LOCUS       V041134                 7802 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041134
VERSION     V041134
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             683..2449
                     /label="PPP2R1A(NM_014225)"
                     /note="PPP2R1A(NM_014225)"
                     /gene="PPP2R1A"
     CDS             2489..2554
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2578..3166
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3232..3280
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3482..3910
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3924..4253
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4320..5114
                     /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    5290..5411
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5460..5476)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5484..5500
                     /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(5508..5538)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5553..5574
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5862..6450)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6621..7481)
                     /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(7482..7586)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatggcggc ggccgacggc gacgactcgc tgtaccccat
      721 cgcggtgctc atagacgaac tccgcaatga ggacgttcag cttcgcctca acagcatcaa
      781 gaagctgtcc accatcgcct tggcccttgg ggttgaaagg acccgaagtg agcttctgcc
      841 tttccttaca gataccatct atgatgaaga tgaggtcctc ctggccctgg cagaacagct
      901 gggaaccttc actaccctgg tgggaggccc agagtacgtg cactgcctgc tgccaccgct
      961 ggagtcgctg gccacagtgg aggagacagt ggtgcgggac aaggcagtgg agtccttacg
     1021 ggccatctca cacgagcact cgccctctga cctggaggcg cactttgtgc cgctagtgaa
     1081 gcggctggcg ggcggcgact ggttcacctc ccgcacctcg gcctgcggcc tcttctccgt
     1141 ctgctacccc cgagtgtcca gtgctgtgaa ggcggaactt cgacagtact tccggaacct
     1201 gtgctcagat gacaccccca tggtgcggcg ggccgcagcc tccaagctgg gggagtttgc
     1261 caaggtgctg gagctggaca acgtcaagag tgagatcatc cccatgttct ccaacctggc
     1321 ctctgacgag caggactcgg tgcggctgct ggcggtggag gcgtgcgtga acatcgccca
     1381 gcttctgccc caggaggatc tggaggccct ggtgatgccc actctgcgcc aggccgctga
     1441 agacaagtcc tggcgcgtcc gctacatggt ggctgacaag ttcacagagc tccagaaagc
     1501 agtggggcct gagatcacca agacagacct ggtccctgcc ttccagaacc tgatgaaaga
     1561 ctgtgaggcc gaggtgaggg ccgcagcctc ccacaaggtc aaagagttct gtgaaaacct
     1621 ctcagctgac tgtcgggaga atgtgatcat gtcccagatc ttgccctgca tcaaggagct
     1681 ggtgtccgat gccaaccaac atgtcaagtc tgccctggcc tcagtcatca tgggtctctc
     1741 tcccatcttg ggcaaagaca acaccatcga gcacctcttg cccctcttcc tggctcagct
     1801 gaaggatgag tgccctgagg tacggctgaa catcatctct aacctggact gtgtgaacga
     1861 ggtgattggc atccggcagc tgtcccagtc cctgctccct gccattgtgg agctggctga
     1921 ggacgccaag tggcgggtgc ggctggccat cattgagtac atgcccctcc tggctggaca
     1981 gctgggagtg gagttctttg atgagaaact taactccttg tgcatggcct ggcttgtgga
     2041 tcatgtatat gccatccgcg aggcagccac cagcaacctg aagaagctag tggaaaagtt
     2101 tgggaaggag tgggcccatg ccacaatcat ccccaaggtc ttggccatgt ccggagaccc
     2161 caactacctg caccgcatga ctacgctctt ctgcatcaat gtgctgtctg aggtctgtgg
     2221 gcaggacatc accaccaagc acatgctacc cacggttctg cgcatggctg gggacccggt
     2281 tgccaatgtc cgcttcaatg tggccaagtc tctgcagaag atagggccca tcctggacaa
     2341 cagcaccttg cagagtgaag tcaagcccat cctagagaag ctgacccagg accaggatgt
     2401 ggacgtcaaa tactttgccc aggaggctct gactgttctg tctctcgccg aattctgcag
     2461 atatccagca cagtggcggc cgctcgagga ctacaaagac catgacggtg attataaaga
     2521 tcatgacatc gactacaagg atgacgatga caagtagtga gggcccgata tctcgacaat
     2581 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     2641 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     2701 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     2761 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     2821 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     2881 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     2941 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc
     3001 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     3061 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     3121 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg aaacggggga
     3181 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     3241 aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     3301 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     3361 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     3421 gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag ggggtatccc
     3481 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3541 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3601 acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg gttccgattt
     3661 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3721 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3781 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     3841 taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3901 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3961 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4021 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4081 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4141 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     4201 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     4261 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     4321 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     4381 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4441 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     4501 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4561 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4621 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     4681 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4741 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4801 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     4861 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     4921 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     4981 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     5041 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     5101 acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac
     5161 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     5221 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     5281 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     5341 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     5401 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     5461 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     5521 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     5581 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     5641 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     5701 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5761 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5821 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5881 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5941 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6001 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc
     6061 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6121 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6181 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6241 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6301 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6361 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6421 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6481 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6541 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6601 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     6661 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     6721 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     6781 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6841 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6901 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6961 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     7021 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     7081 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     7141 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     7201 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     7261 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     7321 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     7381 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7441 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7501 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7561 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     7621 ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc tctgatgccg
     7681 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
     7741 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
     7801 gg
//
