LOCUS       V020343                 9701 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020343
VERSION     V020343
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..4381
                     /label="GRIN2C(NM_000835)"
                     /note="GRIN2C(NM_000835)"
                     /gene="GRIN2C"
     CDS             4388..4453
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4477..5065
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5131..5179
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5381..5809
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5823..6152
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6219..7013
                     /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    7189..7310
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7359..7375)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7383..7399
                     /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(7407..7437)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7452..7473
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7761..8349)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8520..9380)
                     /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(9381..9485)
                     /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 ccatgggtgg ggccctgggg ccggccctgt tgctcacctc
      721 gctcttcggt gcctgggcag ggctgggtcc ggggcagggc gagcagggca tgacggtggc
      781 cgtggtgttt agcagctcag ggccgcccca ggcccagttc cgtgcccgcc tcacccccca
      841 gagcttcctg gacctacccc tggagatcca gccgctcaca gttggggtca acaccaccaa
      901 ccccagcagc ctcctcaccc agatctgcgg cctcctgggt gctgcccacg tccacggcat
      961 tgtctttgag gacaacgtgg acaccgaggc ggtggcccag atccttgact tcatctcctc
     1021 ccagacccat gtgcccatcc tcagcatcag cggaggctct gctgtggtcc tcacccccaa
     1081 ggagccgggc tccgccttcc tgcagctggg cgtgtccctg gagcagcagc tgcaggtgct
     1141 gttcaaggtg ctggaagagt acgactggag cgccttcgcc gtcatcacca gcctgcaccc
     1201 gggccacgcg ctcttcctgg agggcgtgcg cgccgtcgcc gacgccagcc acgtgagttg
     1261 gcggctgctg gacgtggtca cgctggagct gggcccggga gggccgcgcg cgcgcacgca
     1321 gcgcctgctg cgccagctcg acgcgcccgt gtttgtggcc tactgctcgc gcgaggaggc
     1381 cgaggtgctc ttcgccgagg cggcgcaggc cggtctggtg gggcccggcc acgtgtggct
     1441 ggtgcccaac ctggcgctgg gcagcaccga tgcgcccccc gccaccttcc ccgtgggcct
     1501 catcagcgtc gtcaccgaga gctggcgcct cagcctgcgc cagaaggtgc gcgacggcgt
     1561 ggccattctg gccctgggcg cccacagcta ctggcgccag catggaaccc tgccagcccc
     1621 ggccggggac tgccgtgttc accctgggcc cgtcagccct gcccgggagg ccttctacag
     1681 gcacctactg aatgtcacct gggagggccg agacttctcc ttcagccctg gtgggtacct
     1741 ggtccagccc accatggtgg tgatcgccct caaccggcac cgcctctggg agatggtggg
     1801 gcgctgggag catggcgtcc tatacatgaa gtaccccgtg tggcctcgct acagtgcctc
     1861 tctgcagcct gtggtggaca gtcggcacct gacggtggcc acgctggaag agcggccctt
     1921 tgtcatcgtg gagagccctg accctggcac aggaggctgt gtccccaaca ccgtgccctg
     1981 ccgcaggcag agcaaccaca ccttcagcag cggggacgtg gccccctaca ccaagctctg
     2041 ctgtaaggga ttctgcatcg acatcctcaa gaagctggcc agagtggtca aattctccta
     2101 cgacctgtac ctggtgacca acggcaagca tggcaagcgg gtgcgcggcg tatggaacgg
     2161 catgattggg gaggtgtact acaagcgggc agacatggcc atcggctccc tcaccatcaa
     2221 tgaggaacgc tccgagatcg tagacttctc tgtacccttt gtggagacgg gcatcagtgt
     2281 gatggtggct cgcagcaatg gcaccgtctc cccctcggcc ttcttggagc catatagccc
     2341 tgcagtgtgg gtgatgatgt ttgtcatgtg cctcactgtg gtggccatca ccgtcttcat
     2401 gttcgagtac ttcagccctg tcagctacaa ccagaacctc accagaggca agaagtccgg
     2461 gggcccagct ttcactatcg gcaagtccgt gtggctgctg tgggcgctgg tcttcaacaa
     2521 ctcagtgccc atcgagaacc cgcggggcac caccagcaag atcatggttc tggtctgggc
     2581 cttctttgct gtcatcttcc tcgccagcta cacggccaac ctggccgcct tcatgatcca
     2641 agagcaatac atcgacactg tgtcgggcct cagtgacaag aagtttcagc ggcctcaaga
     2701 tcagtaccca cctttccgct tcggcacggt gcccaacggc agcacggagc ggaacatccg
     2761 cagtaactac cgtgacatgc acacccacat ggtcaagttc aaccagcgct cggtggagga
     2821 cgcgctcacc agcctcaaga tggggaagct ggatgccttc atctatgatg ctgctgtcct
     2881 caactacatg gcaggcaagg acgagggctg caagctggtc accattgggt ctggcaaggt
     2941 ctttgctacc actggctacg gcatcgccat gcagaaggac tcccactgga agcgggccat
     3001 agacctggcg ctcttgcagt tcctggggga cggagagaca cagaaactgg agacagtgtg
     3061 gctctcaggg atctgccaga atgagaagaa cgaggtgatg agcagcaagc tggacatcga
     3121 caacatggca ggcgtcttct acatgctgct ggtggccatg gggctggccc tgctggtctt
     3181 cgcctgggag cacctggtct actggaagct gcgccactcg gtgcccaact catcccagct
     3241 ggacttcctg ctggctttca gcaggggcat ctacagctgc ttcagcgggg tgcagagcct
     3301 cgccagccca ccgcggcagg ccagcccgga cctcacggcc agctcggccc aggccagcgt
     3361 gctcaagatg ctgcaggcag cccgcgacat ggtgaccacg gcgggcgtaa gcagctccct
     3421 ggaccgcgcc actcgcacca tcgagaattg gggtggcggc cgccgtgcgc ccccaccgtc
     3481 cccctgcccg accccgcggt ctggccccag cccatgcctg cccacccccg acccgccccc
     3541 agagccgagc cccacgggct ggggaccgcc agacgggggt cgcgcggcgc ttgtgcgcag
     3601 ggctccgcag cccccgggcc gccccccgac gccggggccg cccctgtccg acgtctcccg
     3661 agtgtcgcgc cgcccagcct gggaggcgcg gtggccggtg cggaccgggc actgcgggag
     3721 gcacctctcg gcctccgagc ggcccctgtc gcccgcgcgc tgtcactaca gctcctttcc
     3781 tcgagccgac cgatccggcc gccccttcct cccgctcttc ccggagctgg aggacctgcc
     3841 gctgctcggt ccggagcagc tggcccggcg ggaggccctg ctgcacgcgg cctgggcccg
     3901 gggctcgcgc ccgcgtcacg cttccctgcc cagctccgtg gccgaggcct tcgctcggcc
     3961 cagctcgctg cccgctgggt gcaccggccc cgcctgcgcc cgccccgacg gccactcggc
     4021 ctgcaggcgc ttggcgcagg cgcagtcgat gtgcttgccg atctaccggg aggcctgcca
     4081 ggagggcgag caggcagggg cccccgcctg gcagcacaga cagcacgtct gcctgcacgc
     4141 ccacgcccac ctgccatttt gctggggggc tgtctgtcct caccttccac cctgtgccag
     4201 ccacggctcc tggctctccg gggcctgggg gcctctgggg cacaggggca ggactctggg
