LOCUS       V022280                 8705 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022280
VERSION     V022280
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..3352
                     /label="PRKD3(NM_005813)"
                     /note="PRKD3(NM_005813)"
                     /gene="PRKD3"
     CDS             3392..3457
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3481..4069
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4135..4183
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4385..4813
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4827..5156
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5223..6017
                     /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    6193..6314
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6363..6379)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6387..6403
                     /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(6411..6441)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6456..6477
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6765..7353)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7524..8384)
                     /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(8385..8489)
                     /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 ccatgtctgc aaataattcc cctccatcag cccagaagtc
      721 tgtattaccc acagctattc ctgctgtgct tccagctgct tctccgtgtt caagtcctaa
      781 gacgggactc tctgcccgac tctctaatgg aagcttcagt gcaccatcac tcaccaactc
      841 cagaggctca gtgcatacag tttcatttct actgcaaatt ggcctcacac gggagagtgt
      901 taccattgaa gcccaggaac tgtctttatc tgctgtcaag gatcttgtgt gctccatagt
      961 ttatcaaaag tttccagagt gtggattctt tggcatgtat gacaaaattc ttctctttcg
     1021 ccatgacatg aactcagaaa acattttgca gctgattacc tcagcagatg aaatacatga
     1081 aggagaccta gtggaagtgg ttctttcagc tttagccaca gtagaagact tccagattcg
     1141 tccacatact ctctatgtac attcttacaa agctcctact ttctgtgatt actgtggtga
     1201 gatgctctgg ggattggtac gtcaaggact gaaatgtgaa ggctgtggat taaattacca
     1261 taaacgatgt gccttcaaga ttccaaataa ctgtagtgga gtaagaaaga gacgtctgtc
     1321 aaatgtatct ttaccaggac ccggcctctc agttccaaga cccctacagc ctgaatatgt
     1381 agcccttccc agtgaagagt cacatgtcca ccaggaacca agtaagagaa ttccttcttg
     1441 gagtggtcgc ccaatctgga tggaaaagat ggtaatgtgc agagtgaaag ttccacacac
     1501 atttgctgtt cactcttaca cccgtcccac gatatgtcag tactgcaagc ggttactgaa
     1561 aggcctcttt cgccaaggaa tgcagtgtaa agattgcaaa ttcaactgcc ataaacgctg
     1621 tgcatcaaaa gtaccaagag actgccttgg agaggttact ttcaatggag aaccttccag
     1681 tctgggaaca gatacagata taccaatgga tattgacaat aatgacataa atagtgatag
     1741 tagtcggggt ttggatgaca cagaagagcc atcaccccca gaagataaga tgttcttctt
     1801 ggatccatct gatctcgatg tggaaagaga tgaagaagcc gttaaaacaa tcagtccatc
     1861 aacaagcaat aatattccgc taatgagggt tgtacaatcc atcaagcaca caaagaggaa
     1921 gagcagcaca atggtgaagg aagggtggat ggtccattac accagcaggg ataacctgag
     1981 aaagaggcat tattggagac ttgacagcaa atgtctaaca ttatttcaga atgaatctgg
     2041 atcaaagtat tataaggaaa ttccactttc agaaattctc cgcatatctt caccacgaga
     2101 tttcacaaac atttcacaag gcagcaatcc acactgtttt gaaatcatta ctgatactat
     2161 ggtatacttc gttggtgaga acaatgggga cagctctcat aatcctgttc ttgctgccac
     2221 tggagttgga cttgatgtag cacagagctg ggaaaaagca attcgccaag ccctcatgcc
     2281 tgttactcct caagcaagtg tttgcacttc tccagggcaa gggaaagatc acaaagattt
     2341 gtctacaagt atctctgtat ctaattgtca gattcaggag aatgtggata tcagtactgt
     2401 ttaccagatc tttgcagatg aggtgcttgg ttcaggccag tttggcatcg tttatggagg
     2461 aaaacataga aagactggga gggatgtggc tattaaagta attgataaga tgagattccc
     2521 cacaaaacaa gaaagtcaac tccgtaatga agtggctatt ttacagaatt tgcaccatcc
     2581 tgggattgta aacctggaat gtatgtttga aaccccagaa cgagtctttg tagtaatgga
     2641 aaagctgcat ggagatatgt tggaaatgat tctatccagt gagaaaagtc ggcttccaga
     2701 acgaattact aaattcatgg tcacacagat acttgttgct ttgaggaatc tgcattttaa
     2761 gaatattgtg cactgtgatt taaagccaga aaatgtgctg cttgcatcag cagagccatt
     2821 tcctcaggtg aagctgtgtg actttggatt tgcacgcatc attggtgaaa agtcattcag
     2881 gagatctgtg gtaggaactc cagcatactt agcccctgaa gttctccgga gcaaaggtta
     2941 caaccgttcc ctagatatgt ggtcagtggg agttatcatc tatgtgagcc tcagtggcac
     3001 atttcctttt aatgaggatg aagatataaa tgaccaaatc caaaatgctg catttatgta
     3061 cccaccaaat ccatggagag aaatttctgg tgaagcaatt gatctgataa acaatctgct
     3121 tcaagtgaag atgagaaaac gttacagtgt tgacaaatct cttagtcatc cctggctaca
     3181 ggactatcag acttggcttg accttagaga atttgaaact cgcattggag aacgttacat
     3241 tacacatgaa agtgatgatg ctcgctggga aatacatgca tacacacata accttgtata
     3301 cccaaagcac ttcattatgg ctcctaatcc agatgatatg gaagaagatc ctgaattctg
     3361 cagatatcca gcacagtggc ggccgctcga ggactacaaa gaccatgacg gtgattataa
     3421 agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg atatctcgac
     3481 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     3541 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     3601 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     3661 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     3721 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     3781 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     3841 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc
     3901 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     3961 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     4021 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaaacggg
     4081 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     4141 taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg
     4201 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     4261 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     4321 aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc tagggggtat
     4381 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4441 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     4501 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga
     4561 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     4621 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4681 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     4741 ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4801 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4861 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     4921 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4981 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     5041 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     5101 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     5161 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     5221 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     5281 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     5341 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     5401 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     5461 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     5521 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     5581 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     5641 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5701 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     5761 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5821 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     5881 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5941 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     6001 ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc
     6061 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     6121 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     6181 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     6241 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     6301 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     6361 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     6421 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     6481 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     6541 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     6601 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     6661 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     6721 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     6781 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     6841 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     6901 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa
     6961 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     7021 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     7081 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     7141 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     7201 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     7261 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     7321 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     7381 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     7441 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     7501 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     7561 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     7621 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     7681 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     7741 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     7801 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     7861 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     7921 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     7981 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     8041 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     8101 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     8161 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     8221 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     8281 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     8341 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     8401 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     8461 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     8521 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
     8581 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
     8641 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
     8701 ttagg
//
