LOCUS       V020174                 8111 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020174
VERSION     V020174
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             941..3601
                     /label="PIK3C3(NM_002647)"
                     /note="PIK3C3(NM_002647)"
                     /gene="PIK3C3"
     CDS             3608..3673
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3711..3935
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3981..4409
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4423..4752
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4819..5613
                     /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    5787..5908
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5957..5973)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5981..5997
                     /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(6005..6035)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6050..6071
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6359..6944)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7115..7975)
                     /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(7976..8080)
                     /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 atccgccacc atgggggaag cagagaagtt
      961 tcactacatc tatagttgtg acctggatat caacgtccag cttaagatag gaagcttgga
     1021 agggaagaga gaacaaaaga gttataaagc tgtcctggaa gacccaatgt tgaagttctc
     1081 aggactatat caagagacat gctctgatct ttatgttact tgtcaagttt ttgcagaagg
     1141 gaagcctttg gccttgccag tgagaacatc ctacaaagca tttagtacaa gatggaactg
     1201 gaatgaatgg ctgaaactac cagtaaaata ccctgacctg cccaggaatg cccaagtggc
     1261 cctcaccata tgggatgtgt atggtcccgg aaaagcagtg cctgtaggag gaacaacggt
     1321 ttcgctcttt ggaaaatacg gcatgtttcg ccaagggatg catgacttga aagtctggcc
     1381 taatgtagaa gcagatggat cagaacccac aaaaactcct ggcagaacaa gtagcactct
     1441 ctcagaagat cagatgagcc gtcttgccaa gctcaccaaa gctcatcgac aaggacacat
     1501 ggtgaaagta gattggctgg atagattgac atttagagaa atagaaatga taaatgagag
     1561 tgaaaaacga agttctaatt tcatgtacct gatggttgaa tttcgatgtg tcaagtgtga
     1621 tgataaggaa tatggtattg tttattatga aaaggacggt gatgaatcat ctccaatttt
     1681 aacaagtttt gaattagtga aagttcctga cccccagatg tctatggaga atttagttga
     1741 gagcaaacac cacaagcttg cccggagttt aagaagtgga ccttctgacc acgatctgaa
     1801 acccaatgct gccacgagag atcagttaaa tattattgtg agttatccac caaccaagca
     1861 acttacatat gaagaacaag atcttgtttg gaagtttaga tattatctta cgaatcaaga
     1921 aaaagccttg acaaaattct tgaaatgtgt taattgggat ctacctcaag aggccaaaca
     1981 ggccttggaa cttctgggaa aatggaagcc gatggatgta gaggactcct tggagctgtt
     2041 atcctctcat tacaccaacc caactgtgag gcgttatgct gttgcccggt tgcgacaggc
     2101 cgatgatgag gatttgttga tgtacctatt acaattggtc caggctctca aatatgaaaa
     2161 ttttgatgat ataaagaatg gattggaacc taccaagaag gatagtcaga gttcagtgtc
     2221 agaaaatgtg tcaaattctg gaataaattc tgcagaaata gatagctccc aaattataac
     2281 cagccccctt ccttcagtct cttcacctcc tcctgcatca aaaacaaaag aagttccaga
     2341 tggcgaaaat ctggaacaag atctctgtac cttcttgata tcgagagcct gcaaaaactc
     2401 aacactggct aattatttat actggtatgt gatagtggaa tgtgaagatc aagatactca
     2461 gcagagagat ccaaagaccc atgagatgta cttgaacgta atgagaagat tcagccaagc
     2521 attgttgaag ggtgataagt ctgtcagagt tatgcgttct ttgctggctg cacaacagac
     2581 atttgtagat cggttggtgc atctaatgaa ggcagtacaa cgcgaaagtg gaaatcgtaa
     2641 gaaaaagaat gagagactac aggcattgct tggagataat gaaaagatga atttgtcaga
     2701 tgtggaactt atcccgttgc ctttagaacc ccaagtgaaa attagaggaa taattccgga
     2761 aacagctaca ctgtttaaaa gtgcccttat gcctgcacag ttgtttttta agacggaaga
     2821 tggaggcaaa tatccagtta tatttaagca tggagatgat ttacgtcaag atcaacttat
     2881 tcttcaaatc atttcactca tggacaagct gttacggaaa gaaaatctgg acttgaaatt
     2941 gacaccttat aaggtgttag ccaccagtac aaaacatggc ttcatgcagt ttatccagtc
     3001 agttcctgtg gctgaagttc ttgatacaga gggaagcatt cagaactttt ttagaaaata
     3061 tgcaccaagt gagaatgggc caaatgggat tagtgctgag gtcatggaca cttacgttaa
     3121 aagctgtgct ggatattgcg tgatcaccta tatacttgga gttggagaca ggcacctgga
     3181 taaccttttg ctaacaaaaa caggcaaact cttccacata gactttggat atattttggg
     3241 tcgggatcca aagcctcttc ctccaccaat gaagctgaat aaagaaatgg tagaaggaat
     3301 ggggggcaca cagagtgagc agtaccaaga gttccgtaaa cagtgttaca cggctttcct
     3361 ccacctgcga aggtattcta atctgatttt gaacttgttt tccttgatgg ttgatgcaaa
     3421 cattccagat attgcacttg aaccagataa aactgtgaaa aaggttcagg ataaattccg
     3481 cttagacctg tcggatgaag aggctgtgca ttacatgcag agtctgattg atgagagtgt
     3541 ccatgctctt tttgctgcag tggtggaaca gattcacaag tttgcccagt actggagaaa
     3601 actcgaggac tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga
     3661 tgacgatgac aagtagtgag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     3721 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     3781 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     3841 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     3901 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     3961 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4021 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     4081 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4141 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     4201 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4261 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     4321 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     4381 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     4441 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4501 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4561 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     4621 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     4681 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4741 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     4801 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     4861 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     4921 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     4981 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     5041 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     5101 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     5161 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     5221 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     5281 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     5341 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     5401 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     5461 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     5521 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     5581 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     5641 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     5701 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     5761 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     5821 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     5881 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     5941 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     6001 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     6061 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     6121 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     6181 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     6241 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     6301 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     6361 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     6421 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     6481 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     6541 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6601 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6661 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6721 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6781 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6841 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt
     6901 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6961 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     7021 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     7081 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     7141 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     7201 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     7261 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     7321 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     7381 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     7441 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     7501 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     7561 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     7621 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     7681 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     7741 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     7801 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     7861 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7921 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7981 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     8041 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     8101 cacctgacgt c
//
