LOCUS       V019146                10394 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019146
VERSION     V019146
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..5074
                     /label="Grin2a(NM_012573)"
                     /note="Grin2a(NM_012573)"
                     /gene="Grin2a"
     CDS             5081..5146
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    5170..5758
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5824..5872
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      6074..6502
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6516..6845
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6912..7706
                     /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    7882..8003
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(8052..8068)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    8076..8092
                     /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(8100..8130)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    8145..8166
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8454..9042)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9213..10073)
                     /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(10074..10178)
                     /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 ccatgggcag attgggctac tggaccttgc tggtattgcc
      721 ggcccttctg gtctggcgcg atccggcgca gaacgcggcg gcggagaagg gtcctccagc
      781 gctgaacatt gcggtgctgc tgggtcacag ccacgacgtg acagaacgcg aacttcgaaa
      841 tctgtggggc ccagagcagg caaccggctt gcccctggat gtgaacgtgg tggcgttatt
      901 gatgaaccgc actgacccta agagcctcat cacgcatgtg tgcgacctca tgtccggggc
      961 gcgcatccac ggcttggtgt ttggagatga cacggaccag gaggctgtgg cccagatgct
     1021 ggattttatc tcctcacaga cttttatccc catcttgggc attcatgggg gtgcatctat
     1081 gatcatggct gacaaggatc cgacatccac gttcttccag tttggagcct ccatccagca
     1141 gcaagccaca gttatgctga agatcatgca ggactacgac tggcacgtct tctccctggt
     1201 caccaccatc ttccctggct accgagactt catcagcttt atcaagacaa cagtggacaa
     1261 cagctttgtg ggctgggata tgcagaacgt gatcacactg gacacctcct tcgaggacgc
     1321 caagacgcag gtccagctga agaagatcca ttcttctgtc atcctgctct actgctccaa
     1381 agacgaggct gtcctcatcc tgagcgaggc tcgctccctc ggcctcactg gctatgattt
     1441 cttctggatt gtccccagtt tggtgtctgg gaacacagag ctcatcccca aagagtttcc
     1501 atcaggtctc atttcagtct cttatgacga ctgggactac agcctggagg caagagtgag
     1561 agacggtctt gggatcttaa ccactgccgc atcctccatg ttggagaagt tctcctacat
     1621 tcctgaggcc aaggccagct gctatgggca ggcagagaag ccagagaccc cgctacacac
     1681 cctgcaccaa ttcatggtca atgtgacttg ggatggcaag gacttgtcct tcactgagga
     1741 aggttaccag gtgcacccca ggcttgtggt gatcgtgctg aacaaggacc gggagtggga
     1801 aaaggtgggc aagtgggaga accagacgct gagcctgagg cacgctgtgt ggccaaggta
     1861 caagtccttt tctgactgcg agccagatga caaccacctc agcattgtca ccttggagga
     1921 agcccccttc gtcatcgtag aggacataga ccccctgact gagacctgtg tgaggaacac
     1981 ggtgccctgt cggaagttcg tcaagatcaa caattcaacc aacgaaggga tgaatgtgaa
     2041 gaaatgctgc aaggggttct gcatcgacat cctcaagaag ctgtccagaa ctgtgaagtt
     2101 cacctatgac ctctacctgg tgaccaatgg gaagcatggg aaaaaggtta acaatgtgtg
     2161 gaatggaatg ataggtgaag tggtctatca acgagcagtc atggctgtgg gctcactcac
     2221 catcaatgag gagcgttcgg aagtggtgga cttctcggtg cccttcgtgg agacaggaat
     2281 cagcgtcatg gtctccagga gtaatggcac tgtctcccct tctgctttcc tcgaaccctt
     2341 cagtgcctcc gtctgggtga tgatgtttgt gatgctgctc atcgtctcag ccattgctgt
     2401 cttcgttttt gaatacttca gtcctgttgg atacaacaga aacttagcca aagggaaagc
     2461 tccccacggg ccttctttta ctattggaaa agctatatgg ctcctctggg gcctggtctt
     2521 caacaattct gtgcctgtcc agaatcctaa aggcacaacc agcaagatca tggtgtcagt
     2581 gtgggccttc tttgctgtca tcttcctggc cagttacaca gccaacctgg ctgccttcat
     2641 gatccaggag gagtttgtgg accaagtgac tggcctcagt gacaagaagt tccagagacc
     2701 tcatgactat tctccacctt tccgatttgg gacggtaccc aatggaagta cagagaggaa
     2761 tattcgtaac aactacccgt atatgcacca gtacatgacc agattcaacc agaggggagt
     2821 ggaggatgcc ttggtcagct tgaaaaccgg gaagttggac gctttcatct atgacgcagc
     2881 cgtcttgaac tacaaggccg ggagggatga aggctgtaaa ctggtgacca ttgggagcgg
     2941 gtacatcttt gctaccacag gctatggaat tgcgctgcag aagggctcac cctggaagag
     3001 gcagattgac ctcgctctgc tccagtttgt tggtgatggt gagatggagg agctggagac
     3061 actgtggctt acgggcatct gccacaacga gaagaatgag gtgatgagta gccagctgga
     3121 catcgataac atggcgggcg tgttctacat gctggctgca gccatggccc tcagcctcat
     3181 caccttcatc tgggagcacc tcttctactg gaagctgcgc ttctgcttca caggcgtgtg
     3241 ctctgaccgg cccgggctgc tcttctccat cagcaggggc atctatagtt gcatccatgg
     3301 ggtacacatt gaagaaaaga agaaatctcc agatttcaat ctgactggat cacagagcaa
     3361 catgctaaag cttcttcggt cagctaaaaa catctccaat atgtccaaca tgaactcctc
     3421 aagaatggac tcacctaaaa gagctactga cttcattcaa agagggtcac ttattgtgga
     3481 catggtttca gacaagggaa atttgatata ttcagacaac agatcctttc aagggaagga
     3541 cagtatattt ggagacaaca tgaatgaact ccaaacattt gtggccaaca ggcacaagga
     3601 taatctcagt aactatgtgt ttcaaggaca gcatcctctc actctcaatg agtccaaccc
     3661 taacacagta gaggtggctg tcagcactga atccaaaggg aactcccgac cccggcagct
     3721 ttggaagaaa tccatggagt ctctacgcca ggattctcta aaccagaacc cagtctccca
     3781 gagggatgag aagactgcag agaatcggac ccactcgcta aagagtccta ggtatcttcc
     3841 agaagaggta gcccactctg acatttcaga aacctcaagc cgggccacat gccacaggga
     3901 gccagataac aataagaacc acaagaccaa ggataacttc aaacggtcaa tggcctctaa
     3961 gtatcccaag gactgtagcg atgttgaccg cacctacatg aaaaccaaag caagttctcc
     4021 cagggataag atctatacca ttgatggtga gaaggagccc agcttccact tagatcctcc
     4081 tcagtttgtt gagaatataa ccctgcctga gaatgtgggc ttcccagata cctaccaaga
     4141 tcacaatgag aacttccgca agggggactc cacactgccc atgaacagga acccattaca
     4201 taatgaagac gggcttccca acaatgacca atataaactc tatgccaagc actttacctt
     4261 gaaagacaag ggttccccac acagtgaggg cagtgatcga taccggcaga actccacaca
     4321 ttgcagaagc tgcctttcga atctgcccac ctactcaggc cactttacca tgaggtctcc
     4381 tttcaagtgt gatgcctgtc tgcggatggg gaatctctat gacattgatg aagaccagat
     4441 gcttcaggag acaggtaacc cagctactcg ggaggaggtc taccagcagg actggtcaca
     4501 gaacaacgcc ctccagttcc agaagaacaa gctaaggatt aaccgacagc actcctatga
