LOCUS       V036783                 7388 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036783
VERSION     V036783
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             941..2878
                     /label="HSPA8(NM_006597)"
                     /note="HSPA8(NM_006597)"
                     /gene="HSPA8"
     CDS             2885..2950
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2988..3212
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3258..3686
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3700..4029
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4096..4890
                     /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    5064..5185
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5234..5250)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5258..5274
                     /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(5282..5312)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5327..5348
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5636..6221)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6392..7252)
                     /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(7253..7357)
                     /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 atccgccacc atgtccaagg gacctgcagt
      961 tggtattgat cttggcacca cctactcttg tgtgggtgtt ttccagcacg gaaaagtcga
     1021 gataattgcc aatgatcagg gaaaccgaac cactccaagc tatgtcgcct ttacggacac
     1081 tgaacggttg atcggtgatg ccgcaaagaa tcaagttgca atgaacccca ccaacacagt
     1141 ttttgatgcc aaacgtctga ttggacgcag atttgatgat gctgttgtcc agtctgatat
     1201 gaaacattgg ccctttatgg tggtgaatga tgctggcagg cccaaggtcc aagtagaata
     1261 caagggagag accaaaagct tctatccaga ggaggtgtct tctatggttc tgacaaagat
     1321 gaaggaaatt gcagaagcct accttgggaa gactgttacc aatgctgtgg tcacagtgcc
     1381 agcttacttt aatgactctc agcgtcaggc taccaaagat gctggaacta ttgctggtct
     1441 caatgtactt agaattatta atgagccaac tgctgctgct attgcttacg gcttagacaa
     1501 aaaggttgga gcagaaagaa acgtgctcat ctttgacctg ggaggtggca cttttgatgt
     1561 gtcaatcctc actattgagg atggaatctt tgaggtcaag tctacagctg gagacaccca
     1621 cttgggtgga gaagattttg acaaccgaat ggtcaaccat tttattgctg agtttaagcg
     1681 caagcataag aaggacatca gtgagaacaa gagagctgta agacgcctcc gtactgcttg
     1741 tgaacgtgct aagcgtaccc tctcttccag cacccaggcc agtattgaga tcgattctct
     1801 ctatgaagga atcgacttct atacctccat tacccgtgcc cgatttgaag aactgaatgc
     1861 tgacctgttc cgtggcaccc tggacccagt agagaaagcc cttcgagatg ccaaactaga
     1921 caagtcacag attcatgata ttgtcctggt tggtggttct actcgtatcc ccaagattca
     1981 gaagcttctc caagacttct tcaatggaaa agaactgaat aagagcatca accctgatga
     2041 agctgttgct tatggtgcag ctgtccaggc agccatcttg tctggagaca agtctgagaa
     2101 tgttcaagat ttgctgctct tggatgtcac tcctctttcc cttggtattg aaactgctgg
     2161 tggagtcatg actgtcctca tcaagcgtaa taccaccatt cctaccaagc agacacagac
     2221 cttcactacc tattctgaca accagcctgg tgtgcttatt caggtttatg aaggcgagcg
     2281 tgccatgaca aaggataaca acctgcttgg caagtttgaa ctcacaggca tacctcctgc
     2341 accccgaggt gttcctcaga ttgaagtcac ttttgacatt gatgccaatg gtatactcaa
     2401 tgtctctgct gtggacaaga gtacgggaaa agagaacaag attactatca ctaatgacaa
     2461 gggccgtttg agcaaggaag acattgaacg tatggtccag gaagctgaga agtacaaagc
     2521 tgaagatgag aagcagaggg acaaggtgtc atccaagaat tcacttgagt cctatgcctt
     2581 caacatgaaa gcaactgttg aagatgagaa acttcaaggc aagattaacg atgaggacaa
     2641 acagaagatt ctggacaagt gtaatgaaat tatcaactgg cttgataaga atcagactgc
     2701 tgagaaggaa gaatttgaac atcaacagaa agagctggag aaagtttgca accccatcat
     2761 caccaagctg taccagagtg caggaggcat gccaggagga atgcctgggg gatttcctgg
     2821 tggtggagct cctccctctg gtggtgcttc ctcagggccc accattgaag aggttgatct
     2881 cgaggactac aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga
     2941 cgatgacaag tagtgagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
     3001 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     3061 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     3121 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     3181 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     3241 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3301 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     3361 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     3421 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3481 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3541 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3601 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     3661 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     3721 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3781 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3841 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3901 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3961 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4021 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     4081 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     4141 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4201 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4261 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4321 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4381 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4441 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4501 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4561 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4621 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4681 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4741 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4801 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4861 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     4921 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     4981 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     5041 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     5101 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     5161 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     5221 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     5281 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     5341 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     5401 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     5461 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5521 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5581 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5641 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5701 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5761 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5821 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5881 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5941 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6001 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6061 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6121 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg
     6181 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6241 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6301 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     6361 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6421 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6481 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     6541 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6601 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6661 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     6721 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6781 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6841 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6901 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6961 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     7021 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7081 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7141 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7201 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     7261 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     7321 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     7381 ctgacgtc
//