     4261 gctgggcaca ggctacagag acagtggggg actggacgag atcagcaggg tagcccgtgg
     4321 gacgcaaggc ttcccgggac cctgcacctg gagacggatc tccagtctgg agtcagaagt
     4381 gctcgaggac tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga
     4441 tgacgatgac aagtagtgag ggcccgatat ctcgacaatc aacctctgga ttacaaaatt
     4501 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     4561 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     4621 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     4681 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     4741 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     4801 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     4861 ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg
     4921 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     4981 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     5041 atctcccttt gggccgcctc cccgcctgga aacgggggag gctaactgaa acacggaagg
     5101 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     5161 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     5221 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     5281 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     5341 atagcagatc tgcgcagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     5401 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     5461 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     5521 caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg gcacctcgac
     5581 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     5641 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     5701 acaacactca accctatctc ggtctattct tttgatttat aagggatttt ggggatttcg
     5761 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     5821 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     5881 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     5941 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     6001 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     6061 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     6121 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     6181 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     6241 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     6301 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     6361 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     6421 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     6481 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     6541 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     6601 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     6661 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     6721 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     6781 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     6841 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     6901 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     6961 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     7021 tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     7081 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     7141 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     7201 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     7261 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     7321 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     7381 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     7441 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     7501 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     7561 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     7621 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     7681 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     7741 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     7801 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     7861 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     7921 gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt
     7981 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     8041 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     8101 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     8161 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     8221 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     8281 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     8341 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     8401 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     8461 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     8521 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     8581 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     8641 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     8701 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     8761 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     8821 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     8881 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     8941 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     9001 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     9061 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     9121 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     9181 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     9241 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     9301 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     9361 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     9421 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     9481 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     9541 ccctatggtc gactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
     9601 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
     9661 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag g
//
