LOCUS       V020700                 8174 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020700
VERSION     V020700
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..2854
                     /label="PIK3R1(NM_181523)"
                     /note="PIK3R1(NM_181523)"
                     /gene="PIK3R1"
     CDS             2861..2926
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2950..3538
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3604..3652
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3854..4282
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4296..4625
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4692..5486
                     /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    5662..5783
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5832..5848)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5856..5872
                     /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(5880..5910)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5925..5946
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6234..6822)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6993..7853)
                     /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(7854..7958)
                     /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 ccatgagtgc tgaggggtac cagtacagag cgctgtatga
      721 ttataaaaag gaaagagaag aagatattga cttgcacttg ggtgacatat tgactgtgaa
      781 taaagggtcc ttagtagctc ttggattcag tgatggacag gaagccaggc ctgaagaaat
      841 tggctggtta aatggctata atgaaaccac aggggaaagg ggggactttc cgggaactta
      901 cgtagaatat attggaagga aaaaaatctc gcctcccaca ccaaagcccc ggccacctcg
      961 gcctcttcct gttgcaccag gttcttcgaa aactgaagca gatgttgaac aacaagcttt
     1021 gactctcccg gatcttgcag agcagtttgc ccctcctgac attgccccgc ctcttcttat
     1081 caagctcgtg gaagccattg aaaagaaagg tctggaatgt tcaactctat acagaacaca
     1141 gagctccagc aacctggcag aattacgaca gcttcttgat tgtgatacac cctccgtgga
     1201 cttggaaatg atcgatgtgc acgttttggc tgacgctttc aaacgctatc tcctggactt
     1261 accaaatcct gtcattccag cagccgttta cagtgaaatg atttctttag ctccagaagt
     1321 acaaagctcc gaagaatata ttcagctatt gaagaagctt attaggtcgc ctagcatacc
     1381 tcatcagtat tggcttacgc ttcagtattt gttaaaacat ttcttcaagc tctctcaaac
     1441 ctccagcaaa aatctgttga atgcaagagt actctctgaa attttcagcc ctatgctttt
     1501 cagattctca gcagccagct ctgataatac tgaaaacctc ataaaagtta tagaaatttt
     1561 aatctcaact gaatggaatg aacgacagcc tgcaccagca ctgcctccta aaccaccaaa
     1621 acctactact gtagccaaca acggtatgaa taacaatatg tccttacaag atgctgaatg
     1681 gtactgggga gatatctcga gggaagaagt gaatgaaaaa cttcgagata cagcagacgg
     1741 gacctttttg gtacgagatg cgtctactaa aatgcatggt gattatactc ttacactaag
     1801 gaaaggggga aataacaaat taatcaaaat atttcatcga gatgggaaat atggcttctc
     1861 tgacccatta accttcagtt ctgtggttga attaataaac cactaccgga atgaatctct
     1921 agctcagtat aatcccaaat tggatgtgaa attactttat ccagtatcca aataccaaca
     1981 ggatcaagtt gtcaaagaag ataatattga agctgtaggg aaaaaattac atgaatataa
     2041 cactcagttt caagaaaaaa gtcgagaata tgatagatta tatgaagaat atacccgcac
     2101 atcccaggaa atccaaatga aaaggacagc tattgaagca tttaatgaaa ccataaaaat
     2161 atttgaagaa cagtgccaga cccaagagcg gtacagcaaa gaatacatag aaaagtttaa
     2221 acgtgaaggc aatgagaaag aaatacaaag gattatgcat aattatgata agttgaagtc
     2281 tcgaatcagt gaaattattg acagtagaag aagattggaa gaagacttga agaagcaggc
     2341 agctgagtat cgagaaattg acaaacgtat gaacagcatt aaaccagacc ttatccagct
     2401 gagaaagacg agagaccaat acttgatgtg gttgactcaa aaaggtgttc ggcaaaagaa
     2461 gttgaacgag tggttgggca atgaaaacac tgaagaccaa tattcactgg tggaagatga
     2521 tgaagatttg ccccatcatg atgagaagac atggaatgtt ggaagcagca accgaaacaa
     2581 agctgaaaac ctgttgcgag ggaagcgaga tggcactttt cttgtccggg agagcagtaa
     2641 acagggctgc tatgcctgct ctgtagtggt ggacggcgaa gtaaagcatt gtgtcataaa
     2701 caaaacagca actggctatg gctttgccga gccctataac ttgtacagct ctctgaaaga
     2761 actggtgcta cattaccaac acacctccct tgtgcagcac aacgactccc tcaatgtcac
     2821 actagcctac ccagtatatg cacagcagag gcgactcgag gactacaaag accatgacgg
     2881 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccga
     2941 tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     3001 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     3061 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     3121 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     3181 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     3241 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     3301 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
     3361 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     3421 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     3481 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     3541 ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     3601 tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc
     3661 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     3721 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     3781 ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct
     3841 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3901 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3961 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg catcccttta
     4021 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     4081 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4141 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     4201 tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa tgagctgatt
     4261 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4321 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4381 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4441 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4501 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4561 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4621 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     4681 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4741 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4801 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4861 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4921 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4981 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     5041 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     5101 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     5161 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     5221 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     5281 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     5341 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     5401 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     5461 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa
     5521 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     5581 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     5641 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     5701 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     5761 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     5821 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5881 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5941 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     6001 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     6061 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     6121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     6181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     6241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     6301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     6361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     6421 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     6481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     6541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     6601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     6661 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     6721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6901 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6961 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     7021 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     7081 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     7141 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     7201 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     7261 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     7321 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     7381 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     7441 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     7501 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     7561 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     7621 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7681 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7741 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7801 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7861 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7921 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7981 cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc agtacaatct
     8041 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     8101 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     8161 agaatctgct tagg
//
