LOCUS       V035407                 8316 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035407
VERSION     V035407
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..2996
                     /label="RAD54L(NM_003579)"
                     /note="RAD54L(NM_003579)"
                     /gene="RAD54L"
     CDS             3003..3068
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3092..3680
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3746..3794
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3996..4424
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4438..4767
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4834..5628
                     /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    5804..5925
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5974..5990)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5998..6014
                     /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(6022..6052)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6067..6088
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6376..6964)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7135..7995)
                     /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(7996..8100)
                     /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 gaggagcttg gctcccagcc
      781 agctggccaa gagaaaacct gaaggcaggt cctgtgatga tgaagactgg caacctggcc
      841 tagtgactcc taggaaacgg aaatccagca gtgagaccca gatccaggag tgtttcctgt
      901 ctccttttcg gaaacctttg agtcagctaa ccaatcaacc accttgtctg gacagcagtc
      961 agcatgaagc atttattcga agcattttgt caaagccttt caaagtcccc attccaaatt
     1021 atcaaggtcc tctgggctct cgagcattgg gcctgaaaag ggctggggtc cgccgggccc
     1081 tccatgaccc cctggaaaaa gatgccttgg ttctgtatga gcctcccccg ctgagcgctc
     1141 atgaccagct gaagcttgac aaggagaaac tccctgtcca tgtggttgtt gaccctattc
     1201 tcagtaaggt tttgcggcct catcagagag agggagtgaa attcctgtgg gagtgtgtca
     1261 ccagtcggcg catccctggc agccatggct gcatcatggc tgatgagatg ggcctaggaa
     1321 agacgctgca gtgcatcaca ttgatgtgga cacttttacg ccagagtcca gagtgcaagc
     1381 cagaaattga caaggcagtg gtggtgtcgc cttccagcct ggtgaagaac tggtacaatg
     1441 aggttgggaa atggctcgga gggaggatcc aacctctggc catcgatgga ggatctaagg
     1501 atgaaataga ccaaaagctg gaaggattca tgaaccagcg tggagccagg gtgtcttctc
     1561 ccatcctcat catttcctat gagaccttcc gccttcatgt tggagtcctc cagaaaggaa
     1621 gtgttggtct ggtcatatgt gacgagggac acaggctcaa gaactctgag aatcagactt
     1681 accaagccct ggacagcttg aacaccagcc ggcgggtgct catctccgga actcccatcc
     1741 agaatgatct gcttgagtat ttcagcttgg tacattttgt taattccggc atcctaggga
     1801 ctgcccatga attcaagaag cattttgaat tgccaatttt gaagggtcga gacgctgctg
     1861 ctagtgaggc agacaggcag ctaggagagg agcggctgcg ggagctcacc agcattgtga
     1921 atagatgcct gatacggagg acttctgata tcctttctaa atatctgcct gtgaagattg
     1981 agcaggtcgt ttgttgtagg ctgacacccc ttcagactga gttatacaag aggtttctga
     2041 gacaagccaa accggcagaa gaattgcttg agggcaagat gagtgtgtct tccctttctt
     2101 ccatcacctc gctaaagaag ctttgtaatc atccagctct aatctatgat aagtgtgtgg
     2161 aagaggagga tggctttgtg ggtgccttgg acctcttccc tcctggttac agctctaagg
     2221 ccctggagcc ccagctgtca ggtaagatgc tggtcctgga ttatattctg gcggtgaccc
     2281 gaagccgtag cagtgacaaa gtagtgctgg tgtcgaatta cacccagact ttggatctct
     2341 ttgagaagct gtgccgtgcc cgaaggtact tatacgtccg cctggatggc acgatgtcca
     2401 ttaagaagcg agccaaggtt gtagaacgct tcaatagtcc atcgagccct gactttgtct
     2461 tcatgctgag cagcaaagct gggggctgtg gcctcaatct cattggggct aaccggctgg
     2521 tcatgtttga ccctgactgg aacccagcca atgatgaaca agccatggcc cgggtctggc
     2581 gagatggtca aaagaagact tgctatatct accgcctgct gtctgcaggg accattgagg
     2641 agaagatctt ccagcgtcag agccacaaga aggcactgag cagctgtgtg gtggatgagg
     2701 agcaggatgt agagcgccac ttctctctgg gcgagttgaa ggagctgttt atcctggatg
     2761 aagctagcct cagtgacaca catgacaggt tgcactgccg acgttgtgtc aacagccgtc
     2821 agatccggcc accccctgat ggttctgact gcacttcaga cctggcaggg tggaaccact
     2881 gcactgataa gtgggggctc cgggatgagg tactccaggc tgcctgggat gctgcctcca
     2941 ctgccatcac cttcgtcttc caccagcgtt ctcatgagga gcagcggggc ctccgcggat
     3001 ccgactacaa agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg
     3061 atgacaagta gtgagggccc gatatctcga caatcaacct ctggattaca aaatttgtga
     3121 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     3181 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     3241 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     3301 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     3361 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     3421 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     3481 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     3541 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     3601 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     3661 cctttgggcc gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg gaaggagaca
     3721 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt
     3781 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     3841 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     3901 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     3961 agatctgcgc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     4021 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     4081 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4141 tctaaatcgg ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4201 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4261 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4321 actcaaccct atctcggtct attcttttga tttataaggg attttgggga tttcggccta
     4381 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     4441 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4501 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     4561 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     4621 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4681 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     4741 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     4801 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     4861 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     4921 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     4981 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     5041 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     5101 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     5161 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     5221 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     5281 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     5341 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     5401 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     5461 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     5521 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     5581 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     5641 ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     5701 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     5761 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     5821 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     5881 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     5941 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     6001 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     6061 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     6121 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     6181 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     6241 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     6301 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6361 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6421 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6481 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6541 tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt
     6601 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6661 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6721 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6781 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     6841 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6901 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6961 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     7021 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     7081 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     7141 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     7201 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     7261 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     7321 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     7381 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     7441 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     7501 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7561 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7621 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7681 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7741 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7801 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7861 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7921 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7981 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     8041 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     8101 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     8161 tggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     8221 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     8281 ggcttgaccg acaattgcat gaagaatctg cttagg
//
