LOCUS       V033732                 8736 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033732
VERSION     V033732
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..3416
                     /label="Pik3c3(NM_181414)"
                     /note="Pik3c3(NM_181414)"
                     /gene="Pik3c3"
     CDS             3423..3488
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3512..4100
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4166..4214
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4416..4844
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4858..5187
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5254..6048
                     /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    6224..6345
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6394..6410)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6418..6434
                     /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(6442..6472)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6487..6508
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6796..7384)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7555..8415)
                     /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(8416..8520)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggg ggaggcggag aagttccact
      781 acatctacag ctgtgacctg gacatcaacg tgcagctgaa gatagggagc ttggaaggga
      841 agagagaaca aaagagctat aaagctgtcc tagaagatcc catgttaaag ttttctgggc
      901 tataccaaga gacatgctcc gacctctatg tgacttgtca agtgtttgct gaagggaagc
      961 ccctggcctt accagtcaga acatcctaca aggcgtttag tacaagatgg aattggaacg
     1021 agtggctgaa acttccggtg aagtaccctg acctgcccag gaatgcccaa gtggccttga
     1081 ctatatggga tgtgtatgga ccgggaagcg ctgtgccagt gggaggaaca accgtgtcgc
     1141 tctttggaaa atatggcatg tttcgccaag gaatgcatga cttgaaggtc tggcctaacg
     1201 tggaggcaga tggttctgaa cctacaagaa ctcccggcag aacaagcagc acactgtcag
     1261 aagatcagat gagccgcctc gccaagctca ccaaggctca tcggcaagga cacatggtga
     1321 aagtggattg gctggacaga ttaacattta gagagataga aatgataaat gagagtgaga
     1381 aacgaagctc taatttcatg tacttgatgg ttgagtttcg ctgtgtcaag tgcgatgaca
     1441 aggagtatgg cattgtttac tatgaaaagg atggtgacga atcatctcca attttaacca
     1501 gttttgagtt agtgaaagtt cctgatcctc aaatgtctat ggagaattta gtggagagca
     1561 aacaccacaa gcttgctcgg agcttaagaa gcgggccatc tgaccacgat ctcaaaccca
     1621 acgctaccac aagagatcag ctaaatatta ttgtgagtta cccaccaacc aagcaactca
     1681 catatgaaga acaagatctt gtttggaaat ttagatatta tcttactaat caagaaaaag
     1741 ctctgacgaa gttcttgaag tgtgttaatt gggacctgcc ccaggaggcc aagcaggccc
     1801 tggaacttct gggaaagtgg aagccaatgg atgtagagga ctccctggag ctgctctcct
     1861 ctcattacac caaccccacc gtgaggcgct acgctgtggc ccgcctgagg caggccgatg
     1921 atgaggattt gctgatgtac ttactgcagc tggtccaggc tctgaaatat gaaaactttg
     1981 atgacataaa gaatggtttg gaacctacca agaaagatag tcaaacttca gcatcagaaa
     2041 gtctgtcaaa ttctggagtc agttctggag acatagatag ctcccaaatt ataaccaacc
     2101 ctcttcctcc cgtggcctcc cctcctcctg catctaaagc aaaggaagtt tccgatgggg
     2161 aaaatctcga gcaagatcta tgtacgttct tgatatcaag agcctgtaag aactcaacac
     2221 tggctaatta tttatactgg tatgtgatag tggagtgtga agatcaggac acccagcagc
     2281 gggacccaaa gactcacgag atgtacctga acgtgatgag gcgcttcagc caagccttgc
     2341 tcaagggtga caagtctgtg agagtcatgc gctccctgct ggcggctcag cagacctttg
     2401 tagatcgcct ggtccatctg atgaaggcgg tgcagagaga aagtgggaac cgcaagaaga
     2461 agaatgagag acttcaggcc ttgcttggag ataatgagaa aatgaactta tcagatgtgg
     2521 aactgatccc attgccgctg gagccacagg tgaaaataag gggcatcatc ccggaaacag
     2581 ctaccctgtt caagagtgct cttatgcctg cccagctgtt cttcaagact gaagatggag
     2641 gcaaataccc agttattttc aagcatggag acgacttgcg tcaagatcag cttattcttc
     2701 agatcatctc cctcatggac aagctgttac ggaaagaaaa cctggatttg aaattgaccc
     2761 catacaaggt gttagccact agcacaaaac atggcttcat gcagttcatc cagtcggttc
     2821 ctgtcgcaga agttcttgac acagagggaa gcattcagaa ctttttcagg aaatatgccc
     2881 ccagtgagac tggaccctat ggcatcagtg cagaggtcat ggacacttac gttaaaagct
     2941 gtgctggata ctgtgtgatt acgtacattc ttggagttgg agaccggcac ctggataacc
     3001 ttttgttaac aaagacaggc aaactcttcc atatagattt cgggtatatt ttgggtcgag
     3061 atccgaagcc cctccctcct ccgatgaagc tgaacaagga gatggtggaa gggatgggtg
     3121 gcacccagag tgagcagtac caagagttcc gaaagcagtg ctacacagcc ttcctccacc
     3181 tgcgaaggta ttccaatctg atcttgaact tgttctcctt gatggttgat gcaaacattc
     3241 cagatattgc tcttgaacca gataaaactg taaaaaaggt tcaggataag ttccgcctgg
     3301 acctgtcaga tgaggaggct gtgcactaca tgcagagtct gatcgacgag agcgtccacg
     3361 cactgttcgc cgccgtggtg gagcagatcc ataagtttgc ccagtactgg agaaaaggat
     3421 ccgactacaa agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg
     3481 atgacaagta gtgagggccc gatatctcga caatcaacct ctggattaca aaatttgtga
     3541 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     3601 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     3661 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     3721 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     3781 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     3841 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     3901 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     3961 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     4021 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     4081 cctttgggcc gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg gaaggagaca
     4141 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt
     4201 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     4261 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     4321 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     4381 agatctgcgc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     4441 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     4501 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4561 tctaaatcgg ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4621 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4681 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4741 actcaaccct atctcggtct attcttttga tttataaggg attttgggga tttcggccta
     4801 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     4861 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4921 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     4981 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     5041 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     5101 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     5161 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     5221 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     5281 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     5341 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     5401 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     5461 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     5521 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     5581 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     5641 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     5701 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     5761 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     5821 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     5881 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     5941 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     6001 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     6061 ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     6121 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     6181 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     6241 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     6301 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     6361 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     6421 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     6481 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     6541 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     6601 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     6661 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     6721 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6781 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6841 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6901 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6961 tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt
     7021 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     7081 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     7141 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     7201 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     7261 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     7321 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     7381 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     7441 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     7501 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     7561 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     7621 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     7681 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     7741 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     7801 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     7861 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     7921 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7981 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     8041 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     8101 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     8161 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     8221 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     8281 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     8341 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     8401 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     8461 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     8521 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     8581 tggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     8641 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     8701 ggcttgaccg acaattgcat gaagaatctg cttagg
//
