LOCUS       V025928                 9085 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025928
VERSION     V025928
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"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             922..987
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             991..4605
                     /label="ATP2B4(NM_001684)"
                     /note="ATP2B4(NM_001684)"
                     /gene="ATP2B4"
     polyA_signal    4685..4909
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4955..5383
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5397..5726
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5793..6587
                     /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    6761..6882
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6931..6947)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6955..6971
                     /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(6979..7009)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7024..7045
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7333..7918)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8089..8949)
                     /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(8950..9054)
                     /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 aagccaccat ggactacaaa gaccatgacg gtgattataa agatcatgac
      961 atcgactaca aggatgacga tgacaagctt atgacgaacc catcagaccg tgtcttgcct
     1021 gccaactcga tggccgagag ccgtgaaggg gactttggct gcacagtaat ggaactgagg
     1081 aagctcatgg agctgcgttc aagggatgca ctgacccaga ttaatgtcca ctatggaggt
     1141 gtacagaatc tctgcagtag actgaaaacc tcccctgtgg aaggtctgtc tgggaaccct
     1201 gcagatctgg agaaacgtag gcaggtgttt ggacacaacg tgatcccccc caaaaagccc
     1261 aagactttct tagaattagt gtgggaagct cttcaagatg tcacgcttat catcctggag
     1321 attgcagcca tcatctccct ggtcctgtcc ttttatcgcc ctgctggtga agaaaatgaa
     1381 ctgtgtggtc aagtcgcaac taccccagaa gatgaaaatg aggcacaagc tggctggatt
     1441 gagggggcag ccatcctttt ctcagtgatc atcgtggtgt tagtgactgc ctttaatgat
     1501 tggagcaaag agaagcaatt ccgggggctg cagtgccgca ttgaacagga gcaaaagttc
     1561 tccatcatcc gaaacggtca actcatccag ctccctgtgg ctgagattgt ggttggtgat
     1621 attgcccaag tcaaatacgg tgatctgctg cctgcagatg gaatcctgat ccaagggaat
     1681 gatctgaaga ttgatgagag ctctctgaca ggggaatctg accatgtcaa gaagtccctg
     1741 gacaaagacc ccatgttgct ctcagggacc catgtcatgg aaggttctgg ccggatggtg
     1801 gtgacagctg ttggtgtcaa ctctcagact ggaatcatcc ttactctctt gggggtcaat
     1861 gaggatgacg aaggggagaa aaagaagaaa ggtaaaaaac aaggagtccc tgaaaatcgc
     1921 aacaaagcaa agacccaaga cggagtggcc ctggaaatcc agccactcaa cagccaggag
     1981 ggaatcgaca atgaggaaaa ggacaagaag gcagtcaagg tgcctaaaaa ggagaagtca
     2041 gtgctgcagg gcaagctgac tcgcctggct gttcagattg ggaaagccgg tctgctcatg
     2101 tctgctctca cggttttcat cctgattcta tactttgtga ttgacaactt tgtgataaat
     2161 cgcagaccat ggctccctga gtgtactccc atctacatcc agtactttgt caagttcttc
     2221 atcatcggca tcactgtact ggtggtggct gtgccagagg ggctgcctct ggctgtcacc
     2281 atctcactgg cctactctgt gaagaaaatg atgaaagaca ataacctagt acggcacttg
     2341 gatgcttgtg agaccatggg caacgccacc gccatctgct ctgataagac aggcacgttg
     2401 accatgaacc gcatgactgt ggtacaagct tatattgggg gcatccatta ccgtcaaatc
     2461 ccaagccctg atgtcttcct gcccaaagtc ctggacctca ttgtcaatgg catttctatc
     2521 aacagtgctt atacctccaa gattctgcct ccagagaagg agggaggcct gcctcggcag
     2581 gtgggcaaca agaccgagtg tgctctgcta ggctttgtca cagatctgaa gcaggattat
     2641 caggctgtgc gtaatgaagt gcccgaggag aagctctaca aggtgtacac ctttaactca
     2701 gtgcgcaagt caatgagcac cgtcatcagg aatcccaacg gtggcttccg tatgtacagc
     2761 aagggcgcct ctgagatcat cttgcgcaag tgtaatcgaa tcctggaccg gaaaggggaa
     2821 gcagtgccat tcaagaataa agacagagat gatatggtac gcactgtcat cgagcccatg
     2881 gcctgtgatg gactccggac tatctgcata gcttaccggg acttcgatga cacagagccc
     2941 tcttgggaca atgagaatga gatcctcacc gaactgacct gtatcgcggt ggtgggcatt
     3001 gaggaccctg tgcgcccaga ggtgccagat gctattgcca aatgcaaaca agctggcatt
     3061 actgtcagaa tggtgacagg tgacaacatc aacacagccc gggccattgc caccaaatgt
     3121 ggcattctga cacctgggga tgacttcctg tgcttagaag gcaaagaatt caaccggctc
     3181 atccgcaacg agaaaggcga ggtagagcaa gaaaagctgg acaagatctg gcctaagctt
     3241 cgggtcctgg cgcgatcttc tcccactgac aagcacaccc tggtgaaagg cataattgac
     3301 agcactgttg gggaacaccg gcaggtcgtg gctgtcactg gtgatggcac aaatgacggg
     3361 cctgctctga agaaagcgga tgttggtttt gccatgggca tcgcaggcac agatgtagca
     3421 aaggaggctt cagacatcat cctaacagat gacaacttca ccagcattgt gaaggcagtg
     3481 atgtggggac gaaatgtcta tgacagcatc tccaagttcc tgcagttcca gctcactgtc
     3541 aatgtggtgg ccgtgattgt agccttcact ggagcctgta tcactcagga ttccccattg
     3601 aaagctgtgc agatgttgtg ggttaatctg atcatggaca cttttgcttc attggccctg
     3661 gccacagagc cccctacgga atctctgttg aagcggcgcc cctatggccg aaataagcct
     3721 ctgatctcac gcactatgat gaagaacatc ttgggccatg cattctatca gctcattgtc
     3781 atctttatcc ttgtctttgc gggtgagaaa ttctttgata ttgatagtgg gaggaaggca
     3841 cctctacatt caccacccag ccagcactat accattgttt ttaacacctt cgtgctgatg
     3901 cagctcttca atgaaatcaa ctcccgaaag atccatggag agaagaacgt cttttcaggc
     3961 atctaccgca acattatctt ctgctctgta gtcttgggca cattcatctg ccagattttc
     4021 atcgtggaat ttgggggtaa acccttcagt tgtacaagcc tcagcctgtc tcagtggctg
