LOCUS       V035949                 9726 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035949
VERSION     V035949
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..4403
                     /label="Atp13a5(NM_175650)"
                     /note="Atp13a5(NM_175650)"
                     /gene="Atp13a5"
     CDS             4413..4478
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4502..5090
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5156..5204
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5406..5834
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5848..6177
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6244..7038
                     /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    7214..7335
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7384..7400)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7408..7424
                     /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(7432..7462)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7477..7498
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7786..8374)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8545..9405)
                     /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(9406..9510)
                     /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 ccaccatgga gaagagcaag aaggatggcc
      781 accaggctgt gctcaatgaa ggggaggaga atgagctgga ggtgtttggc taccataccc
      841 agaatttacg gagagcactg tgccttgtca cagccatcct gactcttggg gcagttcagc
      901 tgatgttcta ctggaggcct gagtggtggg tgtggaccag ttgcatccca tgccccttgc
      961 aagaagcaga cacaattttg ctgaggacaa ccgatgaatt tcgaagatat atgaggaaaa
     1021 aggtcttctg cctccactta tccacactga agtttcctat aagcaagaac ccagaggaac
     1081 ctctggtagc tgaccaccac tcagtcataa accaagctgt gatgaagcca gaactaaaac
     1141 tacgatgcat ccaagtgcag aaaataaggt acgtgtggga tttcttgaag aagcgatttc
     1201 agaaagttgg gttgctggaa gacagtaact cctgctttga catccaccat acgtttggat
     1261 tgggtctgac caatgaagag caagaggtca gaaggttagt gtgtgggccc aactcgatcg
     1321 aggtggagat ccaacccata tggaagctgc ttgttaaaca ggttttaaat ccattctatg
     1381 tgttccaagc ctttacactc actctgtggc tgtctcaggg ttacatagaa tactctgtgg
     1441 ctatcatcat cttgactgtt atctccattg ttttgagtgt gtatgatttg agacagcaat
     1501 cagttaagct acacaagctt gtggaggagc acaacaaagt ccaggtcacg atcacagtaa
     1561 gagacaaagg tctacaggag ctggagtccc gtctcctggt tccaggagat attctcattc
     1621 ttccagggaa aatttcactg ccctgtgatg ctatcttgat cgatgggagt tgtgtggtga
     1681 atgaaggaat gctcacagga gaaagtatac ctgttacaaa gacaccattg ccccaaacgg
     1741 agaacaccat gccctggaaa tcgcacagtt tggaggacta ccggaaacac gtgctcttct
     1801 gtggaacaga agttattcag gtcaagccat ctgctcaggg gcttgttaga gctgttgttc
     1861 tgcagacagg ttataataca gccaaaggag acttagtgag atccatcctc taccctcgac
     1921 ctctgaactt caaactgtat aatgatgcct tcaagttcat ggtgttcctg gcctgcgttg
     1981 gtgttgtggg atttttctat gccctaggtg tctatatgta ccatgaagta cctccgagag
     2041 aaactgcaac catggccctg atccttctca gtgccaccgt ccctcccgtg ctcccggctg
     2101 cactgaccat aggcaacgtg tacgcccaga agaggttgaa gaaggagaag attttctgta
     2161 tctccccgca gagaatcaac atgtgtggtc agatcaacct cgtgtgcttc gacaaaacag
     2221 gaactctgac agaagatggg ctggacctct gggggactgt ccccaccgct ggcaactgtt
     2281 tccaggcagt ccacagcttt gcctcaggtg aggctgtgcc ctggggccca ctgtgtgcag
     2341 ccatgactag ctgccattct ctgatccttc ttgatgggac catccaagga gaccctttgg
     2401 acctcaaaat gtttgaaggc actggttgga atatggaaga tagccaagtg gcctcctgca
     2461 aatttggcat ggcagattca agcacagtaa taaaaccagg accgaaagcc agccagagtc
     2521 ctgtggacag catcaccatc ttgcgccagt ttccattttc ctcaggcctg caaaggatgt
     2581 ctgtgattgc tcagctggct ggtgaccttc acctccacgt ctacatgaag ggagccccag
     2641 aaatggtggc caggttctgc cgctctgaaa cggtgcctaa gaacttctcg caggagctga
     2701 ggaattacac agtgcaaggc ttccgagtca ttgctcttgc ccacaaaacc ctaaagatgg
     2761 agaggctttc agatatggac catctagcaa gggagaaagt ggagtctgag ttggcctttc
     2821 tgggacttct cataatggag aatcgcttga aaaaggaaac aaggccagtg ctgaaggagc
     2881 tgagtgaggc ccgcatcagg accgtgatgg ttacaggtga caatcttcaa actgccatca
     2941 ctgttgctaa aaattctgaa atgattcctg tgggcagcca ggtcgtcatt gtcgaggcta
     3001 atgagccagg agaccttgtt cctgcttctg tgacctggca gctggtgggg acccaagagc
     3061 caggctctgg gaagaaggac acctacattg acatcggaaa cagctctgta cctgctggga
     3121 aaggttacca ttttgcaatg agtgggaagt cataccaagt gttatttcat catttctaca
     3181 gcatgctgcc acaaattctg gttaatggaa ccatttttgc aagaatgtct cctgggcaga
     3241 agtcaagcct cgtggaagag tttcagaaat taaattatta tgtgggcatg tgtggagatg
     3301 gagccaatga ctgtggggct ttgaaaatgg ctcatgctgg catctcacta tcagaacagg
     3361 aagcatccgt ggcatctccc ttcacctcga aaacagccaa tatcgagtgt gtgcctcatc
     3421 tcatcaggga gggccgggct gctctggtct catccttcgg tgtatttaaa tacctgacta
     3481 tgtacggcat catccagttt attggaacat cacttctgta ctggcaacta cagctctttg
     3541 gtaactacca gtacctcctg caagatgtgg ccatcacgct gatggtcagc ttaacaatga
     3601 gcataaacca tgcctacccg aagttggctc cctacagacc agcaggacaa ctactttctc
     3661 ctcagttgct actctcagtc ttcatgaatt cctgttttac ctgcattgtg caagtgtgta
     3721 catttctcac cgtgaaacag cagccttggt actgcgaggt ctacaaatac agtgaatgct
     3781 ttctggtcaa ccaaagtaac ctctcagcca acgtgagttt ggaccgaaac tggactggaa
     3841 atgcaactct ggttcccgct tctgtgctaa gtttcgaggg cactacgttg tggcccatcg
     3901 tcaccttcaa ctgcatctct gcagcattta ttttttctaa gggaaagccg tttcggaaac
     3961 ccatctacgc aaactattta ttctcccttc tgctggcatc tgctgcgggc ctcactattt
     4021 tcattctgtt ttgtgatttt caagacctgt accgtaaaat ggagttcatc ccaaccccca
     4081 catcatggag agtttcaatt ctgatagcag cttttgtcca gttctgcgtt gccttctttg
     4141 tagaggatgc cgtccttcaa aacagagagc tctggctgtt tatcaagaaa gaatttggat
     4201 tctactccaa aagtcagtat aggatcttgc agagaaagct ggcagaggac tctacatggc
