LOCUS       V020373                10076 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020373
VERSION     V020373
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..4756
                     /label="PIK3R4(NM_014602)"
                     /note="PIK3R4(NM_014602)"
                     /gene="PIK3R4"
     CDS             4763..4828
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4852..5440
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5506..5554
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5756..6184
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6198..6527
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6594..7388
                     /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    7564..7685
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7734..7750)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7758..7774
                     /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(7782..7812)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7827..7848
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8136..8724)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8895..9755)
                     /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(9756..9860)
                     /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 ccatgggaaa tcagcttgct ggcattgctc cctcccagat
      721 cctttctgta gagagttatt tttcagatat tcatgacttt gaatatgata aaagcctggg
      781 gagtactcgg ttttttaaag ttgctcgagc caagcaccga gaaggcctgg tcgttgtgaa
      841 ggtttttgca attcaggatc ccacattgcc tttaaccagc tataaacaag agctggagga
      901 actgaaaatc aggcttaatt ctgcacagaa ttgtctacct ttccagaaag catcagaaaa
      961 agcatctgag aaagcagcta tgctctttag gcagtatgtg cgagacaatc tctatgatcg
     1021 catcagtacc cgtccattct tgaataacat tgagaagcgc tggattgctt tccagatcct
     1081 gacagctgtg gaccaagcac acaaatctgg agttcgtcat ggggacatca agactgagaa
     1141 tgtgatggtc accagttgga attgggttct tctaactgat tttgccagtt ttaagcccac
     1201 ttatcttcca gaagacaacc cggcagattt caattatttc tttgacacat cacggaggag
     1261 aacttgctat attgctcctg aacgttttgt tgatggtggg atgtttgcca ctgagttaga
     1321 atatatgaga gatccttcaa ctccgcttgt agacttaaat agcaatcaga gaacaagagg
     1381 agagttgaag agagcaatgg acatcttttc agcaggttgt gtgatagctg agctttttac
     1441 agaaggtgta ccattatttg atctctctca acttttggct tatagaaatg gacatttttt
     1501 ccctgaacaa gtgctaaata aaattgaaga tcacagtatc agagaattgg taactcagat
     1561 gattcaccgt gagccagata aacgtttaga ggcagaagat tacttaaaac agcagcgtgg
     1621 caatgccttt cctgaaatat tttacacttt tcttcagccc tacatggccc agtttgccaa
     1681 ggaaacgttt ctttctgcag atgagcgtat tctggttata cggaaggatt tgggcaacat
     1741 tattcacaat ctctgtggac atgatctgcc agaaaaagcc gaaggagagc ctaaggaaaa
     1801 tgggctggtt atcttggtat ctgttataac atcctgccta cagaccctta aatactgtga
     1861 ttccaaacta gctgctttgg aactgattct tcatttggct ccaagattaa gtgttgaaat
     1921 ccttttggat cgtattactc catatctttt gcatttcagc aatgactctg ttcctagggt
     1981 gagggctgaa gccttgagga cgttgaccaa agttcttgct ctcgtcaaag aggttcctcg
     2041 taatgatatc aatatttatc cggaatacat tctgccaggc atagcccact tagcccaaga
     2101 tgatgctact atcgttagac tagcctatgc tgaaaacata gctctgctgg cagaaacagc
     2161 tctgagattc ctggaattag tacagttaaa aaatcttaat atggaaaatg accccaataa
     2221 tgaagaaata gatgaggtta cacatccaaa tggaaattat gacacagagc tccaagcctt
     2281 acatgaaatg gtccagcaga aagttgttac tttgctaagt gaccctgaaa atattgtaaa
     2341 acaaaccttg atggaaaatg gaataacacg gctgtgtgta ttctttggac gtcagaaagc
     2401 caacgatgtt ttgttgtccc acatgattac tttcctaaat gataagaatg attggcatct
     2461 acgtggagca ttttttgata gtatagttgg tgttgctgcc tatgttggct ggcaaagctc
     2521 ctcaattctc aagcctctgc tgcaacaagg tcttagtgat gctgaggaat ttgtcattgt
     2581 gaaagctctt tatgccctta cttgtatgtg ccagttagga ctgctacaaa aaccccatgt
     2641 ttacgaattt gccagtgata ttgccccctt cctgtgtcat cccaatttat ggatacgtta
     2701 tggtgccgtg ggatttatca cagtggtagc tcgtcaaata agtacagctg atgtctactg
     2761 taaactgatg ccttatcttg acccatatat tacccaacca ataatacaga ttgaaagaaa
     2821 acttgttctg ctcagtgttt taaaggaacc agtaagtcgt tctatatttg attatgcttt
     2881 gaggtctaaa gatattacta gcttgttcag acatcttcac atgcgtcaga agaaacgaaa
     2941 tggttctctt cccgactgcc ctccgccaga ggatcctgcc atagcacagc ttctgaagaa
     3001 gttgctctca cagggaatga cagaggaaga ggaagacaaa cttctggcac tgaaagactt
     3061 catgatgaaa tctaataaag caaaggccaa tatagtggac cagagccatc ttcatgatag
     3121 tagtcagaaa ggtgtaattg acttggcagc tttaggcata actgggagac aagttgatct
     3181 tgttaaaacc aaacaagaac cagatgacaa acgggccaga aaacatgtaa aacaagactc
     3241 aaatgtaaat gaagaatgga aaagcatgtt tgggtcactg gacccaccaa acatgccaca
     3301 ggccctacct aaagggagtg atcaggaggt gattcagact gggaaacctc ctcgttccga
     3361 gtcctctgct ggcatttgtg tccctttgtc aacttcttca caggttccag aagtgacaac
     3421 tgtccaaaat aaaaaaccag taataccggt tttaagtagt acaatcttac catccaccta
     3481 tcagattcga attacaactt gtaaaactga acttcagcaa ctcatccagc aaaagcggga
     3541 gcagtgcaat gctgagagaa tagctaagca gatgatggaa aatgctgaat gggagagtaa
     3601 accaccacca cctggatggc gtcctaaagg gctgttagtt gcccatcttc atgagcataa
     3661 atctgctgtg aatcgaatta gagtctctga tgaacactca ctttttgcaa catgttcaaa
     3721 tgatggcaca gtgaaaatct ggaacagtca aaagatggag gggaagacca ccactaccag
     3781 atctattctt acatacagcc gaattggagg acgagtcaag acgctcacat tctgccaagg
     3841 ctcccactat ttagccatag catctgataa tggtgctgtc cagcttcttg gaattgaggc
     3901 ttctaagctg cccaagtctc ctaaaatcca tcctctacaa agcagaattc tagatcagaa
     3961 ggaggacggt tgtgttgtgg atatgcatca cttcaactct ggagcacagt ctgttcttgc
     4021 ctatgccact gtgaatggct ctctggttgg ctgggacctt aggtcttcaa gcaatgcgtg
     4081 gactttaaag catgatttaa agtcgggcct catcacttcc tttgctgtgg acatccacca
     4141 atgctggctc tgcattggta caagcagtgg taccatggct tgttgggaca tgaggttcca
     4201 gttgccaatt tcaagtcact gtcatccttc cagggctcga atcagacgcc tctcaatgca
     4261 ccctctgtat cagtcctggg tgattgcagc tgttcagggc aacaacgaag tgtccatgtg
     4321 ggacatggag actggtgaca gaagatttac tctctgggcc agcagtgcac caccactttc
     4381 tgaattacag ccttctcctc atagcgtcca tggtatctac tgtagtcctg cagatggaaa
