LOCUS       V018877                 8066 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018877
VERSION     V018877
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..2746
                     /label="GRK3(NM_005160)"
                     /note="GRK3(NM_005160)"
                     /gene="GRK3"
     CDS             2753..2818
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2842..3430
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3496..3544
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3746..4174
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4188..4517
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4584..5378
                     /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    5554..5675
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5724..5740)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5748..5764
                     /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(5772..5802)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5817..5838
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6126..6714)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6885..7745)
                     /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(7746..7850)
                     /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 ccatggcgga cctggaggct gtgctggccg atgtcagtta
      721 cctgatggcc atggagaaga gcaaggcgac cccggccgcc cgcgccagca agaggatcgt
      781 cctgccggag cccagtatcc ggagtgtgat gcagaagtac cttgcagaga gaaatgaaat
      841 aacctttgac aagattttca atcagaaaat tggtttcttg ctatttaaag atttttgttt
      901 gaatgaaatt aatgaagctg tacctcaggt gaagttttat gaagagataa aggaatatga
      961 aaaacttgat aatgaggaag accgcctttg cagaagtcga caaatttatg atgcctacat
     1021 catgaaggaa cttctttcct gttcacatcc tttctcaaag caagctgtag aacacgtaca
     1081 aagtcattta tccaagaaac aagtgacatc aactcttttt cagccataca tagaagaaat
     1141 ttgtgaaagc cttcgaggtg acatttttca aaaatttatg gaaagtgaca agttcactag
     1201 attttgtcag tggaaaaacg ttgaattaaa tatccatttg accatgaatg agttcagtgt
     1261 gcataggatt attggacgag gaggattcgg ggaagtttat ggttgcagga aagcagacac
     1321 tggaaaaatg tatgcaatga aatgcttaga taagaagagg atcaaaatga aacaaggaga
     1381 aacattagcc ttaaatgaaa gaatcatgtt gtctcttgtc agcacaggag actgtccttt
     1441 cattgtatgt atgacctatg ccttccatac cccagataaa ctctgcttca tcctggatct
     1501 gatgaacggg ggcgatttgc actaccacct ttcacaacac ggtgtgttct ctgagaagga
     1561 gatgcggttt tatgccactg aaatcattct gggtctggaa cacatgcaca atcggtttgt
     1621 tgtctacaga gatttgaagc cagcaaatat tctcttggat gaacatggac acgcaagaat
     1681 atcagatctt ggtcttgcct gcgatttttc caaaaagaag cctcatgcga gtgttggcac
     1741 ccatgggtac atggctcccg aggtgctgca gaaggggacg gcctatgaca gcagtgccga
     1801 ctggttctcc ctgggctgca tgcttttcaa acttctgaga ggtcacagcc ctttcagaca
     1861 acataaaacc aaagacaagc atgaaattga ccgaatgaca ctcaccgtga atgtggaact
     1921 tccagacacc ttctctcctg aactgaagtc ccttttggag ggcttgcttc agcgagacgt
     1981 tagcaagcgg ctgggctgtc acggaggcgg ctcacaggaa gtaaaagagc acagcttttt
     2041 caaaggtgtt gactggcagc atgtctactt acaaaagtac ccaccaccct tgattcctcc
     2101 ccggggagaa gtcaatgctg ctgatgcctt tgatattggc tcatttgatg aagaggatac
     2161 caaagggatt aagctacttg attgcgacca agaactctac aagaacttcc ctttggtcat
     2221 ctctgaacgc tggcagcaag aagtaacgga aacagtttat gaagcagtaa atgcagacac
     2281 agataaaatc gaggccagga agagagctaa aaataagcaa cttggccacg aagaagatta
     2341 cgctctgggg aaggactgta ttatgcacgg gtacatgctg aaactgggaa acccatttct
     2401 gactcagtgg cagcgtcgct atttttacct ctttccaaat agacttgaat ggagaggaga
     2461 gggagagtcc cggcaaaatt tactgacaat ggaacagatt ctctctgtgg aagaaactca
     2521 aattaaagac aaaaaatgca ttttgttcag aataaaagga gggaaacaat ttgtcttgca
     2581 atgtgagagt gatccagagt ttgtgcagtg gaagaaagag ttgaacgaaa ccttcaagga
     2641 ggcccagcgg ctattgcgtc gtgccccgaa gttcctcaac aaacctcggt caggtactgt
     2701 ggagctccca aagccatccc tctgtcacag aaacagcaac ggcctcctcg aggactacaa
     2761 agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta
     2821 gtgagggccc gatatctcga caatcaacct ctggattaca aaatttgtga aagattgact
     2881 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     2941 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     3001 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     3061 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     3121 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     3181 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg
     3241 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     3301 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     3361 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     3421 gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     3481 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt tgggtcgttt
     3541 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     3601 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     3661 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc agatctgcgc
     3721 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3781 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3841 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3901 ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3961 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4021 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     4081 atctcggtct attcttttga tttataaggg attttgggga tttcggccta ttggttaaaa
     4141 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     4201 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4261 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4321 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4381 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4441 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4501 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     4561 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4621 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4681 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4741 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     4801 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4861 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4921 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4981 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     5041 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     5101 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     5161 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     5221 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     5281 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     5341 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgcgaa
     5401 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     5461 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     5521 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     5581 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     5641 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     5701 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     5761 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     5821 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     5881 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     5941 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     6001 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     6061 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     6121 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     6181 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     6241 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     6301 ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     6361 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     6421 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     6481 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     6541 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     6601 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     6661 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     6721 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     6781 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     6841 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     6901 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     6961 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     7021 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     7081 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     7141 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     7201 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     7261 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     7321 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     7381 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     7441 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     7501 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     7561 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     7621 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     7681 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     7741 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     7801 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     7861 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtcgactc
     7921 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt
     7981 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
     8041 acaattgcat gaagaatctg cttagg
//
