LOCUS       V045546                10059 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045546
VERSION     V045546
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..4739
                     /label="Clspn(NM_001405798)"
                     /note="Clspn(NM_001405798)"
                     /gene="Clspn"
     CDS             4746..4811
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4835..5423
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5489..5537
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5739..6167
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6181..6510
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6577..7371
                     /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    7547..7668
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7717..7733)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7741..7757
                     /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(7765..7795)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7810..7831
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8119..8707)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8878..9738)
                     /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(9739..9843)
                     /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 ccaccatgac cggcgaggtg ggttctgagg
      781 ttaacctgga agtcaacgat ctaaaacttc tttcacaaga agcagcagac agtcctgtag
      841 acagtggaca gggcagcttt gaaacactcg agcccttgag tgaaagagat tctgatgaag
      901 agatttttgt gagcaagaaa cctaagagca ggaaagtgtt acaagacagc gattctgagg
      961 ctgaggacag agacgatgct ccggagaaac ccacctacga cgacagcgca gaggacactc
     1021 aggagaactt acattcaggg aagagtcaga gcagaagctt tcccaaagcc ctggctgaca
     1081 gtgatgagag tgacatggag gaaactccga gtcaggagag tcctgagact caagaggctc
     1141 cttccttgga gccaggccac cagacaggac actctgtgga cttcaccacc ggcagaaagc
     1201 tctccaaaac actcctcaga gaaggagctg agggaaaagc aaaatccaag agacggctgg
     1261 agaaagagga gagaacaatg gagaaaatca ggagactaaa aaagaaggaa acaagatgtg
     1321 aggaaagtga tgcagaccgg ccgcttaatg acagtggctg tctccttgaa gatagtgatc
     1381 tttttgagac aggtttggag gaagaaaatg actctgcact ggaggatgaa gagtcgttag
     1441 agtcaataag ggcggctgta aaaaacaagg tgaaaaatcg caagaaaaag gaaccaactc
     1501 tggagagtga ggctttttca ttagaagacg gaaatgagtt atccaaaggc agtgcgagga
     1561 aggaaagaaa ggcagcgagg ctaagtaagg aagcattaaa aaaactgcat agtgagaccc
     1621 agcgtctggt ccgagaatct gcacttaatc ttccatatca tatgcctgag agtaaaacta
     1681 ttcacgattt cttcaaacgt aagccccggc ccacgtgcca gggcagtgcc atggccctgc
     1741 tgaagtcatg taagtatcag tcgggccatt acaaggaaac cgtaaaccct gcagatgcag
     1801 ctggaatggg tgctgaggac tccagcagag gttctgagca gaggacaggg gctggaattg
     1861 cagcggaaac taatgttctc tctgaggtct cagaagaagc cgggatcact gctggatcag
     1921 atgaggcttg tgggaaggat ccggtaagac gaggagagct ggaaattgag gagactgaga
     1981 agcacagtga tgacagacct tattctcctg gggacagatc catgtcacag caggaatcct
     2041 ccatccccag gatcgaagac aatgaggggc atcaggctgg agacctcacc gaatctgacc
     2101 ctcctgccct ggagggagaa gaactgaaaa cagtagaaaa aacagacgca aaagagggaa
     2161 tgcctgaaca gaaaacccag tctgcagcag cagcggcagt ggcagtagtg acggcggcgg
     2221 cggcgccacc tgaaaaggtg agacgattca ccgtggaccg acttagacag ctgggagtgg
     2281 atgtttccag tcaacctcgg ctgggcgctg atgaagattc ctttgtgata cttgatgaac
     2341 ctaaaaccaa cagagaattg gaagccttga agcaacgttt ctggaggcac gctaacccag
     2401 cagccagtcc cagggcttgt cagacggtga acgtgaacat catcgtgaag gacttgggca
     2461 ctaacgggaa ggaggagctg aaggcagaag tggtacccgt cactttggca gcagagaagc
     2521 tggagggagc aagccacgca aagccaggtg aaaaactgca gatgttgaaa gctaagttgc
     2581 aggaggcaat gaaactccga aggcttgagg agcggcagaa gcgccaagca ttatttaaat
     2641 tagataatga agatggattt gaagaggagg aggaagagga agaaatgaca gatgagtctg
     2701 aggaagatgg agaagaggag gtgggagagg aggaggagga ggatcaagac attcaggaga
     2761 ctacagaata tcttcttggt agtgaagata cagaaaccaa agatgaaaaa gaaacagata
     2821 aagaaaatac agacaccagt agtgacattg gcaagtctgt tgccctctgt gtccccaagc
     2881 ctctctcttc agactccacc ttacttctgt ttaaggacag ctcttctaag atgggttact
     2941 ttcctactga agaaaagtca gaaacagatg aatacttggc caagcagtct gacaaactgg
     3001 atgaggatga ctcgagttca ctgctaacaa aggagagcag tcacaatagc agctttgagc
     3061 tgattggttc cacaattcca tcctatcagc cttgcaacag acagataggc cgtggggcaa
     3121 gttttctccc tacagcagga ttcagatcgc cgtcccctgg gctgttccga gggagtttga
     3181 tcagctcggc ttctaagagt tcaggaaaac tgtctgagcc ttcacttccc gtagaggatt
     3241 cccaggatct gtacaccgcc tccccagagc ctaagacact tttcctagga gcaggagact
     3301 tccagttctg tttggaagat gacactcaga gccaactgtt ggatgcagat ggatttttaa
     3361 acattagaaa tcaccgccat cgatatcagg ctgtgaagcc ccagttgcca ctggccagca
     3421 tggacgagaa tgccatggac gccaacatgg acgagctgct ggatttgtgc actggacagt
     3481 ttacatctca gcctgaggag aaatgtcaac caaggaagaa tgacaaaaaa gaaaacatgg
     3541 aggaactttt gaatctctgc tcgggcaaat ttcctactca agatgcctcc ccggtggccc
     3601 cactggggct gaggagtcag gagaaggaga gcagcacaga agatccgatg gaagaggcac
     3661 ttgctctttg ctccggctct ttcccaacag acagggagga agagggtgaa gaggaggaat
     3721 ttggagactt ccagcttgtt tcaaaggaga atggatttgc tagtgacgag gatgaacaca
     3781 gtgactccaa tgatgaagag ctggccctgg acctggagga tgatgaagaa gagctcctga
     3841 agcagtctga gaagatgaag cgccaaatga gactgaagaa atacttggaa gatgaggcag
     3901 aggtgtcagg cagtgacgtg ggaagtgagg acgagtacga tggggaggag attgatgaat
     3961 acgaagagga cgtcattgat gaggtgcttc cgtctgatga ggagctggag agtcagatca
     4021 agaagataca catgaaaacc atgttggatg atgataagcg gcgactacgg ttatatcaag
     4081 agaggtacct tgctgatggg gatctacaca gcgacgggcc tggacgaaca agaaaattcc
     4141 gatggaaaca catagatgat acctcccaga tggatctatt tcacagagac tccgatgacg
     4201 atcaggttga agaacagctt gacgagaccg aagccaagtg gaggaaggaa cggattgagc
     4261 gagagcagtg gcttcgggaa caggctcaac aggggaaaat tgctgctgac gaagaggaca
     4321 ttggggacga cagtcagttt atgatgctag ccaagaaagt cacagccaaa gcactgcaga
     4381 agaatgccag tcatactgtg gttgttcagg aatcaaagtc tgtgctcaga aacccttttg