     4081 tggtgtctct tcattgggat tggagaactt ctgtggggcc agttcatctc cgcaatacct
     4141 acccgatccc tgaagttcct gaaggaggct gggcatggca ccaccaaaga ggagatcacc
     4201 aaggatgccg agggactgga tgagattgac catgctgaga tggagctgcg ccgaggccag
     4261 atcctctggt tccggggcct gaaccgtatc cagactcaga tcaaagtggt caaagcgttc
     4321 catagttccc tccacgaaag cattcagaaa ccctacaacc aaaagtccat ccacagcttc
     4381 atgacccacc ctgaattcgc catagaggag gagttgccac gaacaccact cctggatgag
     4441 gaagaggagg aaaatcctga caaggcttct aagtttggga ctagggtgct cctgttggat
     4501 ggtgaggtca ctccatatgc caatacaaac aacaatgcgg tggattgcaa ccaagtgcag
     4561 ctcccccagt cggacagctc tctacagagc ctagagacat cagtttgaga attctgcaga
     4621 tatccagcac agtggcggcc gctcgagtct agagggcccg tttaaacccg ctgatcagcc
     4681 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     4741 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     4801 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     4861 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     4921 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     4981 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     5041 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     5101 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     5161 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     5221 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     5281 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     5341 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     5401 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5461 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     5521 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     5581 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     5641 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     5701 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     5761 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     5821 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     5881 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     5941 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     6001 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     6061 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     6121 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     6181 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     6241 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     6301 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     6361 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     6421 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     6481 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     6541 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     6601 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     6661 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     6721 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     6781 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     6841 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     6901 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     6961 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     7021 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     7081 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     7141 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     7201 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     7261 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     7321 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     7381 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     7441 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     7501 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     7561 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     7621 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     7681 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     7741 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     7801 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     7861 tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     7921 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     7981 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     8041 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     8101 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     8161 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     8221 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     8281 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     8341 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     8401 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     8461 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     8521 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     8581 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     8641 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     8701 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     8761 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     8821 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     8881 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     8941 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     9001 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     9061 tccccgaaaa gtgccacctg acgtc
//
