LOCUS       V036959                 7297 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036959
VERSION     V036959
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             964..2787
                     /label="PLCZ1(NM_033123)"
                     /note="PLCZ1(NM_033123)"
                     /gene="PLCZ1"
     CDS             2794..2859
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2897..3121
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3167..3595
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3609..3938
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4005..4799
                     /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    4973..5094
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5143..5159)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5167..5183
                     /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(5191..5221)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5236..5257
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5545..6130)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6301..7161)
                     /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(7162..7266)
                     /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 atccactagt ccagtgtggt ggaattcgcc
      961 accatggaaa tgagatggtt tttgtcaaag attcaggatg acttcagagg tggaaaaatt
     1021 aacctagaaa aaactcagag gttacttgaa aaattagata ttcggtgcag ttatattcat
     1081 gtgaaacaga tttttaagga caatgacagg ctgaaacaag gaagaatcac catagaagaa
     1141 tttagagcaa tttatcgaat tatcacgcac agagaagaaa ttattgagat tttcaacaca
     1201 tattctgaaa accggaaaat tcttttagca agtaatctgg ctcaatttct gacacaagaa
     1261 caatatgcag ctgagatgag taaagctatt gcttttgaga tcattcagaa atacgagcct
     1321 atcgaagaag ttaggaaagc acaccaaatg tcattagaag gttttacaag atacatggat
     1381 tcacgtgaat gtctactgtt taaaaatgaa tgtagaaaag tttatcaaga tatgactcat
     1441 ccattaaatg attattttat ttcatcttca cataacacat atttggtatc tgatcaatta
     1501 ttgggaccaa gtgacctttg gggatatgta agtgcccttg tgaaaggatg ccgttgtttg
     1561 gagattgact gctgggatgg agcacaaaat gaacctgttg tatatcatgg ctacacactc
     1621 acaagcaaac ttctgtttaa aactgttatc caagctatac acaagtatgc attcatgaca
     1681 tctgactacc cagtggtgct ctctttagaa aatcactgct ccactgccca acaagaagta
     1741 atggcagaca atttgcaggc tacttttgga gagtccttgc tttctgatat gcttgatgat
     1801 tttcctgata ctctaccatc accagaggca ctaaaattca aaatattagt taaaaataag
     1861 aaaataggaa ccttaaagga aacccatgaa agaaaaggtt ctgataagcg tggagacaat
     1921 caagacaagg aaacaggggt aaaaaagtta cctggagtaa tgcttttcaa gaaaaagaag
     1981 accaggaagc taaaaattgc tctggcctta tctgatcttg tcatttatac gaaagctgag
     2041 aaattcaaaa gctttcaaca ttcaagatta tatcagcaat ttaatgaaaa taattctatt
     2101 ggggagacac aagcccgaaa actttcaaaa ttgcgagtcc atgagtttat ttttcacacc
     2161 aggaagttca ttaccagaat atatcccaaa gcaacaagag cagactcttc taattttaat
     2221 ccccaagaat tttggaatat aggttgtcaa atggtggctt taaatttcca gacccctggt
     2281 ctgcccatgg atctgcaaaa tgggaaattt ttggataatg gtggttctgg atatattttg
     2341 aaaccacatt tcttaagaga gagtaaatca tactttaacc caagtaacat aaaagagggt
     2401 atgccaatta cacttacaat aaggctcatc agtggtatcc agttgcctct tactcattca
     2461 tcatctaaca aaggtgattc attagtaatt atagaagttt ttggtgttcc aaatgatcaa
     2521 atgaagcagc agactcgtgt aattaaaaaa aatgctttta gtccaagatg gaatgaaaca
     2581 ttcacattta ttattcatgt cccagaattg gcattgatac gttttgttgt tgaaggtcaa
     2641 ggtttaatag caggaaatga atttcttggg caatatactt tgccacttct atgcatgaac
     2701 aaaggttatc gtcgtattcc tctgttttcc agaatgggtg agagccttga gcctgcttca
     2761 ctgtttgttt atgtttggta cgtcagactc gaggactaca aagaccatga cggtgattat
     2821 aaagatcatg acatcgacta caaggatgac gatgacaagt agtgagggcc cgtttaaacc
     2881 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     2941 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     3001 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     3061 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     3121 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     3181 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3241 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3301 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3361 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     3421 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3481 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     3541 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     3601 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3661 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3721 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3781 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3841 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     3901 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     3961 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     4021 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4081 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     4141 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     4201 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4261 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4321 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4381 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4441 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4501 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     4561 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4621 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4681 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4741 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     4801 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     4861 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4921 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     4981 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     5041 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     5101 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     5161 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     5221 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     5281 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5341 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5401 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5461 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5521 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5581 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5641 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5701 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5761 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5821 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5881 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5941 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     6001 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6061 acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6121 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6181 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6241 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6301 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6361 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6421 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6481 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6541 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6601 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6661 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6721 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6781 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6841 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6901 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6961 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7021 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7081 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7141 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7201 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7261 tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
