LOCUS       V034465                 7667 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034465
VERSION     V034465
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             884..1591
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
                     /label="mCherry"
     CDS             1628..3025
                     /label="G3BP1(NM_005754)"
                     /note="G3BP1(NM_005754)"
                     /gene="G3BP1"
     CDS             3026..3106
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /transl_table=1
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
                     /label="3xHA"
     CDS             3107..3187
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /transl_table=1
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
                     /label="3xHA"
     polyA_signal    3234..3458
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3504..3932
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3946..4275
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4342..5136
                     /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    5310..5431
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5480..5496)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5504..5520
                     /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(5528..5558)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5573..5594
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5882..6467)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6638..7498)
                     /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(7499..7603)
                     /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 ggctagcgct accggtcgcc accatggtga gcaagggcga
      901 ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc
      961 cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
     1021 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct
     1081 gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga
     1141 ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
     1201 cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa
     1261 ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat
     1321 gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat
     1381 caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta
     1441 caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat
     1501 cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca
     1561 ctccaccggc ggcatggacg agctgtacaa gggtggaggc ggatcaggag gcggtggatc
     1621 tctcgagatg gtgatggaga agcctagtcc cctgctggtc gggcgggaat ttgtgagaca
     1681 gtattacaca ctgctgaacc aggccccaga catgctgcat agattttatg gaaagaactc
     1741 ttcttatgtc catgggggat tggattcaaa tggaaagcca gcagatgcag tctacggaca
     1801 gaaagaaatc cacaggaaag tgatgtcaca aaacttcacc aactgccaca ccaagattcg
     1861 ccatgttgat gctcatgcca cgctaaatga tggtgtggta gtccaggtga tggggcttct
     1921 ctctaacaac aaccaggctt tgaggagatt catgcaaacg tttgtccttg ctcctgaggg
     1981 gtctgttgca aataaattct atgttcacaa tgatatcttc agataccaag atgaggtctt
     2041 tggtgggttt gtcactgagc ctcaggagga gtctgaagaa gaagtagagg aacctgaaga
     2101 aagacagcaa acacctgagg tggtacctga tgattctgga actttctatg atcaggcagt
     2161 tgtcagtaat gacatggaag aacatttaga ggagcctgtt gctgaaccag agcctgatcc
     2221 tgaaccagaa ccagaacaag aacctgtatc tgaaatccaa gaggaaaagc ctgagccagt
     2281 attagaagaa actgcccctg aggatgctca gaagagttct tctccagcac ctgcagacat
     2341 agctcagaca gtacaggaag acttgaggac attttcttgg gcatctgtga ccagtaagaa
     2401 tcttccaccc agtggagctg ttccagttac tgggatacca cctcatgttg ttaaagtacc
     2461 agcttcacag ccccgtccag agtctaagcc tgaatctcag attccaccac aaagacctca
     2521 gcgggatcaa agagtgcgag aacaacgaat aaatattcct ccccaaaggg gacccagacc
     2581 aatccgtgag gctggtgagc aaggtgacat tgaaccccga agaatggtga gacaccctga
     2641 cagtcaccaa ctcttcattg gcaacctgcc tcatgaagtg gacaaatcag agcttaaaga
     2701 tttctttcaa agttatggaa acgtggtgga gttgcgcatt aacagtggtg ggaaattacc
     2761 caattttggt tttgttgtgt ttgatgattc tgagcctgtt cagaaagtcc ttagcaacag
     2821 gcccatcatg ttcagaggtg aggtccgtct gaatgtcgaa gagaagaaga ctcgagctgc
     2881 cagggaaggc gaccgacgag ataatcgcct tcggggacct ggaggccctc gaggtgggct
     2941 gggtggtgga atgagaggcc ctccccgtgg aggcatggtg cagaaaccag gatttggagt
     3001 gggaaggggg cttgcgccac ggcagtatcc gtacgatgtt ccagattacg cttatcccta
     3061 cgacgtgcct gattatgcat acccatatga tgtccccgac tatgcctacc cttacgacgt
     3121 gccagactac gcctacccat acgatgtgcc tgactacgct tatccatacg acgtgccgga
     3181 ctacgcctaa ggatcctcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     3241 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3301 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3361 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3421 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     3481 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3541 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3601 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3661 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3721 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3781 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3841 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     3901 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     3961 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4021 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4081 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4141 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4201 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4261 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     4321 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     4381 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4441 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4501 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     4561 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4621 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4681 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4741 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4801 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4861 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4921 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4981 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     5041 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     5101 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     5161 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     5221 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     5281 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     5341 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     5401 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     5461 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     5521 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     5581 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     5641 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     5701 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5761 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5821 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5881 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5941 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6001 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6061 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     6121 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6181 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6241 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6301 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     6361 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt
     6421 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6481 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6541 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6601 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6661 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6721 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6781 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6841 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6901 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6961 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     7021 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     7081 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     7141 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     7201 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     7261 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     7321 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     7381 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     7441 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     7501 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7561 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     7621 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggt
//
