LOCUS       V028527                 8199 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028527
VERSION     V028527
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..2879
                     /label="PJA2(NM_014819)"
                     /note="PJA2(NM_014819)"
                     /gene="PJA2"
     CDS             2886..2951
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2975..3563
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3629..3677
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3879..4307
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4321..4650
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4717..5511
                     /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    5687..5808
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5857..5873)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5881..5897
                     /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(5905..5935)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5950..5971
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6259..6847)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7018..7878)
                     /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(7879..7983)
                     /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 ccaccatgtc acagtacact gaaaaggagc
      781 cagcagcaat ggaccaagaa tctggtaagg ctgtctggcc caaaccagca ggagggtatc
      841 agacaattac aggcaggaga tatggaagaa gacatgctta tgtcagtttt aaaccatgta
      901 tgaccagaca tgaaagaagc ttaggtcggg ctggtgatga ctatgaagtg ttggaactag
      961 atgatgttcc aaaggaaaat tcctcaggtt ccagtccttt ggatcaagtt gattcttctt
     1021 tacccagtga acctatattt gaaaaaagtg aaacagaaat tcccacttgt ggttcagcat
     1081 tgaatcaaac cactgagagc agtcaatcct ttgttgcagt acatcacagt gaggaaggca
     1141 gggatacctt aggaagcagt acaaatcttc ataatcactc tgagggagag tatattccag
     1201 gagcttgtag tgcttcaagt gtccaaaatg gaattgcatt ggttcataca gactcttatg
     1261 atccagatgg caaacatgga gaagataatg accatcttca actttctgca gaagtcgtgg
     1321 aaggtagtag ataccaggaa tcattaggca atacagtatt tgagttggaa aacagagagg
     1381 cagaggcata cactggtctt tcaccaccag ttccctcatt taactgtgaa gtaagagatg
     1441 agtttgaaga gttagattct gtaccattag tgaaaagttc tgctggtgat actgagtttg
     1501 tccatcagaa tagccaggaa attcagaggt cttctcaaga tgaaatggtt agtacgaaac
     1561 aacaaaataa tactagccag gaaagacaga cagaacattc acctgaagat gcagcctgtg
     1621 gtccagggca tatttgtagt gaacaaaata ccaatgatag ggaaaagaac catggaagtt
     1681 ctcctgaaca ggtagtgagg ccaaaagtta gaaaactgat aagttcaagc caggtggacc
     1741 aagaaacagg ttttaatagg catgaggcga aacaaagaag tgttcaaaga tggagagagg
     1801 ctttggaagt tgaggaaagt ggctcagatg acctcttaat aaaatgtgaa gaatatgatg
     1861 gagagcatga ctgtatgttc ttggatccac catactcaag agttattaca caaagggaaa
     1921 cagaaaataa ccaaatgaca tcagaaagtg gagccacagc aggaaggcaa gaggtggata
     1981 acaccttttg gaatggctgt ggagattatt accaactcta tgacaaagat gaagatagtt
     2041 ctgaatgcag tgatggggaa tggtctgctt ctttgcctca tcgattttct ggtacagaaa
     2101 aagatcaatc ctcaagtgat gaaagctggg agactctgcc aggaaaagat gagaatgaac
     2161 ctgagctaca aagtgatagc agtggccctg aagaagaaaa ccaagaatta tctcttcagg
     2221 aaggggaaca gacatccttg gaagagggag aaattccttg gttacagtac aatgaagtca
     2281 atgaaagcag cagtgatgag ggaaatgaac ctgccaatga atttgcacag ccagctttca
     2341 tgttggatgg taacaataac ctggaggatg actccagtgt gagtgaagac ttagatgtgg
     2401 attggagcct atttgatggc tttgcagatg gactaggagt tgctgaagct atttcatatg
     2461 tggatcctca gttccttacc tacatggcac tagaagaacg cttagcccag gctatggaga
     2521 ctgctctggc ccatttagag tctcttgcag tggatgttga ggtggccaat ccaccagcta
     2581 gtaaggaaag cattgatggt cttccagaga cccttgttct tgaagatcac actgctattg
     2641 gtcaggaaca atgctgtcca atctgttgca gtgagtatat taaggatgat atagcaacag
     2701 agttgccctg tcaccatttc tttcacaaac cttgtgtctc aatttggcta caaaagtcgg
     2761 gaacatgccc tgtgtgccgc cgtcatttcc cacctgcggt tattgaagca tctgcagctc
     2821 cttcctctga gcctgatcct gatgccccac cttcaaatga cagtattgca gaagcacccg
     2881 gatccgacta caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg
     2941 acgatgacaa gtagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg
     3001 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     3061 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     3121 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     3181 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     3241 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     3301 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     3361 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     3421 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     3481 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     3541 ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag
     3601 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg
     3661 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     3721 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     3781 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     3841 agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     3901 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3961 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4021 agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4081 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4141 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4201 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc
     4261 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     4321 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4381 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     4441 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4501 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4561 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     4621 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     4681 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4741 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4801 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4861 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     4921 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4981 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     5041 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     5101 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     5161 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     5221 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     5281 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5341 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5401 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     5461 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5521 tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5581 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5641 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     5701 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5761 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     5821 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5881 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5941 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     6001 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     6061 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6121 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6181 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6241 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6301 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6361 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6421 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     6481 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6541 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6601 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6661 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     6721 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6781 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6841 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6901 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6961 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     7021 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7081 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7141 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7201 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7261 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7321 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7381 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7441 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7501 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7561 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7621 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7681 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7741 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7801 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7861 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7921 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7981 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     8041 ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     8101 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     8161 caaggcttga ccgacaattg catgaagaat ctgcttagg
//
