LOCUS       V024781                 8066 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024781
VERSION     V024781
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             941..3556
                     /label="GRM2(NM_000839)"
                     /note="GRM2(NM_000839)"
                     /gene="GRM2"
     CDS             3563..3628
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3666..3890
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3936..4364
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4378..4707
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4774..5568
                     /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    5742..5863
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5912..5928)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5936..5952
                     /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(5960..5990)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6005..6026
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6314..6899)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7070..7930)
                     /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(7931..8035)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgggatcgc tgcttgcgct
      961 cctggcactg ctgctgctgt ggggtgctgt ggctgagggc ccagccaaga aggtgctgac
     1021 cctggaggga gacttggtgc tgggtgggct gttcccagtg caccagaagg gcggcccagc
     1081 agaggactgt ggtcctgtca atgagcaccg tggcatccag cgcctggagg ccatgctttt
     1141 tgcactggac cgcatcaacc gtgacccgca cctgctgcct ggcgtgcgcc tgggtgcaca
     1201 catcctcgac agttgctcca aggacacaca tgcgctggag caggcactgg actttgtgcg
     1261 tgcctcactc agccgtggtg ctgatggctc acgccacatc tgccccgacg gctcttatgc
     1321 gacccatggt gatgctccca ctgccatcac tggtgttatt ggcggttcct acagtgatgt
     1381 ctccatccag gtggccaacc tcttgaggct atttcagatc ccacagatta gctacgcctc
     1441 taccagtgcc aagctgagtg acaagtcccg ctatgactac tttgcccgca cagtgcctcc
     1501 tgacttcttc caagccaagg ccatggctga gattctccgc ttcttcaact ggacctatgt
     1561 gtccactgtg gcgtctgagg gcgactatgg cgagacaggc attgaggcct ttgagctaga
     1621 ggctcgtgcc cgcaacatct gtgtggccac ctcggagaaa gtgggccgtg ccatgagccg
     1681 cgcggccttt gagggtgtgg tgcgagccct gctgcagaag cccagtgccc gcgtggctgt
     1741 cctgttcacc cgttctgagg atgcccggga gctgcttgct gccagccagc gcctcaatgc
     1801 cagcttcacc tgggtggcca gtgatggttg gggggccctg gagagtgtgg tggcaggcag
     1861 tgagggggct gctgagggtg ctatcaccat cgagctggcc tcctacccca tcagtgactt
     1921 tgcctcctac ttccagagcc tggacccttg gaacaacagc cggaacccct ggttccgtga
     1981 attctgggag cagaggttcc gctgcagctt ccggcagcga gactgcgcag cccactctct
     2041 ccgggctgtg ccctttgagc aggagtccaa gatcatgttt gtggtcaatg cagtgtacgc
     2101 catggcccat gcgctccaca acatgcaccg tgccctctgc cccaacacca cccggctctg
     2161 tgacgcgatg cggccagtta acgggcgccg cctctacaag gactttgtgc tcaacgtcaa
     2221 gtttgatgcc ccctttcgcc cagctgacac ccacaatgag gtccgctttg accgctttgg
     2281 tgatggtatt ggccgctaca acatcttcac ctatctgcgt gcaggcagtg ggcgctatcg
     2341 ctaccagaag gtgggctact gggcagaagg cttgactctg gacaccagcc tcatcccatg
     2401 ggcctcaccc tcagccggcc ccctgcccgc ctctcgctgc agtgagccct gcctccagaa
     2461 tgaggtgaag agtgtgcagc cgggcgaagt ctgctgctgg ctctgcattc cgtgccagcc
     2521 ctatgagtac cgattggacg aattcacttg cgctgattgt ggcctgggct actggcccaa
     2581 tgccagcctg actggctgct tcgaactgcc ccaggagtac atccgctggg gcgatgcctg
     2641 ggctgtggga cctgtcacca tcgcctgcct cggtgccctg gccaccctct ttgtgctggg
     2701 tgtctttgtg cggcacaatg ccacaccagt ggtcaaggcc tcaggtcggg agctctgcta
     2761 catcctgctg ggtggtgtct tcctctgcta ctgcatgacc ttcatcttca ttgccaagcc
     2821 atccacggca gtgtgtacct tacggcgtct tggtttgggc actgccttct ctgtctgcta
     2881 ctcagccctg ctcaccaaga ccaaccgcat tgcacgcatc ttcggtgggg cccgggaggg
     2941 tgcccagcgg ccacgcttca tcagtcctgc ctcacaggtg gccatctgcc tggcacttat
     3001 ctcgggccag ctgctcatcg tggtcgcctg gctggtggtg gaggcaccgg gcacaggcaa
     3061 ggagacagcc cccgaacggc gggaggtggt gacactgcgc tgcaaccacc gcgatgcaag
     3121 tatgttgggc tcgctggcct acaatgtgct cctcatcgcg ctctgcacgc tttatgcctt
     3181 caagactcgc aagtgccccg aaaacttcaa cgaggccaag ttcattggct tcaccatgta
     3241 caccacctgc atcatctggc tggcattcct gcccatcttc tatgtcacct ccagtgacta
     3301 ccgggtacag accaccacca tgtgcgtgtc agtcagcctc agcggctccg tggtgcttgg
     3361 ctgcctcttt gcgcccaagc tgcacatcat cctcttccag ccgcagaaga acgtggttag
     3421 ccaccgggca cccaccagcc gctttggcag tgctgctgcc agggccagct ccagccttgg
     3481 ccaagggtct ggctcccagt ttgtccccac tgtttgcaat ggccgtgagg tggtggactc
     3541 gacaacgtca tcgcttctcg aggactacaa agaccatgac ggtgattata aagatcatga
     3601 catcgactac aaggatgacg atgacaagta gtgagggccc gtttaaaccc gctgatcagc
     3661 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     3721 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     3781 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     3841 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     3901 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     3961 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     4021 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4081 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4141 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4201 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4261 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     4321 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     4381 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4441 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     4501 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     4561 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4621 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     4681 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     4741 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     4801 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     4861 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     4921 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     4981 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     5041 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     5101 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     5161 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     5221 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     5281 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     5341 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     5401 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     5461 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     5521 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     5581 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     5641 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     5701 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     5761 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     5821 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     5881 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     5941 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     6001 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6061 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6121 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6181 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6241 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6301 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6361 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6421 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6481 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     6541 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6601 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6661 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     6721 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     6781 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     6841 ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6901 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6961 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     7021 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     7081 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     7141 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     7201 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7261 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7321 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7381 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7441 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7501 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7561 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     7621 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7681 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7741 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7801 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7861 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7921 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7981 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     8041 ttccccgaaa agtgccacct gacgtc
//
