LOCUS       V035782                 7839 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035782
VERSION     V035782
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"
     CDS             706..2565
                     /label="KLHL8(NM_020803)"
                     /note="KLHL8(NM_020803)"
                     /gene="KLHL8"
     CDS             2572..2595
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHHHH"
                     /label="8xHis"
     misc_feature    2615..3203
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3269..3317
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3519..3947
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3961..4290
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4357..5151
                     /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    5327..5448
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5497..5513)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5521..5537
                     /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(5545..5575)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5590..5611
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5899..6487)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6658..7518)
                     /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(7519..7623)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatggc ttcagattct
      721 atgagtagta aacaagctag gaatcacatt acaaagggga aaaggcaaca acagcaccag
      781 caaataaaga acagatcctc aattagtgat ggtgatggag aagattcctt tatttttgaa
      841 gcaaatgaag cttggaaaga ttttcatggt tctcttcttc gattttatga aaatggagaa
      901 ctctgtgatg tcacactcaa ggttggctca aagctaatct cttgtcacaa gctggtattg
      961 gcttgtgtta ttccctactt tagagccatg tttctttctg aaatggctga agccaagcaa
     1021 acgctgattg agattagaga ttttgatggt gatgcaatag aagacttggt aaagtttgtc
     1081 tattcttcac ggctcacttt gactgttgac aatgtccagc ctctcttata tgcagcctgt
     1141 attctgcagg ttgaactggt ggctagagct tgttgtgaat acatgaagtt acattttcat
     1201 ccctccaatt gcctggcagt aagagccttt gcagaaagtc acaatcgaat agacttaatg
     1261 gacatggcgg atcagtatgc ctgtgaccat tttactgaag tagtggagtg tgaagacttt
     1321 gtaagtgtat caccgcagca cctccataag cttttgtcct ccagtgatct aaatattgaa
     1381 aatgaaaagc aggtctataa tgctgccatc aagtggcttc ttgccaatcc tcagcatcat
     1441 tccaaatggt tggatgaaac acttgcacag gttcgcctgc cattgttgcc ggttgatttt
     1501 cttatgggtg ttgtggcaaa agaacagatt gtcaagcaaa atctaaaatg tagagattta
     1561 ctggatgaag caagaaatta ccaccttcac ttgagtagca gagcagtacc tgactttgaa
     1621 tactccattc ggactacccc aaggaagcat actgctggtg tgctgttttg tgtaggtggt
     1681 cgaggtggat ctggtgaccc ctttcgcagt attgaatgct attctatcaa caaaaacagt
     1741 tggttctttg gaccagaaat gaatagtcga aggcgacatg tgggtgtaat ctctgtggaa
     1801 ggtaaagtgt atgcagtagg tggacatgat ggaaatgaac atttagggag tatggagatg
     1861 tttgatcctc tcactaataa atggatgatg aaggcatcaa tgaacacaaa gaggcgagga
     1921 attgccttgg cttccttagg aggcccaatt tatgcaattg gagggttaga tgacaatact
     1981 tgcttcaatg atgtggagag atatgacata gaatctgatc agtggagtac agtggcacca
     2041 atgaatactc cccgtggagg agttggctct gttgctctag taaaccatgt ttatgcagta
     2101 ggtggcaatg atggaatggc ttctttatct agcgtggaga gatatgatcc acatctggat
     2161 aagtggatag aagttaaaga aatgggtcag cgaagagcag gcaatggagt tagcaagctt
     2221 catggttgct tatacgtagt tggtggtttt gatgataatt ctcctctgag ttcagttgag
     2281 cggtatgacc cccgaagcaa caagtgggat tatgtggcag cacttactac tcccagaggt
     2341 ggagtgggaa tcgcaacagt gatgggcaaa atctttgcag ttggtggtca taatggcaat
     2401 gcatacttaa atacagtaga agcgtttgat ccagtgctga ataggtggga gcttgttgga
     2461 tctgtgtctc actgcagagc tggagcagga gtagctgtgt gttcctgttt aactagccaa
     2521 attcgagatg taggtcatgg atccaataat gtggttgact gtatgctcga gcaccatcac
     2581 catcaccatc accattaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg
     2641 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     2701 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     2761 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     2821 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     2881 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     2941 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     3001 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     3061 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     3121 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     3181 ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag
     3241 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg
     3301 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     3361 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     3421 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     3481 agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     3541 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3601 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3661 agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3721 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3781 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3841 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc
     3901 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     3961 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4021 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     4081 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4141 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4201 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     4261 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     4321 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4381 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4441 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4501 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     4561 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4621 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4681 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4741 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4801 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4861 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     4921 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4981 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5041 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     5101 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5161 tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5221 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5281 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     5341 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5401 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     5461 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5521 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5581 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5641 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5701 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5761 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5821 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5881 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5941 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6001 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6061 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     6121 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6181 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6241 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6301 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     6361 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6421 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6481 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6541 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6601 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6661 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6721 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6781 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6841 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6901 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6961 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7021 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7081 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7141 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7201 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7261 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7321 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7381 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7441 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7501 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7561 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7621 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     7681 ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     7741 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     7801 caaggcttga ccgacaattg catgaagaat ctgcttagg
//
