LOCUS       pCMV-Puro-APP(human)    7502 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019308
VERSION     V019308
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             908..3217
                     /label="APP(NM_000484)"
                     /note="APP(NM_000484)"
                     /gene="APP"
     polyA_signal    3264..3488
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3534..3962
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3976..4305
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4372..4971
                     /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"
     polyA_signal    5145..5266
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5315..5331)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5339..5355
                     /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(5363..5393)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5408..5429
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5717..6302)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6473..7333)
                     /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(7334..7438)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cgccaccatg ctgcccggtt tggcactgct cctgctggcc gcctggacgg ctcgggcgct
      961 ggaggtaccc actgatggta atgctggcct gctggctgaa ccccagattg ccatgttctg
     1021 tggcagactg aacatgcaca tgaatgtcca gaatgggaag tgggattcag atccatcagg
     1081 gaccaaaacc tgcattgata ccaaggaagg catcctgcag tattgccaag aagtctaccc
     1141 tgaactgcag atcaccaatg tggtagaagc caaccaacca gtgaccatcc agaactggtg
     1201 caagcggggc cgcaagcagt gcaagaccca tccccacttt gtgattccct accgctgctt
     1261 agttggtgag tttgtaagtg atgcccttct cgttcctgac aagtgcaaat tcttacacca
     1321 ggagaggatg gatgtttgcg aaactcatct tcactggcac accgtcgcca aagagacatg
     1381 cagtgagaag agtaccaact tgcatgacta cggcatgttg ctgccctgcg gaattgacaa
     1441 gttccgaggg gtagagtttg tgtgttgccc actggctgaa gaaagtgaca atgtggattc
     1501 tgctgatgcg gaggaggatg actcggatgt ctggtggggc ggagcagaca cagactatgc
     1561 agatgggagt gaagacaaag tagtagaagt agcagaggag gaagaagtgg ctgaggtgga
     1621 agaagaagaa gccgatgatg acgaggacga tgaggatggt gatgaggtag aggaagaggc
     1681 tgaggaaccc tacgaagaag ccacagagag aaccaccagc attgccacca ccaccaccac
     1741 caccacagag tctgtggaag aggtggttcg agaggtgtgc tctgaacaag ccgagacggg
     1801 gccgtgccga gcaatgatct cccgctggta ctttgatgtg actgaaggga agtgtgcccc
     1861 attcttttac ggcggatgtg gcggcaaccg gaacaacttt gacacagaag agtactgcat
     1921 ggccgtgtgt ggcagcgcca tgtcccaaag tttactcaag actacccagg aacctcttgc
     1981 ccgagatcct gttaaacttc ctacaacagc agccagtacc cctgatgccg ttgacaagta
     2041 tctcgagaca cctggggatg agaatgaaca tgcccatttc cagaaagcca aagagaggct
     2101 tgaggccaag caccgagaga gaatgtccca ggtcatgaga gaatgggaag aggcagaacg
     2161 tcaagcaaag aacttgccta aagctgataa gaaggcagtt atccagcatt tccaggagaa
     2221 agtggaatct ttggaacagg aagcagccaa cgagagacag cagctggtgg agacacacat
     2281 ggccagagtg gaagccatgc tcaatgaccg ccgccgcctg gccctggaga actacatcac
     2341 cgctctgcag gctgttcctc ctcggcctcg tcacgtgttc aatatgctaa agaagtatgt
     2401 ccgcgcagaa cagaaggaca gacagcacac cctaaagcat ttcgagcatg tgcgcatggt
     2461 ggatcccaag aaagccgctc agatccggtc ccaggttatg acacacctcc gtgtgattta
     2521 tgagcgcatg aatcagtctc tctccctgct ctacaacgtg cctgcagtgg ccgaggagat
     2581 tcaggatgaa gttgatgagc tgcttcagaa agagcaaaac tattcagatg acgtcttggc
     2641 caacatgatt agtgaaccaa ggatcagtta cggaaacgat gctctcatgc catctttgac
     2701 cgaaacgaaa accaccgtgg agctccttcc cgtgaatgga gagttcagcc tggacgatct
     2761 ccagccgtgg cattcttttg gggctgactc tgtgccagcc aacacagaaa acgaagttga
     2821 gcctgttgat gcccgccctg ctgccgaccg aggactgacc actcgaccag gttctgggtt
     2881 gacaaatatc aagacggagg agatctctga agtgaagatg gatgcagaat tccgacatga
     2941 ctcaggatat gaagttcatc atcaaaaatt ggtgttcttt gcagaagatg tgggttcaaa
     3001 caaaggtgca atcattggac tcatggtggg cggtgttgtc atagcgacag tgatcgtcat
     3061 caccttggtg atgctgaaga agaaacagta cacatccatt catcatggtg tggtggaggt
     3121 tgacgccgct gtcaccccag aggagcgcca cctgtccaag atgcagcaga acggctacga
     3181 aaatccaacc tacaagttct ttgagcagat gcagaactag ctcgagtcta gagggcccgt
     3241 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     3301 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     3361 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     3421 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     3481 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     3541 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3601 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3661 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3721 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3781 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3841 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3901 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3961 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4021 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4081 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4141 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4201 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     4261 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     4321 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgaccgag
     4381 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt acgcaccctc
     4441 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgacccgga ccgccacatc
     4501 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     4561 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     4621 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     4681 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     4741 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     4801 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc
     4861 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     4921 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg agcgggactc
     4981 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     5041 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     5101 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     5161 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5221 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     5281 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     5341 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     5401 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     5461 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5521 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5581 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5641 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5701 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5761 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5821 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5881 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     5941 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6001 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6061 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6121 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     6181 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6241 ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6301 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6361 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6421 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6481 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6541 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6601 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6661 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6721 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6781 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6841 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6901 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6961 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7021 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7081 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7141 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7201 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     7261 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7321 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     7381 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7441 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     7501 gt
//
