LOCUS       V040082                 6868 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040082
VERSION     V040082
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             964..2358
                     /label="G3bp1(NM_013716)"
                     /note="G3bp1(NM_013716)"
                     /gene="G3bp1"
     CDS             2365..2430
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2468..2692
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2738..3166
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3180..3509
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3576..4370
                     /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    4544..4665
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4714..4730)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4738..4754
                     /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(4762..4792)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4807..4828
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5116..5701)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5872..6732)
                     /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(6733..6837)
                     /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 atccactagt ccagtgtggt ggaattcgcc
      961 accatggtta tggagaagcc tagtcccctg ctggtcgggc gggagtttgt gcgacagtac
     1021 tacactctgc tgaaccaggc cccggacatg ttgcacagat tttatggaaa gaactcttcc
     1081 tatgcccacg ggggcttgga ttccaacggg aagccagcag atgcagtcta cgggcagaag
     1141 gaaatccaca ggaaagtgat gtcacaaaac ttcaccaact gccacaccaa gatccgccac
     1201 gtggatgctc acgcaactct gaatgacggg gtggtggtcc aagtgatggg gctactgtcc
     1261 aacaacaacc aggctctgcg gaggttcatg cagacctttg tccttgctcc tgagggctct
     1321 gttgccaaca aattctatgt tcacaacgac atcttcaggt accaagatga ggtcttcggt
     1381 ggctttgtca cagagcctca agaggaatcc gaggaagaag tagaagaacc tgaagaaaga
     1441 cagcagacac cagaggtggt gcctgatgat tctggaactt tctatgatca gactgtcagc
     1501 aatgacttgg aggagcattt agaggagcct gtagtggaac cagaaccgga gccagagccg
     1561 gagccggagc cggaacctgt gtccgacatt caagaggaca agcctgaggc tgcattggaa
     1621 gaagctgctc ccgacgatgt gcagaagagc acttccccgg ccccggctga tgtggccccg
     1681 gcacaggagg acttgaggac gttttcttgg gcatctgtga cgagtaagaa ccttcctccc
     1741 agtggggctg ttccagtgac ggggacacca cctcatgtgg tcaaagtgcc agcgtcacag
     1801 ccccgtccag aatctaaacc tgactctcag atcccaccac aaaggcctca gagagatcag
     1861 agagttcgag agcagcgaat caatatccct cctcagagag gaccccgacc aatccgtgag
     1921 gctggtgaac caggagatgt ggaacctcga aggatggtga ggcaccctga cagtcaccag
     1981 ctcttcattg ggaacctgcc acatgaggtt gacaagtcgg aactgaagga ttttttccaa
     2041 aattttggca atgtggtgga gctgcgcatc aacagtggtg ggaagttacc caatttcggt
     2101 tttgttgtgt ttgatgattc tgaacctgtt cagaaggtcc ttagtaatag gcccatcatg
     2161 ttccgaggcg cggtccgcct gaatgtggaa gagaagaaga ctcgagctgc cagagaaggc
     2221 gaccgcaggg ataaccgcct tcggggacct ggaggccccc gtggtgggcc gagtggtggg
     2281 atgagaggcc ctccccgcgg aggcatggtg cagaaaccag gctttggcgt gggcaggggg
     2341 attacaactc caaggcagct cgaggactac aaagaccatg acggtgatta taaagatcat
     2401 gacatcgact acaaggatga cgatgacaag tagtgagggc ccgtttaaac ccgctgatca
     2461 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     2521 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     2581 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     2641 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     2701 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     2761 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2821 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2881 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2941 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3001 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3061 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     3121 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     3181 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3241 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3301 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3361 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3421 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     3481 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     3541 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     3601 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     3661 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     3721 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     3781 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     3841 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     3901 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     3961 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     4021 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     4081 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     4141 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     4201 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     4261 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     4321 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     4381 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     4441 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     4501 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     4561 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4621 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     4681 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     4741 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4801 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4861 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4921 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4981 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5041 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5101 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5161 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5221 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5281 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5341 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5401 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5461 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5521 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5581 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5641 cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5701 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5761 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5821 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5881 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5941 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6001 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6061 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6121 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6181 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6241 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6301 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6361 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6421 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6481 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6541 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6601 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6661 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6721 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6781 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6841 atttccccga aaagtgccac ctgacgtc
//
