LOCUS       V030294                 7934 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030294
VERSION     V030294
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..3424
                     /label="PPP1R13L(NM_006663)"
                     /note="PPP1R13L(NM_006663)"
                     /gene="PPP1R13L"
     CDS             3431..3496
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3534..3758
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3804..4232
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4246..4575
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4642..5436
                     /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    5610..5731
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5780..5796)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5804..5820
                     /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(5828..5858)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5873..5894
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6182..6767)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6938..7798)
                     /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(7799..7903)
                     /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 atggacagcg aggcattcca
      961 gagcgcgcgg gactttctgg acatgaactt ccagtcgctg gccatgaaac acatggatct
     1021 gaagcagatg gagctggaca cggcggcggc caaggtggat gaactgacca agcagctgga
     1081 gtcgctgtgg tcagactctc ccgcgcctcc tggcccgcag gccggacccc cttctaggcc
     1141 gccccggtac agctccagct cgatccctga gcccttcggc agccgagggt ccccccggaa
     1201 ggcggccacc gacggcgcag acaccccgtt cggacgatca gagagtgccc caaccctaca
     1261 cccctacagc ccgctgtccc ccaagggacg gccgtcgtcg ccgcgcaccc cgctctacct
     1321 gcagccggac gcctacggca gcctggaccg cgcgacctcg ccccggcccc gcgccttcga
     1381 tggcgcaggc agctccctcg gccgtgcgcc ctccccgcgg cccgggccag gcccgctccg
     1441 ccagcagggt ccccccacgc ctttcgactt cctgggccgc gcaggctccc cccgcggcag
     1501 ccccctggcg gaggggcccc aggccttctt ccccgagcgt gggccgtcac cgcgcccccc
     1561 tgccacagcc tacgacgcgc cagcgtccgc cttcgggagc tccctgctag gctccggcgg
     1621 cagcgcattc gccccgcctc tgcgcgcgca agacgacctg acgctgcgcc ggcggcctcc
     1681 gaaagcctgg aacgagtctg acctggacgt ggcgtacgag aagaagcctt cgcagacagc
     1741 gagctatgaa cgcctggacg tcttcgcaag gcctgcctcg ccgagcctgc agctgttgcc
     1801 ttggagggag agcagcctgg atggactggg gggcaccggc aaggacaacc tcactagcgc
     1861 caccctgccg cgcaattaca aggtctctcc tctggccagc gaccggcgtt cagacgcggg
     1921 cagctaccgg cgctcgctgg gctccgcggg gccgtcgggc actttgcctc gcagctggca
     1981 gcccgtcagc cgcatcccca tgcccccctc cagcccccag ccccgcgggg ccccgcgcca
     2041 gcgtcccatc cccctcagca tgatcttcaa gctgcagaac gccttctggg agcacggggc
     2101 cagccgcgcc atgctccctg ggtcccccct cttcacccga gcacccccgc ctaagctgca
     2161 gccccaacca caaccacagc cccagccaca atcacaacca cagccccagc tgcccccaca
     2221 gccccagacc caaccccaaa cccctacccc agccccccag catccccaac agacatggcc
     2281 ccctgtgaac gaaggacccc ccaaaccccc caccgagctg gagcctgagc cggagataga
     2341 ggggctgctg acaccagtgc tggaggctgg cgatgtggat gaaggccctg tagcaaggcc
     2401 tctcagcccc acgaggctgc agccagcact gccaccggag gcacagtcgg tgcccgagct
     2461 ggaggaggtg gcacgggtgt tggcggaaat tccccggccc ctcaaacgca ggggctccat
     2521 ggagcaggcc cctgctgtgg ccctgccccc tacccacaag aaacagtacc agcagatcat
     2581 cagccgcctc ttccatcgtc atggggggcc agggcccggg gggccggagc cagagctgtc
     2641 ccccatcact gagggatctg aggccagggc agggccccct gctcctgccc caccagctcc
     2701 cattccaccc ccggccccgt cccagagcag cccaccagag cagccgcaga gcatggagat
     2761 gcgctctgtg ctgcggaagg cgggctcccc gcgcaaggcc cgccgcgcgc gcctcaaccc
     2821 tctggtgctc ctcctggacg cggcgctgac cggggagctg gaggtggtgc agcaggcggt
     2881 gaaggagatg aacgacccga gccagcccaa cgaggagggc atcactgcct tgcacaacgc
     2941 catctgcggc gccaactact ctatcgtgga tttcctcatc accgcgggtg ccaatgtcaa
     3001 ctcccccgac agccacggct ggacaccctt gcactgcgcg gcgtcgtgca acgacacagt
     3061 catctgcatg gcgctggtgc agcacggcgc tgcaatcttc gccaccacgc tcagcgacgg
     3121 cgccaccgcc ttcgagaagt gcgaccctta ccgcgagggt tatgctgact gcgccaccta
     3181 cctggcagac gtcgagcaga gtatggggct gatgaacagc ggggcagtgt acgctctctg
     3241 ggactacagc gccgagttcg gggacgagct gtccttccgc gagggcgagt cggtcaccgt
     3301 gctgcggagg gacgggccgg aggagaccga ctggtggtgg gccgcgctgc acggccagga
     3361 gggctacgtg ccgcggaact acttcgggct gttccccagg gtgaagcctc aaaggagtaa
     3421 agtcctcgag gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa
     3481 ggatgacgat gacaagtagt gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     3541 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3601 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3661 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3721 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     3781 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3841 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3901 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3961 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     4021 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4081 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4141 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     4201 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     4261 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4321 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4381 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4441 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4501 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4561 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     4621 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     4681 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4741 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4801 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     4861 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4921 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4981 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     5041 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     5101 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     5161 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     5221 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     5281 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     5341 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     5401 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     5461 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     5521 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     5581 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     5641 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     5701 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     5761 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     5821 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     5881 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     5941 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     6001 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     6061 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     6121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     6421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     6661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt
     6721 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6781 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6841 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6901 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6961 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     7021 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     7081 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     7141 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     7201 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     7261 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     7321 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     7381 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     7441 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     7501 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     7561 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     7621 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     7681 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     7741 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     7801 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7861 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     7921 tgccacctga cgtc
//
