LOCUS       V019370                 8942 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019370
VERSION     V019370
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..3622
                     /label="GRIK5(NM_002088)"
                     /note="GRIK5(NM_002088)"
                     /gene="GRIK5"
     CDS             3629..3694
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3718..4306
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4372..4420
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4622..5050
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5064..5393
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5460..6254
                     /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    6430..6551
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6600..6616)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6624..6640
                     /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(6648..6678)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6693..6714
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7002..7590)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7761..8621)
                     /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(8622..8726)
                     /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 ccatgccggc tgagctgctg ctgctgctga ttgttgcctt
      721 cgccagcccc agctgccagg tgctctcatc actgcgcatg gctgcaatcc tggatgatca
      781 gacagtgtgt ggccgcggtg agcgtctggc cttggccttg gcccgggagc agatcaacgg
      841 gatcatcgag gtcccagcca aggcccgagt ggaagtagac atctttgagc tgcagcggga
      901 cagccagtac gagaccacgg acaccatgtg tcagatctta cccaaagggg ttgtgtctgt
      961 ccttgggccc tcctctagcc cagcatctgc ctccaccgtg agccatatct gtggagagaa
     1021 ggagatcccc cacatcaagg tgggtcccga ggagacaccc cgccttcagt accttcgctt
     1081 cgcgtctgtc agcctgtacc ccagtaacga ggacgtcagc ttggcggtct cccgaatcct
     1141 caagtccttc aactacccct cggccagcct catctgcgcc aaggctgagt gcctgctgcg
     1201 attggaggaa ctggtgcgtg gcttcctcat ctccaaggag acgctgtcag tgaggatgtt
     1261 ggacgacagc cgggacccca caccactgct caaggagatc cgtgatgaca aggtgtccac
     1321 catcatcatc gacgccaacg cctccatctc ccacctcatc ctccgtaagg cctcggaact
     1381 gggaatgacc tcagcgtttt acaagtacat cctcaccacc atggacttcc ccatcctgca
     1441 tctggacggt attgtggagg actcctccaa catcctgggc ttctccatgt tcaacacgtc
     1501 ccaccccttc taccctgagt ttgtccgcag cctcaacatg tcctggaggg agaactgtga
     1561 agccagcacc tacctgggcc ctgcgctgtc agccgccctg atgtttgacg ccgtgcacgt
     1621 ggtggtgagc gctgtccgag agctgaaccg cagccaggag atcggtgtga agcctctggc
     1681 ctgtacatcg gccaacattt ggccccacgg gaccagcctc atgaactacc tgcgcatggt
     1741 agagtatgat gggctgaccg ggcgggtcga gttcaacagc aaagggcaga gaaccaacta
     1801 caccctgcgc atcctagaaa agtcccggca gggccaccgt gagattgggg tgtggtactc
     1861 taaccgcacc ctggccatga atgccaccac cctggacatc aacctgtcgc agacactggc
     1921 caacaagacc ctggtggtca caaccatcct ggagaaccca tacgtcatgc gccggcccaa
     1981 cttccaggcc ctgtcgggga acgaacgctt cgagggcttc tgcgtggaca tgctgcggga
     2041 gctggccgag ctgctgcgct tccgctaccg cctgcggttg gtggaggatg ggctgtacgg
     2101 ggcgcccgag cccaacggct cctggacggg catggttggc gagctcatca accggaaggc
     2161 agacctggct gtggccgcct tcaccatcac agctgagcgg gagaaggtca tcgacttttc
     2221 caagcccttt atgaccctgg ggatcagcat cctctaccga gtgcacatgg gccgcaagcc
     2281 tggctacttc tccttcctgg accccttctc ccctgctgtg tggctcttca tgcttcttgc
     2341 ctacctggct gtcagctgcg tcctgtttct ggctgccagg ctgagcccct atgagtggta
     2401 taacccacac ccatgcctgc gggcacgccc ccacatcctg gagaaccagt acacgctggg
     2461 caacagcctg tggtttcccg tggggggctt catgcagcag ggctcggaga tcatgccccg
     2521 ggcgctgtcc acgcgctgtg tcagcggagt ctggtgggcc ttcaccttga tcatcatctc
     2581 ctcctacacg gccaacctgg ccgccttcct caccgtgcag cgcatggagg tgcctgtgga
     2641 gtcggccgat gacctggcag atcagaccaa catcgagtat ggcaccatcc acgccggctc
     2701 caccatgacc ttcttccaga attcacggta ccaaacgtac cagcgcatgt ggaactacat
     2761 gcagtcgaag cagcccagcg tgttcgtcaa gagcacagaa gagggcattg cccgcgtcct
     2821 caactcccgc tacgccttcc tgctcgagtc caccatgaac gaataccacc ggcgcctcaa
     2881 ctgcaacctc acccagatcg ggggactcct cgacaccaag ggctacggca ttggcatgcc
     2941 gctgggctcc ccgttccggg atgagatcac actggccatc ctgcagcttc aggagaacaa
     3001 ccggctggag atcctgaagc gcaagtggtg ggaggggggc cggtgcccca aggaggagga
     3061 ccatcgagct aaaggtttgg gcatggagaa cattggtggc atttttatcg tgctcatctg
     3121 tggcctcatc attgctgtct tcgtggcggt catggaattc atatggtcca cacggaggtc
     3181 agctgagtcc gaggaggtgt cggtgtgcca ggagatgctg caggagctgc gccacgccgt
     3241 ttcttgccgc aagacgtcgc gttcccgccg gcgccgacgc ccgggcggcc cgagccgggc
     3301 cctgctgtca ctgcgcgcgg tccgcgagat gcgcctcagc aacggcaagc tctactcggc
     3361 cggcgcgggc ggggatgcgg gcagcgcgca cgggggcccg cagcgcctcc tggacgaccc
     3421 ggggcccccc agcggagccc gacccgccgc ccccaccccc tgcacccacg tgcgcgtctg
     3481 ccaggagtgc cggcgcatcc aggcgctgcg ggcctcgggg gccggcgcgc ctccgcgtgg
     3541 cctgggcgtc cccgccgaag ccaccagccc gccccggccg cggcctggcc ccgccggccc
     3601 ccgggagctg gcggagcacg agctcgagga ctacaaagac catgacggtg attataaaga
     3661 tcatgacatc gactacaagg atgacgatga caagtagtga gggcccgata tctcgacaat
     3721 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     3781 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     3841 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     3901 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     3961 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     4021 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     4081 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc
     4141 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     4201 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     4261 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg aaacggggga
     4321 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     4381 aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     4441 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     4501 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     4561 gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag ggggtatccc
     4621 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     4681 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4741 acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg gttccgattt
     4801 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     4861 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4921 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     4981 taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     5041 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     5101 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     5161 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     5221 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     5281 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     5341 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     5401 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     5461 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     5521 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     5581 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     5641 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     5701 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     5761 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     5821 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     5881 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     5941 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     6001 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     6061 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     6121 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     6181 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     6241 acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac
     6301 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     6361 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     6421 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     6481 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     6541 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     6601 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     6661 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     6721 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     6781 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     6841 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     6901 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     6961 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     7021 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     7081 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     7141 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc
     7201 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     7261 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     7321 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     7381 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     7441 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     7501 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     7561 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     7621 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     7681 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     7741 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     7801 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     7861 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     7921 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     7981 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     8041 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     8101 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     8161 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     8221 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     8281 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     8341 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     8401 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     8461 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     8521 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     8581 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     8641 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     8701 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     8761 ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc tctgatgccg
     8821 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
     8881 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
     8941 gg
//
