa
     7261 gaaatgaata tcagagagtg agaggccttg acattgctag cgttttaccg tcgacctcta
     7321 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     7381 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     7441 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     7501 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     7561 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     7621 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     7681 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     7741 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     7801 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     7861 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     7921 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     7981 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     8041 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     8101 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     8161 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     8221 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     8281 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     8341 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     8401 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     8461 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     8521 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     8581 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     8641 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     8701 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     8761 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     8821 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     8881 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     8941 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     9001 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     9061 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     9121 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     9181 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     9241 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     9301 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     9361 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     9421 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     9481 gaaaagtgcc acctgacgtc gacggatcgg gagatcaact tgtttattgc agcttataat
     9541 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     9601 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat caactggata
     9661 actcaagcta accaaaatca tcccaaactt cccaccccat accctattac cactgccaat
     9721 tacctgtggt ttcatttact ctaaacctgt gattcctctg aattattttc attttaaaga
     9781 aattgtattt gttaaatatg tactacaaac ttagtagttt ttaaagaaat tgtatttgtt
     9841 aaatatgtac tacaaactta gtagt
//
