LOCUS       V046041                 9570 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046041
VERSION     V046041
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..4250
                     /label="Spag5(NM_017407)"
                     /note="Spag5(NM_017407)"
                     /gene="Spag5"
     CDS             4257..4322
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4346..4934
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5000..5048
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5250..5678
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5692..6021
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6088..6882
                     /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    7058..7179
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7228..7244)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7252..7268
                     /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(7276..7306)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7321..7342
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7630..8218)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8389..9249)
                     /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(9250..9354)
                     /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 ccaccatgtg gagggtgaaa acactgaatc
      781 tcggtttgtc gccttcgccc caaaagggga agccagctat gagtactcct ctccgagagc
      841 ttaaactaca gcccgaggcc ctcgccgatt cagggaaagg tccctctatg atctccgcgc
      901 tgaccccata cctgtgcagg ctggagctga aggaaagatg caacaactca tctccagtgg
      961 attttatcaa tactgagaac aacttccttt cagaacagtt cagccatcct tcaacgcaca
     1021 tagaagcttg ccagcgtgaa tccgatccaa ctcctgaaag caactctctt ttccatactt
     1081 tggaggaagc aatagaaaca gtggatgact ttgtcgtgga cccaagagat gatagtatag
     1141 tggagtccat ggttcttctg cccttttcat tagggcagca gcaagacctg atgctccagg
     1201 ctcacttaga taccacagca gagagaacta aaagctctct aaatgaatct ttggggctag
     1261 aagatctggt ggggaaggag gtggcacctt gtgtggaaga cagcctaaca gaaattgttg
     1321 ctattaggcc cgagcaacct acatttcagg accctccgtt aggacccagt gacactgaag
     1381 atgcacctgt ggacttagtt ccttctgaaa atgtcctgaa tttctctctg gctcgtcttt
     1441 ctccttcagc tgtcctggcc caggatttct ctgtcgatca tgtcgatcca ggggaggaaa
     1501 ccgtagagaa cagagtccta caggaaatgg aaacgagctt ccccacattc cctgaggagg
     1561 ccgaattggg agaccaagca cctgctgcaa atgcagaagc cgtctcaccc ttgtacctga
     1621 catcaagtct agtggagatg gggccccggg aagccccagg cccaacagta gaagacgcca
     1681 gtaggattcc tggccttgag tcagagactt ggatgtcccc attggcctgg ctagaaaagg
     1741 gtgtgaatac atcggtcatg ctacaaaacc tccgccagag cttatccttt tcttctgtgc
     1801 ttcaggatgc tgccgttggc aacacacccc tcgccacgtg ttctgtgggc acttccttta
     1861 ctcctccagc accactggag gtaggcacca aggacagtac ttcagagaca gagcgcctcc
     1921 tcttgggctg ccgtcctcca gatctggcta ccttgtcccg acacgacttg gaagagaacc
     1981 tgttgaactc tcttgtcttg ctggaagttc tttcccacca gctacaagcc tggaagagcc
     2041 agttgactgt ccctcaccgg gaagctcgag acagtagtac acagactgac agctctcctt
     2101 gtggggtcac taagacacct aaacatcttc aggacagcaa ggagattaga caggctctac
     2161 tgcaagccag gaatgtcatg caatcatggg gtctagtctc tggagacctg ttgtccttgc
     2221 ttcacctgtc cctaacacac gtgcaagaag gcagagtgac agtgagtcag gagtctcagc
     2281 ggtcaaaaac cctggtttcc tcctgttctc gtgtgctgaa gaaattgaag gcaaagctgc
     2341 agagcctcaa aacagaatgt gaggaggcaa ggcacagcaa ggaaatggcc cttaaaggca
     2401 aggctgcggc agaggcagta ctggaggctt tccgtgcaca tgccagccag cgcatcagcc
     2461 aactggaaca gggtctcaca tctatgcagg aattcagagg ccttctacag gaggcacaga
     2521 cccaactgat agggcttcac acagaacaaa aggagctggc tcagcagaca gtgagtctga
     2581 gttctgcctt gcagcaggac tggacatcag tccaattgaa ttatggaata tgggcagctt
     2641 tgctgagttg gtctcgagaa ctcaccaaga aactcacagc caagagccgg caggcccttc
     2701 aggaacgtga tgctgcaatt gaagaaaaga agcaggttgt gaaggaagtg gaacaagtct
     2761 ctgcccattt agaggactgt aaaggtcaaa tcgaacaact gaagttggaa aatagtcgcc
     2821 ttactgcaga tctctcggct cagctgcaga ttctgactag cacagagagt cagctaaagg
     2881 aagtacggag tcagcattcc cgctgtgtcc aggacctggc cgtgaaggat gaattgctgt
     2941 gtcaacttac ccagagcaac aaggagcagg ctactcaatg gcaaaaggaa gaaatggaac
     3001 tgaaacacat acaagcagaa ctgctacagc agcaggctgt gttggctaag gaggtccagg
     3061 acctgaggga gactgtggag tttatagatg aagaaagtca agttgctcac cgggagctgg
     3121 gccagattga gagtcagttg aaagtcacgc tagagttact gcgggagcgc agcctgcagt
     3181 gtgagaccct cagggacact gtggacagcc tgagggctga gctggccagc actgaagcaa
     3241 agcatgagaa acaggcccta gagaagacac accagcattc tcaagagctg cggctactgg
     3301 ctgagcagct gcagagtctc accctcttct tacaggcaaa actcaaggag aacaaggctg
     3361 aatcagagat cattctgccc agcacaggct ctgctccagc ccaggaacac cctctgtcca
     3421 atgacagcag catctcagaa cagaccccga cagcagcagt agatgaagtg ccagaaccag
     3481 ctcctgtgcc attgcttgga agtgttaaga gtgctttcac ccgagtagcc tcaatggctt
     3541 cctttcagcc tacagagacc ccagacttgg agaagagcct ggcagaaatg agtactgtgt
     3601 tacaagagct taagagcctg tgttccctgc tgcaagagtc taaagaggag gccactgggg
     3661 tcctgcagag ggaaatctgt gaactacact cgagactaca ggcccaagaa gaagagcatc
     3721 aggaagccct gaaggcaaag gaagcagaca tggagaagct gaaccaggcc ttgtgcttgc
     3781 tccgcaagaa tgagaaggag ctcctggaag tgatacagaa gcagaacgag aagatcctgg
     3841 ggcaaataga caagagcggc cagctcataa acctcagaga ggaggtgacc cagctcacac
     3901 agtcacttcg gcgtgcagag acagagacta aagtgctcca ggaagccctg gaaggccagc
     3961 tagatcccag ctgccagctg atggctacta actggatcca ggaaaaagtg tttctctcac
     4021 aggaggtgag caagctgagg gttatgttcc tggagatgaa aactgagaag gaacagctga
     4081 tggacaagta tctgagccat aggcacatcc tggaggagaa tcttcggcgc tctgacacag
     4141 agttaaagaa actcgatgac acaattcagc atgtctatga gactctgttg tctatcccag