     4561 taacattctg gacaaaccca gagagataga ccttagcagg ccctcccgga gcataagcct
     4621 caaggacagg gaacggctac tggagggcaa cttgtatggg agcctgttca gtgtcccctc
     4681 aagcaaactc ttggggaaca aaagctccct tttcccccaa ggtctggagg acagcaagag
     4741 gagcaagtct ctcttgccag accacgcctc cgataatcct ttcctccaca cgtatgggga
     4801 tgaccaacgc ttagttatcg ggagatgtcc ctcggaccct tacaaacact cattgccatc
     4861 acaggcggta aatgacagct atcttcggtc atccttgagg tcaacagcat catattgctc
     4921 cagggacagt cggggccaca gtgatgtgta tatttcagag catgttatgc cttatgctgc
     4981 aaataagaat accatgtact ctacccccag ggttttaaat tcctgcagca atagacgagt
     5041 gtacaagaaa atgcctagta tcgaatctga tgttctcgag gactacaaag accatgacgg
     5101 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccga
     5161 tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     5221 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     5281 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     5341 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     5401 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     5461 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     5521 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
     5581 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     5641 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     5701 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     5761 ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     5821 tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc
     5881 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     5941 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     6001 ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct
     6061 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     6121 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     6181 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg catcccttta
     6241 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     6301 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     6361 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     6421 tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa tgagctgatt
     6481 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     6541 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     6601 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     6661 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     6721 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     6781 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     6841 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     6901 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     6961 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     7021 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     7081 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     7141 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     7201 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     7261 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     7321 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     7381 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     7441 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     7501 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     7561 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     7621 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     7681 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa
     7741 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     7801 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     7861 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     7921 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     7981 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     8041 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     8101 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     8161 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     8221 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     8281 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     8341 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     8401 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     8461 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     8521 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     8581 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     8641 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     8701 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     8761 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     8821 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     8881 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     8941 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     9001 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     9061 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     9121 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     9181 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     9241 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     9301 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     9361 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     9421 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     9481 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     9541 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     9601 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     9661 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     9721 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     9781 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     9841 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     9901 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     9961 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
    10021 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
    10081 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
    10141 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
    10201 cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc agtacaatct
    10261 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
    10321 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
    10381 agaatctgct tagg
//
