LOCUS       V023355                 8346 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023355
VERSION     V023355
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..3026
                     /label="Sun1(NM_001007147)"
                     /note="Sun1(NM_001007147)"
                     /gene="Sun1"
     CDS             3033..3098
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3122..3710
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3776..3824
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4026..4454
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4468..4797
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4864..5658
                     /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    5834..5955
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6004..6020)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6028..6044
                     /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(6052..6082)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6097..6118
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6406..6994)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7165..8025)
                     /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(8026..8130)
                     /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 ctcggatccg ccaccatgga cttttctcgg ctgcacacat
      781 acaccccacc ccagtgtgtg ccagagaaca ctggctatac ctacgcactc agttctagtt
      841 attcttcgga tgctctggat tttgagactg aacacagatt ggaacctgtc tttgactctc
      901 caaggatgtc ccgccgtagc ttgcgcctga tcacaacaac agctgcatac agcagtgggg
      961 acggccagac tgttgacact cacatcagca ccagcagggc caccccggcc aaggggagag
     1021 aaaccaggac agtcaaacag agaagtgcaa gcaagccagc ttttagtatc aaccacctgt
     1081 cagggaaggg cttgtcctca agcacaagcc atgacagctc ttgcagcctg cggagtgcca
     1141 cggtgctgcg gcatcctgtg ctagacgagt ctctgattcg tgagcagacc aaagtggacc
     1201 acttctgggg tctcgatgat gatggcgacc ttaaaggtgg aaataaagct gccactcagg
     1261 gaaatggtga actggcagca gaggtggcga gcagcaacgg atacacctgc cgcgactgca
     1321 ggatgctctc ggctcggaca gacgctctca cagcccactc tgccgtccac gggcccacct
     1381 caagggttta ctccagagat aggactctca gaccccggaa ggcagcctct ggaaccttct
     1441 ggtggctagg gagtggctgg taccaatttg ttactttgat ttcctggctg aatgtgtttc
     1501 ttcttaccag gtgccttcga aatatttgca aggtttttgt cttgctgctc ccactcctac
     1561 ttttattagg tgctggtttc tccttgtggg gccagggcaa cttcttctca ctcctaccaa
     1621 tgttgaattg gacggccatg cagccagcgc agagggtgga caatcccaag gacatgcaca
     1681 gacccggccc tctttccccg agtccacctc tcaaggttga tcccatggct tcccagtggc
     1741 ctcaggagag tgatatgggg cagaagatag cctctttgag tgcacagtgc cacaaccatg
     1801 atgagagact cgcagagctg acagtcctgc ttcagaaact ccagatacgg gtagaccaag
     1861 tggatgacgg cagggaaggg ctgtcactgt gggtcaagga tatggttgga cagcacctgc
     1921 aggagattgg ctccatagaa ccacctgatg ctaagactga cttcctgact ttacaccatg
     1981 accatgaagt gcgtctctcc agtttggaag atgttcttag aaaactgact gaaaaatctg
     2041 aggctatcca gaaggagctg gaggaaacca agctgagagc aggcagcagg gatgaagagc
     2101 agcccctcct tgaccgtgtg cagcacctag aactggagct gaacctgctg aagtcacagc
     2161 tgtcggactg gcagcatctg aggagcagct gtgagcaggc tgatgcccgc atccaggaga
     2221 ctgtgcagct catgttctct gaggaccagc cgggcggctc cctcgagtgg ctgttacaga
     2281 agctttcttc tcggttcgtg agcaaggatg agctgcaggt gcttttgcat gaccttgagc
     2341 tgaaactact gcagaatatc acacaccaca tcaccgtcac aggccaggct ccgacatccg
     2401 aggccattgt gtctgccatg agtcaggcag ggatttcagg aatcacagaa gcgcaagcac
     2461 atatcattgt gaacaatgct ctgaggctgt actcccaaga caagactggg atggtggact
     2521 ttgctctgga gtctggaggt ggcagcatcc taagcactcg gtgctctgag acctatgaga
     2581 ccaagacggc actgctgagc ctgttcgggg tcccactgtg gtacttctca cagtcacctc
     2641 gagtggtaat ccagcccgac atctacccgg ggaattgctg ggcatttaag ggttcccagg
     2701 ggtacctggt agttcgcttg tccatgaaga tctacccaac cacgttcacc atggagcaca
     2761 ttccaaagac gctgtcaccg actggtaaca tctccagtgc ccccagagac tttgcagtct
     2821 atggactgga aaccgagtat caggaagagg ggcagcctct gggacggttc acctatgacc
     2881 aggaagggga ctcactccag atgttccaca cactggaaag acctgaccaa gggttccaga
     2941 tagtagagct ccgggttctg tccaactggg gccaccctga gtacacttgc ctctaccggt
     3001 tccgagtcca cggagagccc atccagctcg aggactacaa agaccatgac ggtgattata
     3061 aagatcatga catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga
     3121 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     3181 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     3241 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     3301 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     3361 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     3421 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     3481 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     3541 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     3601 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     3661 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg
     3721 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     3781 ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg
     3841 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     3901 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     3961 caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta
     4021 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     4081 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     4141 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg
     4201 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     4261 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4321 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     4381 tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     4441 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4501 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     4561 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4621 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4681 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     4741 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     4801 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     4861 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4921 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4981 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     5041 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     5101 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     5161 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     5221 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     5281 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     5341 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     5401 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     5461 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     5521 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     5581 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     5641 cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc
     5701 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     5761 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5821 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5881 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5941 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     6001 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     6061 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     6121 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     6181 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     6241 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     6301 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     6361 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     6421 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     6481 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     6541 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     6601 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     6661 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6721 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6781 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6841 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6901 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     6961 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     7021 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     7081 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     7141 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     7201 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     7261 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     7321 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     7381 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     7441 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     7501 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     7561 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     7621 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     7681 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     7741 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     7801 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     7861 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7921 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7981 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     8041 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     8101 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     8161 cgacggatcg ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat
     8221 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     8281 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     8341 cttagg
//
