LOCUS       V047245                 8829 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047245
VERSION     V047245
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..3509
                     /label="Gba2(NM_172692)"
                     /note="Gba2(NM_172692)"
                     /gene="Gba2"
     CDS             3516..3581
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3605..4193
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4259..4307
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4509..4937
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4951..5280
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5347..6141
                     /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    6317..6438
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6487..6503)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6511..6527
                     /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(6535..6565)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6580..6601
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6889..7477)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7648..8508)
                     /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(8509..8613)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggt aacctgcgtc ccggcctcag
      781 agcaggtcgg ctgtgccgaa agagattccc aagtttattg tgaagatact ggcggcactg
      841 aggctgtgcg ggttacagac tgcgggagcc ccgaggatag tggaccccag gatgaaccaa
      901 gctactgcaa ttcagaggac tctgggcagc tgatggcctc ctacgagggt aaagctaggg
      961 gctaccaggt gcctcctttt ggctggcgga tctgcttggc tcacgagttt gcagagaaga
     1021 ggagaccttt tcaagctaac aacatctctc tcagtaattt ggtaaagcac ctcggtatgg
     1081 gcttgaggta cttgaagtgg tggtaccgaa agacccacgt ggaaaagaag acccctttca
     1141 tcgacatgct caattctcta cccctgagac agatctatgg ttgtcccctg ggtggcattg
     1201 gaggaggcac tatcacccgg ggctggagag gccagttctg tcgttggcag ctcaaccctg
     1261 gaatgtacca gcaccagaca gtcattgcag accaatttat agtatgcttg cgtcgagatg
     1321 ggcggactgt gtaccagcaa gttctgtcct tggagcttcc aaatgtcctg cgcagctgga
     1381 actggggcct gtgtggttac tttgctttct accacgccct ctatccccga gcctggacgg
     1441 tctatcagct tcctggccag aatgtcaccc tcacctgtcg ccaggtcaca cctatcttgc
     1501 ctcatgacta ccaggacagc agtctccctg taggagtctt tgtgtgggat gtagaaaacg
     1561 aaggagatga gactctggac gtttccatca cgttctccat gcggaatgga ctaggaggcg
     1621 aagatgatgc ggccgggagt ttgtggaatg agcccttccg cctggaacag ggcgggacga
     1681 ctgttcaggg gctcctcttg catcatccaa cccctccgaa cccctacacc atggctgtgg
     1741 ctgcacgatg cacggcagat accacggtaa cccacaccac agcctttgac cccaatggca
     1801 ctggacagca ggtgtggcag gacctacttc aggatggaca gctggactcc cctgctggcc
     1861 aaagcacccc aacacagaaa ggagagggta tcgctggggc tgtatgcgtc tctagcaagc
     1921 tgttacctcg aagccggtgc tgcttggagt tctcactggc ttgggatatg cctaagatca
     1981 tgtttggagc taaaagccaa gtccactaca ggcggtatac acggttcttt ggttcagatg
     2041 gtgatgtggc gcctgccctg agccactatg cactgtgcca ctatgcagac tgggaggaca
     2101 gaatctcagc ctggcagaac ccagtactgg atgacagaac cttgcctgcc tggtacaaat
     2161 ctgcactgtt caatgaactg tacttcctgg ccgatggagg cacagtgtgg ctggaagttc
     2221 ccgcagactc cctaccagag gggctgggag ggagtatgcg tcagcttcgc tctactctgc
     2281 aggactatgg gcgatttggc tatcttgaag gccaggagta ccgcatgtac aacacatacg
     2341 atgtccactt ttatgcgtcc tttgccctcg tcatgctgtg gcccaaactt gagctcagtc
     2401 ttcagtatga tatggctctg gcaactttga aggaggacct gacccggcga cggtacctga
     2461 tgagtggagt ggtggcacct gtgaaaagga ggaacgtcat ccctcatgac attggggatc
     2521 cggatgatga gccatggctc cgggtcaacg catatttgat tcatgatact gctgactgga
     2581 aggacctgaa cctgaagttt gtattgcaaa tttatcggga ctattacctg acgggtgatc
     2641 aaggcttcct ggaggacatg tggcctgtgt gtctggctgt gatggagtcc gaaatgaagt
     2701 ttgacaagga ccaagatgga ctcattgaga atggaggcta cgcagaccag acctatgatg
     2761 catgggtcac cacaggcccc agtgcttact gcggagggct gtggctggcg gcagtggctg
     2821 taatggttca gatggctgtt ctgtgtgggg cccaagatgt ccaggagagg tttgcttcca
     2881 ttctctgccg aggccgagaa gcttatgaga gactgctgtg gaacggacgc tattacaact
     2941 acgacagcag ctcccatcct cagtctcgga gcatcatgtc tgaccagtgt gctgggcagt
     3001 ggttcctgag ggcctgcggc ctgggagaag gagacactga agtatttcct accctgcatg
     3061 tggtccgtgc tctccaaacc atctttgaac tcaacgtcca agcctttgca ggaggagcca
     3121 tgggggccgt gaacgggatg caccctcatg gtgtccctga tagatccagt gtgcagtctg
     3181 atgaagtctg ggtgggtgtg gtctatgggc tggcagccac catgatccaa gagggcctga
     3241 cttgggaagg tttccggaca gctgaaggct gttaccgcac tgtatgggaa cgcctgggcc
     3301 tggctttcca gaccccagag gcatactgcc agcaacaagt gttccgctcc ctggcctaca
     3361 tgcggccact gagcatctgg gccatgcagc tggccctgca acagcagcag cataaaaaga
     3421 gccgcaggcc atcagtcaca caaggcacgg gactaagcac acagcctgaa tgtggaccaa
     3481 agagatcgct ggcaaacctc aattcagagg gatccgacta caaagaccat gacggtgatt
     3541 ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgatatct
     3601 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     3661 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     3721 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     3781 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     3841 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     3901 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     3961 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     4021 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     4081 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     4141 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa
     4201 cgggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     4261 gcaataaaaa gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt
     4321 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     4381 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     4441 agccaacgtc ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg
     4501 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4561 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     4621 tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt
     4681 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     4741 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4801 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     4861 tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca
     4921 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4981 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     5041 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5101 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     5161 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     5221 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     5281 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     5341 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     5401 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     5461 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     5521 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     5581 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     5641 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     5701 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     5761 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     5821 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     5881 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     5941 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     6001 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     6061 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     6121 cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac
     6181 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     6241 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     6301 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     6361 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     6421 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     6481 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     6541 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     6601 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     6661 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     6721 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     6781 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     6841 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     6901 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     6961 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     7021 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     7081 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     7141 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     7201 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     7261 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     7321 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     7381 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     7441 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     7501 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     7561 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     7621 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     7681 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     7741 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     7801 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     7861 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     7921 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     7981 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     8041 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     8101 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     8161 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     8221 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     8281 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     8341 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     8401 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     8461 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     8521 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     8581 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     8641 cgtcgacgga tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct
     8701 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     8761 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     8821 ctgcttagg
//
