LOCUS       V042442                 9582 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042442
VERSION     V042442
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..4262
                     /label="Atp13a2(NM_029097)"
                     /note="Atp13a2(NM_029097)"
                     /gene="Atp13a2"
     CDS             4269..4334
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4358..4946
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5012..5060
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5262..5690
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5704..6033
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6100..6894
                     /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    7070..7191
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7240..7256)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7264..7280
                     /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(7288..7318)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7333..7354
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7642..8230)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8401..9261)
                     /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(9262..9366)
                     /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 ccaccatgag cgcagacagc agcctgctca
      781 tgggcagcac gccccccagc tatgggaccc tgacgacagg gacgtcaatc gatcccctca
      841 gctcctcagc ttcatccgtg aggctcagcg gctactgtgg cagtccatgg agggccatcg
      901 gctatcacgc cgcggtctgg atgctggccg ggatcccttg gctgctattc cgctggaagc
      961 ccttgtgggg cgtgcgcctg cgcctgaagc catgcagcct ggcccacgcc gaaacactcg
     1021 ttatagaaat aaaagacaaa gagggtagct ccagacagct cttcacggtt caggtgcaga
     1081 ctgaggctgt tgttcagggc agcctggagc tgcccccaca ggcccaggca gaggatggcc
     1141 ggagccaggc agccgtggga gtgacaccag agggcacatg gcaggacacc tccgaactcc
     1201 acaggcagga agaggcaaag caggtactgc gctattatgt cctccaaggt caacgctacg
     1261 tctggatgga aacccagcaa gccttctgcc aagtcagcct gctggaccac ggtcgtacct
     1321 gtgatgacgt ccactgctcc agctccggcc tcagcctcca ggaccaagct acgaggaaga
     1381 ccatttacgg ccccaacgtg atcagtattc ctgtcaagtc ctacctgcag ctgctggcgg
     1441 atgaggccct gaatccctac tatgggttcc aagccttcag catcgcactg tggctggccg
     1501 accactacta ctggtatgct ctctgcatct tcctcatctc agccatctcc atctgcctgg
     1561 ctctgtacaa gaccagaaag caaagcttga ccttgaggga catggtcaag ctgtccgtgc
     1621 gggtgcaggt gtgccgaccc ggaggagagg aagagtgggt ggactccagt gagctggtgc
     1681 ccggagactg cttggtgctg ccccaggagg gcggggtgat gccatgcgac gcggctctgg
     1741 tggccgggga gtgtgtggtc aacgagagct ccctgacagg ggagagcacc cctgtgctga
     1801 agacggccct gccagagggg cccaagccct actgcccaga gacccaccgg cggcacaccc
     1861 tcttctgtgg caccctcatt ctgcaggccc gagcctacgt gggaccccgt gtgctagcag
     1921 tggtgactcg gacagggttc tgcacagcca aagggggcct ggtgagctcc atcctgcacc
     1981 caaggcccat cagcttcaag ttctacaaac acagcatgaa gtttgtggct gcgctctccg
     2041 tcctggctct gctaggcacc gtctacagca tcatcatcct ctaccgcaac cgggtacctg
     2101 tgagggagat tgtgatccga gctctggacc tggtgacggt ggtggtgccg cccgccctgc
     2161 ctgccgccat gaccgtgtgc acactctatg cccagagtcg gctgcgcaca cagggcatct
     2221 tctgcatcca cccactgcgc atcaacttgg ggggcaagct gcggctcgtg tgctttgaca
     2281 agacaggcac cctcacggag gacggcttgg acgtaatggg cgtggtgccc ctaaaggggc
     2341 aggtgttgct gccactggtc ccagagccct gccacctgcc cctggggccc ctcctgcgag
     2401 cactggccac ctgtcatgcc ctcagccagc tacatgacac cccggtgggc gaccccatgg
     2461 atctgaagat ggtggagtct acaggctggg tcttagagga gggtccagct gcaggctcag
     2521 cacctgggag ccaggtcttg gtggtaatga gacccccacc cgggggccct cggcagcagg
     2581 aagagccacc agtgccagtc agcgtcctct gccgcttccc cttctcctct gccctgcagc
     2641 ggatggatgt ggtagtgaca tggccagggg ccacccagcc tgaggcctac gtcaaaggct
     2701 ccccagagct tgtggccagc ctctgcagcc ccgagacagt acccagcgac ttttctcagg
     2761 tgctgcagag ctacacagct gctggctacc gagttgtggc cctggctggc aagccactgc
     2821 ccattgcgcc cagccttgcg gccgctcagc agctaacaag ggatactgtg gagcgggaac
     2881 tgagcctcct ggggctgctg gtcatgcgga acctactgaa gccacagacg gccccagtta
     2941 tccagactct gaggaagacg ggcatccgca ccgttatggt gacaggggac aacctgcaga
     3001 cagcagtcac cgtggccaga gcttgtggca tggtgggcgc ccaggagcac ctggctgtca
     3061 tccatgccac ccaccccgag cagggccagc ctgctgccct cgagttcctg ccaacggagt
     3121 cctctgcagt catgaacggg gcgaaggcca caggctatcc cacggtgccg gaaccccagt
     3181 cctgtcacct ggccctcagc gggtccacgt ttgcagtcct tcggaagcac ttccccaagc
     3241 tgctgcccaa ggtcctggtg caggctaccg tctttgcccg aatggccccg gagcagaaga
     3301 cagagctagt gtgcgaactg cagaggcttc agtactgtgt gggcatgtgc ggggatggag
     3361 ccaacgactg tggggccctg aaggcggctg atgtgggcat atcgctgtcc caggcggagg
     3421 cgtcagtggt ctctccgttc acctccagca tggccagtat tgaatgtgtg cccactgtca
     3481 tcagggaagg ccgctgctcc ctggatactt cattcagcgt cttcaagtac atggccctct
     3541 acagcctgac tcagttcatc tctgtgctca ttctctacac aatcaacacc aacctgggag
     3601 acctacagtt cttggccatt gacctggtca tcaccaccac ggttgcggtg ctcatgagcc
     3661 gcacaggccc tgcactgacc ttggtgcgag cgcggccgcc aggggcgctg ctgagtgtgc
     3721 ctgtgcttgg cagtctgctg ctgcaggtgg ccttggtggc cggcatccag ctggggggct
     3781 actttttggt catagctcag ccctggtttg tgcctctgaa cagaactgtg cccgctccgg
     3841 acaacctgcc caactacgag aatacggtgg tcttctctct gtctggcttc cagtacctca
     3901 tcctggccgc agcggtgtct aagggggcac ccttccgcca gccactctac accaacgtgc
     3961 ccttcctggt ggccctggcg ctcttgggct ctgtcctggt gggcctcatc ctggtccccg
     4021 gcctcctgca ggggccccta ggactgagga acattgtgga cagctccttc aagctactgc
     4081 tgctgggtct ggtcgccttc aacttcgtcg gcgcctttat gctggagagc gtgctggacc
     4141 agtgtctccc agcctgcctg cggtggctgc ggcctaaacg ggcctccaag aagcaattca
     4201 agcggctgca acaggagctt gcagaacatc catggcccac tctgcccgtt ggctccgtga