     4441 aaaccatcag accaggaggt gctcaccagc tgaagacagg ctccctgcta aaccagccca
     4501 aagctgtgct ccaaaaactt gctgccctgt ctgacctcaa ccccagcgct ccccgaaact
     4561 caaggaactt tgtctttcat acactttctc ccaccaaggc agaggcagca aaggactcat
     4621 ctaagcctca ggtcagaaga aggggactgt cgtcaatgat gtcaccgtct cctaagcggc
     4681 taaagacaaa cggaagcagt ccagggccaa agcgaagcat cttcagatac ttggaaagcg
     4741 gatccgacta caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg
     4801 acgatgacaa gtagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg
     4861 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     4921 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     4981 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     5041 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     5101 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     5161 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     5221 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     5281 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     5341 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     5401 ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag
     5461 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg
     5521 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     5581 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     5641 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     5701 agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     5761 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     5821 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     5881 agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     5941 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     6001 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     6061 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc
     6121 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     6181 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     6241 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     6301 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     6361 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     6421 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     6481 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     6541 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     6601 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     6661 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     6721 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     6781 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     6841 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     6901 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     6961 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     7021 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     7081 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     7141 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     7201 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     7261 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     7321 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     7381 tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     7441 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     7501 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     7561 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     7621 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     7681 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     7741 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     7801 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     7861 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     7921 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     7981 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     8041 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     8101 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     8161 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     8221 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     8281 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     8341 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     8401 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     8461 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     8521 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     8581 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     8641 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     8701 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     8761 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     8821 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     8881 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     8941 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     9001 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     9061 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     9121 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     9181 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     9241 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     9301 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     9361 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     9421 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     9481 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     9541 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     9601 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     9661 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     9721 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     9781 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     9841 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     9901 ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     9961 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
    10021 caaggcttga ccgacaattg catgaagaat ctgcttagg
//
