LOCUS       V046272                 7899 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046272
VERSION     V046272
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..2579
                     /label="Klhl10(NM_025727)"
                     /note="Klhl10(NM_025727)"
                     /gene="Klhl10"
     CDS             2586..2651
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2675..3263
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3329..3377
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3579..4007
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4021..4350
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4417..5211
                     /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    5387..5508
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5557..5573)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5581..5597
                     /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(5605..5635)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5650..5671
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5959..6547)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6718..7578)
                     /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(7579..7683)
                     /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 ccaccatgga gatggagagc accgctgcct
      781 ccacccggtt ccaccagcct cacatggaaa gaaagatgag tgcaatgacc tgtgagatct
      841 tcaacgagct ccggcttgag ggcaagctct gcgacgtggt catcaaagtc aatggcttcg
      901 agttcaatgc ccacaagaac atcctctgta gctgcagctc ctactttaga gctttgttta
      961 caagtggctg gaacaacact gagaagaagg tttacaacat tcctggcatt tcccccgaca
     1021 tgatgaagct aattattgag tatgcgtaca ctcggaccgt gcctatcacc cctgacaatg
     1081 tggagaagct gctggccgct gctgatcagt ttaacatcat gggcatcgtt aggggttgct
     1141 gtgagttcct caagtcggag ctgtgtctgg ataactgtat cggcatctgc aagttcacgg
     1201 actactacta ctgccccgag ctgaggcaga aggcctacat gttcatcctg cacaacttcg
     1261 aggagatggt gaaagtctcg gcagagtttc tggagctctc tgtcactgag ctcaaggaca
     1321 tcattgagaa agacgaactc aacgtgaagc aggaggacgc agtattcgag gccattttaa
     1381 agtggatttc tcacgacccc caaaatagga agcaacatat ttcagtttta cttcctaagg
     1441 ttcgcctggc cctaatgcat gctgagtact tcatgaataa tgttaagatg aatgactatg
     1501 tcaaagacag tgaggaatgt aagccagtca tcattaacgc cctaaaggct atgtatgacc
     1561 tgaacatgaa cggaccctcc aattctgact tcaccaaccc acttaccagg ccccgcctgc
     1621 cctatgccat cctattcgct attggtggct ggagtggcgg gagccccacc aatgccattg
     1681 aggcatacga tgctcgagca gacagatggg tgaatgtcac gtgtgaggaa gaaagtcctc
     1741 gagcctacca cggggcagcc tacttgaaag gctatgttta tatcatcggg gggtttgata
     1801 gcgtagacta cttcaatagt gttaagcgtt ttgaccctgt gaagaaaact tggcaccaag
     1861 tggcccctat gcactctaga cgttgctatg tcagcgtgac agtcctcagc aatttcatct
     1921 atgccatggg agggtttgat ggctacgtgc gtctcaacac tgctgaacgg tacgagccag
     1981 agaccaacca atggacactc attgccccca tgcatgagca gcggagcgat gcgagcgcca
     2041 caacactcta tgggaaggtc tacatatgtg gtgggtttaa cggaaacgaa tgcctgttta
     2101 cagcagaagt gtacaacact gagagcaatc aatggacagt cattgcaccc atgaggagca
     2161 ggaggagtgg aataggtgta attgcttacg gggaacatgt atacgcagtg ggtggctttg
     2221 atggagccaa tcgactgagg agtgcagaag cctacagccc agtggctaat acttggcgca
     2281 cgatccccac tatgtttaac cctcgtagca attttgggat cgaggtagtg gacgacctct
     2341 tgtttgtggt gggtggcttt aatggcttta ccaccacctt taacgttgag tgctatgatg
     2401 aaaagaccga tgagtggtat gacgctcatg acatgagcat ataccgcagt gctctgagct
     2461 gctgcgtggt accagggcta gccaatgttg gggagtatgc agctagacgg gacaacttca
     2521 cgggattagc actgcgagat gaagtaaagt attccgcttc gacaagtacc ctacctgtag
     2581 gatccgacta caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg
     2641 acgatgacaa gtagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg
     2701 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     2761 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     2821 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     2881 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     2941 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     3001 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     3061 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     3121 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     3181 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     3241 ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag
     3301 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg
     3361 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     3421 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     3481 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     3541 agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     3601 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3661 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3721 agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3781 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3841 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3901 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc
     3961 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     4021 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4081 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     4141 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4201 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4261 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     4321 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     4381 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4441 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4501 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4561 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     4621 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4681 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4741 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4801 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4861 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4921 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     4981 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5041 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5101 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     5161 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5221 tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5281 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5341 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     5401 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5461 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     5521 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5581 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5641 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5701 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5761 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5821 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5881 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5941 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6001 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6061 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6121 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     6181 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6241 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6301 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6361 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     6421 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6481 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6541 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6601 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6661 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6721 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6781 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6841 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6901 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6961 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7021 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7081 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7141 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7201 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7261 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7321 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7381 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7441 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7501 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7561 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7621 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7681 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     7741 ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     7801 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     7861 caaggcttga ccgacaattg catgaagaat ctgcttagg
//