     4261 ctcccgtgaa caggacagat tatgcagtca atggcaaaaa tggattctat gtcaacagag
     4321 cctatgagag ccccgaagag gtccccaaag ggaagctcaa gctggaagag caggcttcag
     4381 aacagcactt ttggactaga ctgtaaggat ccgactacaa agaccatgac ggtgattata
     4441 aagatcatga catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga
     4501 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     4561 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     4621 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     4681 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     4741 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     4801 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     4861 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     4921 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     4981 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     5041 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg
     5101 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     5161 ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg
     5221 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     5281 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     5341 caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta
     5401 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     5461 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     5521 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg
     5581 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     5641 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     5701 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     5761 tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     5821 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     5881 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     5941 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     6001 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     6061 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     6121 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     6181 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     6241 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     6301 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     6361 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     6421 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     6481 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     6541 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     6601 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     6661 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     6721 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     6781 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     6841 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     6901 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     6961 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     7021 cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc
     7081 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     7141 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     7201 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     7261 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     7321 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     7381 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     7441 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     7501 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     7561 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     7621 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     7681 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     7741 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     7801 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     7861 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     7921 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     7981 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     8041 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     8101 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     8161 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     8221 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     8281 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     8341 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     8401 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     8461 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     8521 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     8581 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     8641 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     8701 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     8761 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     8821 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     8881 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     8941 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     9001 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     9061 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     9121 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     9181 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     9241 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     9301 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     9361 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     9421 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     9481 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     9541 cgacggatcg ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat
     9601 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     9661 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     9721 cttagg
//
