LOCUS       V031371                 8138 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031371
VERSION     V031371
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..2818
                     /label="TICAM1(NM_182919)"
                     /note="TICAM1(NM_182919)"
                     /gene="TICAM1"
     CDS             2825..2890
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2914..3502
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3568..3616
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3818..4246
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4260..4589
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4656..5450
                     /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    5626..5747
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5796..5812)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5820..5836
                     /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(5844..5874)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5889..5910
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6198..6786)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6957..7817)
                     /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(7818..7922)
                     /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 ccatggcctg cacaggccca tcacttccta gcgccttcga
      721 cattctaggt gcagcaggcc aggacaagct cttgtatctg aagcacaaac tgaagacccc
      781 acgcccaggc tgccaggggc aggacctcct gcatgccatg gttctcctga agctgggcca
      841 ggaaactgag gccaggatct ctctagaggc attgaaggcc gatgcggtgg cccggctggt
      901 ggcccgccag tgggctggcg tggacagcac cgaggaccca gaggagcccc cagatgtgtc
      961 ctgggctgtg gcccgcttgt accacctgct ggctgaggag aagctgtgcc ccgcctcgct
     1021 gcgggacgtg gcctaccagg aagccgtccg caccctcagc tccagggacg accaccggct
     1081 gggggaactt caggatgagg cccgaaaccg gtgtgggtgg gacattgctg gggatccagg
     1141 gagcatccgg acgctccagt ccaatctggg ctgcctccca ccatcctcgg ctttgccctc
     1201 tgggaccagg agcctcccac gccccattga cggtgtttcg gactggagcc aagggtgctc
     1261 cctgcgatcc actggcagcc ctgcctccct ggccagcaac ttggaaatca gccagtcccc
     1321 taccatgccc ttcctcagcc tgcaccgcag cccacatggg cccagcaagc tctgtgacga
     1381 cccccaggcc agcttggtgc ccgagcctgt ccccggtggc tgccaggagc ctgaggagat
     1441 gagctggccg ccatcggggg agattgccag cccaccagag ctgccaagca gcccacctcc
     1501 tgggcttccc gaagtggccc cagatgcaac ctccactggc ctccctgata cccccgcagc
     1561 tccagaaacc agcaccaact acccagtgga gtgcaccgag gggtctgcag gcccccagtc
     1621 tctccccttg cctattctgg agccggtcaa aaacccctgc tctgtcaaag accagacgcc
     1681 actccaactt tctgtagaag ataccacctc tccaaatacc aagccgtgcc cacctactcc
     1741 caccacccca gaaacatccc ctcctcctcc tcctcctcct ccttcatcta ctccttgttc
     1801 agctcacctg accccctcct ccctgttccc ttcctccctg gaatcatcat cggaacagaa
     1861 attctataac tttgtgatcc tccacgccag ggcagacgaa cacatcgccc tgcgggttcg
     1921 ggagaagctg gaggcccttg gcgtgcccga cggggccacc ttctgcgagg atttccaggt
     1981 gccggggcgc ggggagctga gctgcctgca ggacgccata gaccactcag ctttcatcat
     2041 cctacttctc acctccaact tcgactgtcg cctgagcctg caccaggtga accaagccat
     2101 gatgagcaac ctcacgcgac aggggtcgcc agactgtgtc atccccttcc tgcccctgga
     2161 gagctccccg gcccagctca gctccgacac ggccagcctg ctctccgggc tggtgcggct
     2221 ggacgaacac tcccagatct tcgccaggaa ggtggccaac accttcaagc cccacaggct
     2281 tcaggcccga aaggccatgt ggaggaagga acaggacacc cgagccctgc gggaacagag
     2341 ccaacacctg gacggtgagc ggatgcaggc ggcggcactg aacgcagcct actcagccta
     2401 cctccagagc tacttgtcct accaggcaca gatggagcag ctccaggtgg cttttgggag
     2461 ccacatgtca tttgggactg gggcgcccta tggggctcga atgccctttg ggggccaggt
     2521 gcccctggga gccccgccac cctttcccac ttggccgggg tgcccgcagc cgccacccct
     2581 gcacgcatgg caggctggca cccccccacc gccctcccca cagccagcag cctttccaca
     2641 gtcactgccc ttcccgcagt ccccagcctt ccctacggcc tcacccgcac cccctcagag
     2701 cccagggctg caacccctca ttatccacca cgcacagatg gtacagctgg ggctgaacaa
     2761 ccacatgtgg aaccagagag ggtcccaggc gcccgaggac aagacgcagg aggcagaact
     2821 cgaggactac aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga
     2881 cgatgacaag tagtgagggc ccgatatctc gacaatcaac ctctggatta caaaatttgt
     2941 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     3001 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     3061 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     3121 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     3181 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     3241 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     3301 tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc
     3361 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     3421 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     3481 tccctttggg ccgcctcccc gcctggaaac gggggaggct aactgaaaca cggaaggaga
     3541 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacgggt
     3601 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     3661 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     3721 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     3781 gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     3841 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     3901 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3961 gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4021 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4081 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4141 acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg gatttcggcc
     4201 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     4261 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4321 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4381 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4441 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4501 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     4561 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     4621 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     4681 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     4741 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     4801 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     4861 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     4921 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     4981 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     5041 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     5101 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     5161 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     5221 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     5281 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     5341 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     5401 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     5461 ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     5521 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     5581 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     5641 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5701 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     5761 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5821 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5881 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5941 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6001 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6061 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6121 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6181 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6241 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6301 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6361 tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg
     6421 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6481 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6541 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6601 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     6661 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6721 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6781 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6841 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6901 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6961 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     7021 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     7081 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7141 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7201 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7261 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7321 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7381 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7441 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7501 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7561 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7621 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7681 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7741 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7801 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7861 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7921 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc
     7981 tatggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     8041 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
     8101 aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