     4201 agactatgaa gagttgcaag gagttacaag gattgctaga atttctgagc ggatccgact
     4261 acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca
     4321 agtagtgagg gcccgatatc tcgacaatca acctctggat tacaaaattt gtgaaagatt
     4381 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     4441 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     4501 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     4561 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     4621 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     4681 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     4741 gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     4801 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     4861 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     4921 ggccgcctcc ccgcctggaa acgggggagg ctaactgaaa cacggaagga gacaataccg
     4981 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg gtgttgggtc
     5041 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     5101 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     5161 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcagatct
     5221 gcgcagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     5281 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     5341 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     5401 tcggggcatc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     5461 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     5521 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     5581 ccctatctcg gtctattctt ttgatttata agggattttg gggatttcgg cctattggtt
     5641 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     5701 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     5761 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     5821 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     5881 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     5941 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     6001 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     6061 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     6121 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     6181 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     6241 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     6301 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     6361 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     6421 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     6481 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     6541 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     6601 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     6661 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     6721 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     6781 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     6841 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     6901 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     6961 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     7021 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     7081 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     7141 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     7201 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     7261 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     7321 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     7381 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     7441 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     7501 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     7561 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     7621 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     7681 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     7741 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     7801 ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt
     7861 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     7921 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     7981 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     8041 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     8101 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     8161 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     8221 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     8281 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     8341 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     8401 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     8461 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     8521 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     8581 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     8641 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     8701 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     8761 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     8821 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     8881 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     8941 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     9001 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     9061 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     9121 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     9181 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     9241 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     9301 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     9361 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtcg
     9421 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     9481 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     9541 accgacaatt gcatgaagaa tctgcttagg
//
