LOCUS       V036191                 8141 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036191
VERSION     V036191
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             908..3661
                     /label="GRIK1(NM_000830)"
                     /note="GRIK1(NM_000830)"
                     /gene="GRIK1"
     polyA_signal    3708..3932
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3978..4406
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4420..4749
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4816..5610
                     /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    5784..5905
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5954..5970)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5978..5994
                     /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(6002..6032)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6047..6068
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6356..6941)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7112..7972)
                     /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(7973..8077)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cgccaccatg gagcacggca cactcctcgc ccagcccggg ctctggacca gggacaccag
      961 ctgggcactc ctctatttcc tctgctatat cctccctcag accgccccgc aagtactcag
     1021 gatcggaggg atttttgaaa cagtggaaaa tgagcctgtt aatgttgaag aattagcttt
     1081 caagtttgca gtcaccagca ttaacagaaa ccgaaccctg atgcctaaca ccacattaac
     1141 ctatgacatc cagagaatta acctttttga tagttttgaa gcctcgcgga gagcatgtga
     1201 ccagctggct cttggtgtgg ctgctctctt tggcccttcc catagctcct ccgtcagtgc
     1261 tgtgcagtct atttgcaatg ctctcgaagt tccacacata cagacccgct ggaaacaccc
     1321 ctcggtggac aacaaagatt tgttttacat caacctttac ccagattatg cagctatcag
     1381 cagggcgatc ctggatctgg tcctctatta caactggaaa acagtgacag tggtgtatga
     1441 agacagcaca ggtctaattc gtctacaaga gctcatcaaa gctccctcca gatataatat
     1501 taaaatcaaa atccgccagc tgccctctgg gaataaagat gccaagcctt tactcaagga
     1561 gatgaagaaa ggcaaggagt tctatgtgat atttgattgt tcacatgaaa cagccgctga
     1621 aatccttaag cagattctgt tcatgggcat gatgaccgag tactatcact actttttcac
     1681 aaccctggac ttatttgctt tggatctgga actctatagg tacagtggcg taaacatgac
     1741 cgggtttcgg ctgcttaaca ttgacaaccc tcacgtgtca tccatcattg agaagtggtc
     1801 catggagaga ctgcaggccc cacccaggcc cgagactggc cttttggatg gcatgatgac
     1861 aactgaagcg gctctgatgt acgatgctgt gtacatggtg gccattgcct cgcaccgggc
     1921 atcccagctg accgtcagct ccctgcagtg ccatagacat aagccatggc gcctcggacc
     1981 cagatttatg aacctgatca aagaggcccg gtgggatggc ttgactgggc atatcacctt
     2041 taataaaacc aatggcttga ggaaggattt tgatctggac attattagtc tcaaagagga
     2101 aggaactgaa aaggctgctg gcgaagtgtc taaacacttg tataaagtgt ggaagaagat
     2161 tgggatttgg aattccaaca gtgggcttaa catgacggac agcaacaaag acaagtccag
     2221 caatatcact gattcattgg ccaacagaac actcattgtc accaccattc tggaagaacc
     2281 ctatgttatg tacaggaaat ctgataagcc tctatatgga aatgacagat ttgaaggata
     2341 ttgcctagac ctgttgaaag aattgtcaaa catcctgggt ttcatttatg atgttaaact
     2401 agttcccgat ggcaaatatg gggcccagaa tgacaaaggg gagtggaacg ggatggttaa
     2461 agaactcata gatcacaggg ctgacctggc agtggctcct cttaccatca cctacgtgcg
     2521 ggagaaagtc attgacttct ccaaaccctt catgacccta ggcatcagca ttctctaccg
     2581 gaagcccaat ggtaccaatc caggcgtttt ctccttcctc aaccccctgt ctccagatat
     2641 ttggatgtat gtgctcttag cctgcttggg agtcagctgt gtactctttg tgattgcaag
     2701 gtttacaccc tacgagtggt ataaccccca cccatgcaac cctgactcag acgtggtgga
     2761 aaacaatttt actttactaa atagtttctg gtttggagtt ggagctctca tgcagcaagg
     2821 atcagagctg atgcccaaag ctctatcgac cagaatagtt ggagggatat ggtggttttt
     2881 caccctaatc atcatttcat cctacacggc caatctggct gccttcttga cagtagagag
     2941 aatggaatcc cccatagatt cggcagatga tctggcaaag caaaccaaga tagaatatgg
     3001 ggcggttaga gatggatcaa caatgacctt cttcaagaaa tcaaaaatct ccacctatga
     3061 gaagatgtgg gctttcatga gcagcaggca gcagaccgcc ctggtaagaa acagtgatga
     3121 ggggatccag agagtgctca ccacagacta cgcgctgctg atggagtcca ccagcattga
     3181 gtatgtgacg cagagaaact gcaacctcac tcagatcggg ggcctcattg actccaaagg
     3241 ttacggagtg ggaacaccta ttggttctcc ttaccgggat aaaattacta ttgctattct
     3301 tcaactccaa gaagaaggga agctgcatat gatgaaagag aagtggtggc gtgggaatgg
     3361 ctgccccgag gaagacaaca aagaagccag tgccctggga gtggaaaata ttggaggcat
     3421 cttcattgtt ctggctgccg gactggtcct ttctgtattt gtagctattg gagaattcat
     3481 atacaaatca cggaagaata atgatattga acaggctttt tgtttctttt atggactgca
     3541 atgtaagcaa acccatccaa ccaactccac ttctggaact actttatcta cggatttaga
     3601 atgtggtaaa ttaattcgag aggagagagg gattcgaaaa cagtcctcag ttcatactgt
     3661 gtaactcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
     3721 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     3781 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     3841 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     3901 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     3961 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4021 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4081 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4141 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4201 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4261 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     4321 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     4381 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     4441 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4501 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4561 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4621 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4681 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4741 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     4801 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     4861 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4921 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4981 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     5041 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     5101 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     5161 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     5221 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     5281 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     5341 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     5401 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     5461 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     5521 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     5581 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     5641 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     5701 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     5761 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     5821 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     5881 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     5941 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     6001 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     6061 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     6121 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     6181 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     6241 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     6301 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     6361 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     6421 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     6481 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     6541 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     6601 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6661 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6721 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6781 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6841 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg
     6901 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6961 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     7021 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     7081 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     7141 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     7201 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     7261 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     7321 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     7381 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     7441 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     7501 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     7561 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     7621 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     7681 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     7741 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7801 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7861 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7921 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     7981 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     8041 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     8101 ctgacgtcga cggatcggga gatctcccga tcccctatgg t
//
