LOCUS       V031118                 9287 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031118
VERSION     V031118
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     primer_bind     166..182
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     enhancer        343..722
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        723..926
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        952..970
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             1004..5449
                     /label="Grin2b(NM_008171)"
                     /note="Grin2b(NM_008171)"
                     /gene="Grin2b"
     CDS             5453..5482
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             5501..5524
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(5544..5560)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    5571..6193
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(6342..6930)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    7259..7306
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(7538..8332)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     promoter        complement(8367..8724)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(8726..8830)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      8857..9285
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
ORIGIN
        1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
       61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
      241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
      301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
      361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac
      961 tcactatagg gcggccggga attcgtcgac tggatccgcc accatgaagc ccagcgcaga
     1021 gtgctgttct cccaagttct ggttggtgtt ggccgtcttg gccgtatcgg gcagcaaagc
     1081 tcgttcccaa aagagcgccc ccagcatcgg catcgctgtc atcctcgtgg gcacttccga
     1141 cgaagtggcc ataaaagatg cccacgagaa agatgacttc catcatctct cagtagttcc
     1201 ccgggtggag ctggtagcca tgaacgagac tgacccaaag agcataatca cccgcatctg
     1261 cgatcttatg tctgaccgga agatccaggg ggtggtgttc gcggatgaca cggaccagga
     1321 agccatcgcc cagatcctcg atttcatttc tgctcagact ctcaccccca tcctgggcat
     1381 ccatgggggc tcatctatga taatggcaga taaggatgag tcctccatgt tcttccagtt
     1441 tggcccatcc attgaacagc aagcttctgt catgctcaac atcatggaag aatacgactg
     1501 gtacatcttc tccatcgtca ccacctactt ccccggctac caggacttcg tgaacaagat
     1561 ccgcagcact attgagaaca gctttgtggg ctgggagctc gaggaagtcc tcctgctaga
     1621 catgtctcta gacgatggcg actctaagat ccagaatcag ctgaagaagc tgcagagccc
     1681 catcattctc ctctactgca caaaggaaga agccacctac atcttcgaag tagctaactc
     1741 agttgggctg actggctacg gctacacatg gatcgtgccg agtctggtgg cgggggatac
     1801 ggacacggtg ccttcagagt tccccacggg gctcatctct gtgtcatatg acgaatggga
     1861 ctatggcctt cctgccagag tgagagatgg gattgccatc atcaccactg ctgcctcgga
     1921 catgctgtcc gaacacagtt tcatccctga gcccaagagc agttgctaca acacccacga
     1981 gaagaggatc taccagtcta acatgctgaa taggtatctg atcaacgtca cttttgaagg
     2041 gagaaacctg tccttcagtg aagatggcta ccagatgcat ccgaagctgg tgataatcct
     2101 tctgaacaag gagaggaagt gggagagggt gggaaaatgg aaagacaagt ccctgcagat
     2161 gaaatactat gtgtggcctc gaatgtgtcc tgagactgaa gaacaggaag atgaccatct
     2221 gagcatcgtt accttggagg aggcaccgtt tgtcattgtg gaaagtgtgg accctctcag
     2281 tgggacctgc atgcggaata cagtcccgtg ccagaagcgc atcatctctg agaataaaac
     2341 agatgaggaa ccaggctaca tcaaaaaatg ctgcaagggg ttttgtattg atatccttaa
     2401 gaaaatttct aagtctgtga agttcaccta tgacctttac ctggtgacca atggcaagca
     2461 tggaaagaaa atcaacggga cctggaacgg catgattggt gaggtggtca tgaagagggc
     2521 ctatatggca gtgggatcac taactatcaa tgaagaacgg tcagaggtgg ttgacttctc
     2581 tgtgcccttc atagagactg gcatcagtgt catggtatca cgcagcaatg ggactgtgtc
     2641 accttctgcc ttcttagagc cattcagtgc tgacgtgtgg gtgatgatgt ttgtgatgct