     4261 ggggatccga ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg
     4321 atgacgatga caagtagtga gggcccgata tctcgacaat caacctctgg attacaaaat
     4381 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     4441 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     4501 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     4561 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     4621 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     4681 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     4741 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
     4801 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     4861 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     4921 gatctccctt tgggccgcct ccccgcctgg aaacggggga ggctaactga aacacggaag
     4981 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     5041 gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     5101 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     5161 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     5221 catagcagat ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     5281 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     5341 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     5401 tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga
     5461 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     5521 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     5581 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tggggatttc
     5641 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     5701 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     5761 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     5821 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     5881 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     5941 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     6001 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     6061 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     6121 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     6181 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     6241 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     6301 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     6361 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     6421 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     6481 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     6541 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     6601 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     6661 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     6721 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     6781 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     6841 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     6901 ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     6961 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     7021 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     7081 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     7141 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     7201 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     7261 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     7321 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     7381 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     7441 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     7501 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     7561 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     7621 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     7681 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     7741 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     7801 cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag
     7861 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     7921 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     7981 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     8041 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     8101 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     8161 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     8221 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     8281 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     8341 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     8401 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     8461 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     8521 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     8581 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     8641 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     8701 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     8761 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     8821 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     8881 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     8941 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     9001 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     9061 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     9121 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     9181 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     9241 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     9301 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     9361 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     9421 cccctatggt cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     9481 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     9541 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gg
//