     4441 tcctatccta ctaacagctg gctcagatat gaaaataagg ttttgggact tggcttaccc
     4501 agaaaggtcc tatgttgttg caggaagtac tagttcccca tctgtgtcct actacaggaa
     4561 aataattgaa ggcactgaag ttgtccagga aattcagaat aagcagaaag taggaccaag
     4621 tgatgacacc cctcgaaggg gcccagagtc cctgcccgtg ggacatcatg acatcatcac
     4681 tgatgtcgcc acattccaga ccacacaggg cttcatcgta actgcttcta gagatgggat
     4741 tgtgaaggtg tggaaactcg aggactacaa agaccatgac ggtgattata aagatcatga
     4801 catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga caatcaacct
     4861 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     4921 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     4981 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     5041 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     5101 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     5161 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     5221 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt
     5281 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     5341 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     5401 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg gggaggctaa
     5461 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     5521 agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     5581 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     5641 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     5701 gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta tccccacgcg
     5761 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     5821 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     5881 gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct
     5941 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     6001 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     6061 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     6121 attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     6181 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     6241 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     6301 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     6361 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     6421 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     6481 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     6541 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     6601 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     6661 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     6721 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     6781 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     6841 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     6901 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     6961 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     7021 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     7081 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     7141 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     7201 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     7261 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     7321 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     7381 tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca
     7441 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     7501 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     7561 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     7621 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     7681 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     7741 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     7801 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     7861 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     7921 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     7981 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     8041 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     8101 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     8161 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     8221 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     8281 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc
     8341 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     8401 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     8461 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     8521 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     8581 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     8641 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     8701 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     8761 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     8821 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     8881 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     8941 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     9001 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     9061 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     9121 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     9181 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     9241 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     9301 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     9361 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     9421 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     9481 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     9541 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     9601 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     9661 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     9721 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     9781 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     9841 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     9901 ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat gccgcatagt
     9961 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
    10021 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagg
//
