LOCUS       V036782                 8435 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036782
VERSION     V036782
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             884..1591
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
                     /label="mCherry"
     CDS             1628..3937
                     /label="APP(NM_000484)"
                     /note="APP(NM_000484)"
                     /gene="APP"
     CDS             3938..3955
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     polyA_signal    4002..4226
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4272..4700
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4714..5043
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5110..5904
                     /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    6078..6199
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6248..6264)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6272..6288
                     /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(6296..6326)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6341..6362
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6650..7235)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7406..8266)
                     /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(8267..8371)
                     /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 ggctagcgct accggtcgcc accatggtga gcaagggcga
      901 ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc
      961 cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
     1021 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct
     1081 gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga
     1141 ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
     1201 cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa
     1261 ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat
     1321 gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat
     1381 caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta
     1441 caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat
     1501 cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca
     1561 ctccaccggc ggcatggacg agctgtacaa gggtggaggc ggatcaggag gcggtggatc
     1621 tctcgagatg ctgcccggtt tggcactgct cctgctggcc gcctggacgg ctcgggcgct
     1681 ggaggtaccc actgatggta atgctggcct gctggctgaa ccccagattg ccatgttctg
     1741 tggcagactg aacatgcaca tgaatgtcca gaatgggaag tgggattcag atccatcagg
     1801 gaccaaaacc tgcattgata ccaaggaagg catcctgcag tattgccaag aagtctaccc
     1861 tgaactgcag atcaccaatg tggtagaagc caaccaacca gtgaccatcc agaactggtg
     1921 caagcggggc cgcaagcagt gcaagaccca tccccacttt gtgattccct accgctgctt
     1981 agttggtgag tttgtaagtg atgcccttct cgttcctgac aagtgcaaat tcttacacca
     2041 ggagaggatg gatgtttgcg aaactcatct tcactggcac accgtcgcca aagagacatg
     2101 cagtgagaag agtaccaact tgcatgacta cggcatgttg ctgccctgcg gaattgacaa
     2161 gttccgaggg gtagagtttg tgtgttgccc actggctgaa gaaagtgaca atgtggattc
     2221 tgctgatgcg gaggaggatg actcggatgt ctggtggggc ggagcagaca cagactatgc
     2281 agatgggagt gaagacaaag tagtagaagt agcagaggag gaagaagtgg ctgaggtgga
     2341 agaagaagaa gccgatgatg acgaggacga tgaggatggt gatgaggtag aggaagaggc
     2401 tgaggaaccc tacgaagaag ccacagagag aaccaccagc attgccacca ccaccaccac
     2461 caccacagag tctgtggaag aggtggttcg agaggtgtgc tctgaacaag ccgagacggg
     2521 gccgtgccga gcaatgatct cccgctggta ctttgatgtg actgaaggga agtgtgcccc
     2581 attcttttac ggcggatgtg gcggcaaccg gaacaacttt gacacagaag agtactgcat
     2641 ggccgtgtgt ggcagcgcca tgtcccaaag tttactcaag actacccagg aacctcttgc
     2701 ccgagatcct gttaaacttc ctacaacagc agccagtacc cctgatgccg ttgacaagta
     2761 tctcgagaca cctggggatg agaatgaaca tgcccatttc cagaaagcca aagagaggct
     2821 tgaggccaag caccgagaga gaatgtccca ggtcatgaga gaatgggaag aggcagaacg
     2881 tcaagcaaag aacttgccta aagctgataa gaaggcagtt atccagcatt tccaggagaa
     2941 agtggaatct ttggaacagg aagcagccaa cgagagacag cagctggtgg agacacacat
     3001 ggccagagtg gaagccatgc tcaatgaccg ccgccgcctg gccctggaga actacatcac
     3061 cgctctgcag gctgttcctc ctcggcctcg tcacgtgttc aatatgctaa agaagtatgt
     3121 ccgcgcagaa cagaaggaca gacagcacac cctaaagcat ttcgagcatg tgcgcatggt
     3181 ggatcccaag aaagccgctc agatccggtc ccaggttatg acacacctcc gtgtgattta
     3241 tgagcgcatg aatcagtctc tctccctgct ctacaacgtg cctgcagtgg ccgaggagat
     3301 tcaggatgaa gttgatgagc tgcttcagaa agagcaaaac tattcagatg acgtcttggc
     3361 caacatgatt agtgaaccaa ggatcagtta cggaaacgat gctctcatgc catctttgac
     3421 cgaaacgaaa accaccgtgg agctccttcc cgtgaatgga gagttcagcc tggacgatct
     3481 ccagccgtgg cattcttttg gggctgactc tgtgccagcc aacacagaaa acgaagttga
     3541 gcctgttgat gcccgccctg ctgccgaccg aggactgacc actcgaccag gttctgggtt
     3601 gacaaatatc aagacggagg agatctctga agtgaagatg gatgcagaat tccgacatga
     3661 ctcaggatat gaagttcatc atcaaaaatt ggtgttcttt gcagaagatg tgggttcaaa
     3721 caaaggtgca atcattggac tcatggtggg cggtgttgtc atagcgacag tgatcgtcat
     3781 caccttggtg atgctgaaga agaaacagta cacatccatt catcatggtg tggtggaggt
     3841 tgacgccgct gtcaccccag aggagcgcca cctgtccaag atgcagcaga acggctacga
     3901 aaatccaacc tacaagttct ttgagcagat gcagaaccat caccatcacc atcactaggg
     3961 atcctctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     4021 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     4081 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     4141 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     4201 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     4261 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4321 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4381 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4441 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     4501 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4561 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     4621 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     4681 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4741 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4801 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4861 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4921 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4981 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     5041 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     5101 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     5161 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     5221 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     5281 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     5341 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     5401 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     5461 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     5521 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     5581 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     5641 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     5701 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     5761 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5821 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5881 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     5941 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     6001 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     6061 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     6121 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     6181 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     6241 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     6301 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     6361 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     6421 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     6481 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     6541 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     6601 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6661 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6721 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6781 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6841 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6901 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6961 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     7021 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     7081 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     7141 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca
     7201 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     7261 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     7321 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     7381 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     7441 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     7501 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     7561 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     7621 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     7681 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     7741 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     7801 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     7861 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7921 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7981 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     8041 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     8101 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     8161 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     8221 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     8281 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     8341 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     8401 tcgacggatc gggagatctc ccgatcccct atggt
//