     2701 gctcattgtc tctgctgtag ctgtctttgt ctttgaatac ttcagccctg tgggttacaa
     2761 ccggtgccta gctgatggca gagagccagg cggcccatct ttcaccatcg gcaaagcgat
     2821 ttggttactc tggggtctgg tgtttaacaa ctccgtacct gtgcagaacc caaaggggac
     2881 cacctccaag atcatggtgt cagtgtgggc cttctttgct gtcattttcc tggccagcta
     2941 cactgccaac ttagccgcct tcatgatcca agaggagtat gtggaccagg tttccggcct
     3001 gagtgacaag aagttccaga gacctaatga cttctcaccc cctttccgct ttgggactgt
     3061 gcccaatggc agcacagaga ggaatatccg taataactat gcagaaatgc atgcctacat
     3121 gggaaagttc aaccaaaggg gtgtagatga tgccttgctc tccctgaaaa cagggaaact
     3181 tgatgcattc atctacgatg cagccgtgct caactacatg gctggaagag acgaaggctg
     3241 caagctggtg accattggca gtggcaaggt ctttgcttct acgggctatg gcattgctat
     3301 ccaaaaagac tctggttgga aacgccaggt ggaccttgct atcctgcagc tgtttggaga
     3361 tggggagatg gaagaactgg aagctctctg gctcactggc atttgccaca atgagaagaa
     3421 tgaggttatg agcagccagc tggacattga caacatggcg ggcgtcttct atatgttggg
     3481 ggcagccatg gctctcagcc tcatcacctt catctgtgaa catctcttct attggcagtt
     3541 ccgacattgc ttcatgggtg tctgttctgg caagcctggc atggtcttct ccatcagcag
     3601 aggtatctac agctgtatcc acggagtagc tatagaggag cgccaatccg tgatgaactc
     3661 ccccactgcc accatgaaca acacacactc caatatccta cgcttgctcc gaacggccaa
     3721 aaacatggcc aacctgtctg gagtcaacgg ctccccccag agtgccctgg acttcatccg
     3781 ccgtgagtcc tctgtctatg acatctctga gcatcgccgc agcttcacgc attcagactg
     3841 caagtcgtac aataacccac cctgtgagga aaacctgttc agtgactaca ttagtgaggt
     3901 agagagaaca tttggcaacc tgcagctgaa ggacagcaat gtgtaccaag accactatca
     3961 ccatcaccac cggccccaca gcatcggcag caccagctcc attgatgggc tctatgactg
     4021 tgacaaccca ccctttacca cccagcccag gtcaatcagc aagaaacccc tggacattgg
     4081 cctgccctcc tccaaacaca gccagctcag cgacctgtac ggcaagttct ctttcaagag
     4141 tgaccgctac agtggccatg atgacttgat tcgatcagat gtctcagaca tctccacgca
     4201 tactgtcacc tatggcaaca tcgagggcaa cgcagccaag aggaggaagc agcaatataa
     4261 ggacagtcta aagaagcggc cagcctcggc caaatctagg agggagtttg atgaaatcga
     4321 gctggcctac cgtcgccgac caccccgctc cccagaccac aagcgctact tcagggacaa
     4381 agaagggctc cgagacttct acctggacca gttccgaaca aaggagaact cgcctcactg
     4441 ggagcacgtg gacttgactg acatttacaa agaacgtagt gatgacttca agcgagattc
     4501 ggtcagtgga ggcgggccct gtaccaacag gtctcacctt aaacacggaa caggcgataa
     4561 gcacggagtg gtaggcgggg tgcctgctcc ttgggagaag aacctgacca atgtggattg
     4621 ggaggatagg tctgggggca acttctgccg cagctgtccc tccaagctgc acaattactc
     4681 ctctacggtg gcagggcaaa actcgggccg gcaggcctgc atcaggtgtg aggcctgcaa
     4741 gaaggctggc aacctgtatg acatcagcga ggacaactcc ctgcaggaac tggaccagcc
     4801 tgctgcccct gtggctgtgt catccaacgc ctccaccacc aagtatcctc aaagcccgac
     4861 taattccaag gcccagaaga agaatcggaa caaactgcgc cggcagcact cctacgacac
     4921 cttcgtggac ctgcagaagg aggaggccgc cttggcccca cgcagcgtga gcctgaaaga
     4981 caagggccga ttcatggatg ggagccccta cgcccatatg tttgagatgc cagctggtga
     5041 gagctccttt gccaacaagt cctcagtgac cactgccgga caccatcaca acaatcccgg
     5101 cagcggctac atgctcagca agtcgctcta ccctgaccgg gtcacgcaaa accctttcat
     5161 ccccactttt ggggatgatc agtgcttgct tcacggcagc aaatcctact tcttcaggca
     5221 gcccacggtg gcaggggcgt cgaaaacaag gccggacttc cgggcccttg tcaccaataa
     5281 gccagtggtg tcagcccttc atggggctgt gccaggtcgt ttccagaagg acatttgtat
     5341 agggaaccag tccaacccct gtgtgcctaa caacaaaaac cccagggctt tcaatggctc
     5401 cagcaatgga catgtttatg agaaactttc tagtattgag tctgatgtcc tcgagcagaa
     5461 actcatctca gaagaggatc tggcagcaaa tgatatcctg gattacaagg atgacgacga
     5521 taaggtttaa acggccggcc gcggtcatag ctgtttcctg aacagatccc gggtggcatc
     5581 cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca
     5641 gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata
     5701 ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg
     5761 cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa
     5821 tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc
     5881 aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca
     5941 tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca
     6001 aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg attttaaaat
     6061 aactatacca gcaggaggac gtccagacac agcataggct acctggccat gcccaaccgg
     6121 tgggacattt gagttgcttg cttggcactg tcctctcatg cgttgggtcc actcagtaga
     6181 tgcctgttga attgggtacg cggccagcgg cgagcggtat cagctcactc aaaggcggta
     6241 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     6301 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     6361 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     6421 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6481 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     6541 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6601 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6661 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6721 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6781 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6841 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6901 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6961 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     7021 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     7081 gagtaacctg aggctatggc agggcctgcc gccccgacgt tggctgcgag ccctgggcct
     7141 tcacccgaac ttggggggtg gggtggggaa aaggaagaaa cgcgggcgta ttggccccaa
     7201 tggggtctcg gtggggtatc gacagagtgc cagccctggg accgaacccc gcgtttatga
     7261 acaaacgacc caacaccgtg cgttttattc tgtcttttta ttgccgtcat agcgcgggtt
     7321 ccttccggta ttgtctcctt ccgtgtttca gttagcctcc ccctagggtg ggcgaagaac
     7381 tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa aacgattccg
     7441 aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc
     7501 gtcgcttggt cggtcatttc gaaccccaga gtcccgctca gaagaactcg tcaagaaggc
     7561 gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt
     7621 cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat
     7681 agcgatccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca
     7741 ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg ccgtcgggca
     7801 tgctcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca
     7861 gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt
     7921 tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat
     7981 cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg
     8041 gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg
     8101 cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct tgcagttcat
     8161 tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc
     8221 ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc
     8281 tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg
     8341 atcctcatcc tgtctcttga tcgatctttg caaaagccta ggcctccaaa aaagcctcct
     8401 cactacttct ggaatagctc agaggccgag gcggcctcgg cctctgcata aataaaaaaa
     8461 attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg cgggatgggc
     8521 ggagttaggg gcgggactat ggttgctgac taattgagat gcatgctttg catacttctg
     8581 cctgctgggg agcctgggga ctttccacac ctggttgctg actaattgag atgcatgctt
     8641 tgcatacttc tgcctgctgg ggagcctggg gactttccac accctaactg acacacattc
     8701 cacagctggt tctttccgcc tcaggactct tcctttttca atattattga agcatttatc
     8761 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     8821 gggttccgcg cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa
     8881 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     8941 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     9001 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     9061 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     9121 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     9181 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     9241 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaatttt
//
