LOCUS       V043192                 8259 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043192
VERSION     V043192
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..2939
                     /label="PIK3R2(NM_005027)"
                     /note="PIK3R2(NM_005027)"
                     /gene="PIK3R2"
     CDS             2946..3011
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3035..3623
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3689..3737
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3939..4367
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4381..4710
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4777..5571
                     /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    5747..5868
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5917..5933)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5941..5957
                     /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(5965..5995)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6010..6031
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6319..6907)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7078..7938)
                     /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(7939..8043)
                     /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 ccaccatggc gggccctgag ggcttccagt
      781 accgcgctct gtacccgttc cgccgggagc ggccggagga cctggagctg ctgcccggcg
      841 acgtgctggt agtgagccgg gcggccttgc aggcgctggg cgtggccgag ggtggcgagc
      901 gctgcccaca gagcgtgggc tggatgcccg gcctcaacga gcgcacacgg cagcgaggtg
      961 acttccctgg cacctatgtg gagttcctgg ggcccgtggc cctggcccgg cccggccctc
     1021 gcccacgggg cccccgccca ctgcccgcca ggccccgtga tggggcccct gagccaggcc
     1081 tcacactccc cgacttgccc gagcagttct ccccacctga tgtggctccc cctcttctgg
     1141 tgaagcttgt ggaggccatt gaaaggacag ggctggacag cgaatctcac taccgcccgg
     1201 agctgcccgc accgcgtaca gactggtccc tgagcgacgt ggatcagtgg gacacggcag
     1261 ccctggctga cggcattaag agcttcctgc tggcactgcc cgcgccgctc gtgacccccg
     1321 aggcctcggc cgaggcgcgc cgggccctgc gggaggccgc ggggcccgtg gggccggcgc
     1381 tggagccacc gacgctgccg ctgcaccgcg cgctcacgct gcgcttcctg ctccagcacc
     1441 tgggccgcgt ggccagccgc gccccggccc tgggtcccgc ggtccgggcc ctgggcgcca
     1501 cctttgggcc gctgctgctg cgcgcgccgc cgccgccgtc ctcgccgccg ccagggggcg
     1561 ctcccgacgg gagtgagccc agccctgact tcccggcgct gctggtggag aagctgcttc
     1621 aggaacactt ggaagagcag gaggttgcgc ccccagcgct gccgcctaaa ccccccaagg
     1681 caaagccggc ctccacagtc ctggccaatg gagggagccc accctccctg caggatgctg
     1741 agtggtactg gggggacatt tcaagggagg aggtgaacga gaaactccgg gacactcccg
     1801 atggcacctt cctagtccga gatgcttcta gcaagatcca gggcgagtac acgctgaccc
     1861 tcaggaaagg cgggaacaat aagctgatca aggtcttcca ccgagatggg cactatggct
     1921 tctcagagcc actcaccttc tgctccgttg tggacctcat caatcactac cgccacgagt
     1981 ctctggccca gtacaatgcc aagctggaca cacggctcct ctaccctgtg tccaaatacc
     2041 agcaggacca gattgtcaag gaggacagcg tggaggcagt gggcgcccag cttaaggtct
     2101 atcaccagca gtaccaggac aagagccgcg agtatgacca gctttatgaa gagtacacac
     2161 ggacctccca ggagctgcag atgaagcgta ctgcaattga ggccttcaat gagactatca
     2221 agatctttga agagcagggc cagactcaag agaaatgcag caaggaatac ctggagcgct
     2281 tccggcgtga gggcaacgag aaagagatgc aaaggatcct gctgaactcc gagcggctca
     2341 agtcccgcat tgccgagatc catgagagcc gcacgaagct ggagcagcag ctgcgggccc
     2401 aggcctcgga caacagagag atcgacaagc gcatgaacag cctcaagccg gacctcatgc
     2461 agctgcgcaa gatccgagac cagtacctcg tgtggctcac ccagaaaggc gcccggcaga
     2521 agaaaatcaa cgagtggctg gggattaaaa atgagactga ggaccagtac gcactcatgg
     2581 aggacgagga cgatctcccg caccacgagg aacgcacttg gtacgtgggc aagatcaacc
     2641 gcacgcaggc agaggagatg ctgagtggca agcgggatgg caccttcctc atccgcgaga
     2701 gcagccagcg gggctgctac gcctgctccg tggtagtgga cggcgacacc aagcactgcg
     2761 tcatctaccg cacggccacc ggcttcggct tcgcggagcc ctacaacctg tacgggtcgc
     2821 tgaaggagct ggtgctgcac taccagcacg cctcgctggt gcagcacaac gacgcgctca
     2881 ccgtcaccct ggcgcaccca gtgcgcgccc cgggccccgg cccgccgcct gccgcccgcg
     2941 gatccgacta caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg
     3001 acgatgacaa gtagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg
     3061 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     3121 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     3181 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     3241 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     3301 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     3361 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     3421 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     3481 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     3541 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     3601 ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag
     3661 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg
     3721 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     3781 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     3841 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     3901 agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     3961 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     4021 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4081 agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4141 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4201 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4261 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc
     4321 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     4381 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4441 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     4501 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4561 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4621 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     4681 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     4741 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4801 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4861 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4921 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     4981 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     5041 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     5101 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     5161 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     5221 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     5281 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     5341 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5401 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5461 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     5521 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5581 tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5641 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5701 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     5761 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5821 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     5881 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5941 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     6001 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     6061 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     6121 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6181 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6241 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6301 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6361 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6421 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6481 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     6541 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6601 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6661 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6721 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     6781 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6841 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6901 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6961 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     7021 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     7081 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7141 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7201 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7261 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7321 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7381 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7441 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7501 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7561 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7621 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7681 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7741 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7801 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7861 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7921 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7981 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     8041 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     8101 ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     8161 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     8221 caaggcttga ccgacaattg catgaagaat ctgcttagg
//
