LOCUS       V027130                 7760 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027130
VERSION     V027130
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             941..3250
                     /label="App(NM_001198823)"
                     /note="App(NM_001198823)"
                     /gene="App"
     CDS             3257..3322
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3360..3584
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3630..4058
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4072..4401
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4468..5262
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    5436..5557
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5606..5622)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5630..5646
                     /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(5654..5684)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5699..5720
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6008..6593)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6764..7624)
                     /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(7625..7729)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgctgccca gcttggcact
      961 gctcctgctg gccgcctgga cggttcgggc tctggaggta cccactgatg gcaacgccgg
     1021 gctgctggca gaaccccaga tcgccatgtt ctgtggtaaa ctcaacatgc acatgaatgt
     1081 gcagaatgga aagtgggagt cagacccgtc agggaccaaa acctgcattg gcaccaagga
     1141 gggcatcttg cagtactgcc aagaggtcta ccctgaactg cagatcacaa acgtggtgga
     1201 agccaaccag ccagtgacca tccagaactg gtgcaagcgg ggccgcaagc agtgcaagac
     1261 acacacccac atcgtgattc cttaccgttg cctagttggt gagtttgtga gcgacgccct
     1321 tctcgtgccc gacaagtgca agttcctaca ccaggagcgg atggatgttt gtgagaccca
     1381 tcttcactgg cacaccgtcg ccaaagagac atgcagcgag aagagcacta acttgcacga
     1441 ctatggcatg ctgctgccct gcggcatcga caagttccga ggggtagagt ttgtatgctg
     1501 cccgttggcc gaggaaagcg acagcgtgga ttctgcggat gcagaggagg atgactctga
     1561 tgtctggtgg ggtggagcgg acacagacta cgctgatggc ggtgaagaca aagtagtaga
     1621 agtcgccgaa gaggaggaag tggctgatgt tgaggaagag gaagctgatg atgatgagga
     1681 tgtggaggat ggggacgagg tggaggagga ggccgaggag ccctacgaag aggccaccga
     1741 gagaacaacc agcactgcca ccaccaccac aaccaccact gagtccgtgg aggaggtggt
     1801 ccgagaggtg tgctctgaac aagccgagac cgggccatgc cgcgcaatga tctcccgctg
     1861 gtactttgat gtcactgaag ggaagtgtgt cccattcttt tacggcggat gtggcggcaa
     1921 caggaacaac tttgacacgg aagagtactg catggcggtg tgtggcagcg tgtcaaccca
     1981 aagtttactc aagactacca gtgaacctct tccccaagat cctgataaac ttcccacgac
     2041 agcagccagc acccccgacg ccgtcgacaa gtacctggag acacccgggg acgagaacga
     2101 gcatgcccat ttccagaaag ccaaagagag gctggaagcc aagcaccgag agagaatgtc
     2161 ccaggtcatg agagaatggg aagaggcaga gcgtcaagcc aagaacttgc ccaaagctga
     2221 caagaaggcc gttatccagc atttccagga gaaagtggaa tctctggaac aggaagcagc
     2281 caatgagaga cagcagcttg tagagacaca catggccaga gttgaagcca tgctcaatga
     2341 ccgccgccgc ctggccctcg agaattacat cactgcactg caggcggtgc ccccaaggcc
     2401 tcatcatgtg ttcaacatgc tgaagaagta cgtccgtgcg gagcagaaag acagacagca
     2461 caccctaaag cattttgaac atgtgcgcat ggtggacccc aagaaagctg ctcagatccg
     2521 gtcccaggtt atgacacacc tccgtgtgat ctacgagcgc atgaaccagt ctctgtccct
     2581 gctctacaat gtccctgcgg tggctgagga gattcaagat gaagtcgatg agctgcttca
     2641 gaaggagcag aactactccg acgatgtctt ggccaacatg atcagtgagc ccagaatcag
     2701 ctacggaaac gacgctctca tgccttcgct gacggaaacc aagaccaccg tggagctcct
     2761 tcccgtgaat ggggaattca gcctggatga cctccagccg tggcaccctt ttggggtgga
     2821 ctctgtgcca gccaataccg aaaatgaagt cgagcctgtt gacgcccgcc ccgctgctga
     2881 ccgaggactg accactcgac caggttctgg gctgacaaac atcaagacgg aagagatctc
     2941 ggaagtgaag atggatgcag aattcggaca tgattcagga tttgaagtcc gccatcaaaa
     3001 actggtgttc tttgctgaag atgtgggttc gaacaaaggc gccatcatcg gactcatggt
     3061 gggcggcgtt gtcatagcaa ccgtgattgt catcaccctg gtgatgttga agaagaaaca
     3121 gtacacatcc atccatcatg gcgtggtgga ggtcgacgcc gccgtgaccc cagaggagcg
     3181 ccatctctcc aagatgcagc agaacggata tgagaatcca acttacaagt tctttgagca
     3241 aatgcagaac ctcgaggact acaaagacca tgacggtgat tataaagatc atgacatcga
     3301 ctacaaggat gacgatgaca agtagtgagg gcccgtttaa acccgctgat cagcctcgac
     3361 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     3421 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     3481 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     3541 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     3601 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     3661 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3721 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3781 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3841 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3901 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3961 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     4021 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     4081 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4141 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4201 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4261 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4321 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     4381 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     4441 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     4501 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     4561 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     4621 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     4681 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     4741 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     4801 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     4861 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     4921 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     4981 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     5041 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     5101 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     5161 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     5221 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     5281 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     5341 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     5401 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     5461 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5521 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     5581 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     5641 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     5701 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     5761 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5821 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5881 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5941 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6001 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6061 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6121 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6181 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6241 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6301 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6361 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6421 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     6481 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6541 gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6601 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6661 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6721 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6781 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6841 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6901 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6961 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7021 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7081 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7141 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7201 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7261 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7321 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7381 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7441 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7501 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7561 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7621 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7681 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7741 gaaaagtgcc acctgacgtc
//
