LOCUS       V045707                 9309 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045707
VERSION     V045707
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..3989
                     /label="Krba1(NM_001347152)"
                     /note="Krba1(NM_001347152)"
                     /gene="Krba1"
     CDS             3996..4061
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4085..4673
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4739..4787
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4989..5417
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5431..5760
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5827..6621
                     /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    6797..6918
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6967..6983)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6991..7007
                     /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(7015..7045)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7060..7081
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7369..7957)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8128..8988)
                     /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(8989..9093)
                     /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 ccaccatggc gctccaggtg cccatcagct
      781 tcaaggactt ggctgtgcgg ttctctgaag aggaatggcg gctcctgcag gaggggcagc
      841 gggaattcta cagagacgtg atgcgggaaa actacgagac gttggtgtct gtggggacct
      901 ctgagctgct tcctctctct gctttcctgt cacctgcaga ggctggagga gccacatcag
      961 gagagagcca ccaggataag ggacaaaaac cccatttgga gcatagttcc cagggagagc
     1021 agcctcagca gagcctacac ctcaccgcat tagtgcagct ggtgaaggag attccagagt
     1081 tcttgtttgg agaagtgaag ggtactgagg actactccga gagtgggagc accagtctgg
     1141 atggggagca aacaagcccc gaggtagctg tggttgtgga agcttgccct ccccgaggcc
     1201 tgctcaattc tcttccggag agccctgcaa gccaccccag cctggccacc acacccacag
     1261 gcagctcaac ttccggtggc cctcctggag actgggcaca cggaagcccc ttacctgcta
     1321 taggaactga tgataagcca ttatctatag agaaggaagg tgtaggagcc tcgagagaga
     1381 cctccattca ttccactcaa agcctgggcc agagcaagag ttacctaaga caggagcgag
     1441 gcagcatggg aacaggaacc cttcctgaga acagcccatt gcaaggcctc atcaactgtt
     1501 tgaaggagat ccttgtgccc agaccccagc accgggggac agccccagac ttgccgcctt
     1561 ctctccctgg cttgagtgtg ttgaagcaga cgagagctga ggtagaggct gggagcctgc
     1621 cctgcccggt gaagacagag gcagcatccg gagattgtcc ccttcagggc ctgctgaact
     1681 gtctgaagga gatcccaaaa gctccagacc gacgtcccag cccctcagga gcatcagact
     1741 tgcagctgca ggaggaccca gggaaaagac attctggagg gatgagacac ctccagactc
     1801 ctccccaccc tagtcatgaa gctggcagta tgcttgccac ggtgaaggta gaagatggct
     1861 gggcccagag tcccccagtg ccagcatcct gccagcttag caggcaaggc tatagctcct
     1921 attccactgg agacaacaga gaggtccgtg tgccccgctg gggccccatg actctagcca
     1981 gcagggcctc aagctcaccc ctagaagctc tggaggcctg tctgaagggc atccctccag
     2041 gtgggtcatc acctcttcag tcactagcca tctcatggtc cagaagtcct cagctaggag
     2101 atgccgggtc tcagaggttt gagctacagc aacaaggatc tcacagtgaa gaagctacaa
     2161 gggagccact tctgcctctg agcttgcagg gctacatgag agaaggacct ggggttcaac
     2221 cctgtggctc ccagggtacc cctaccagct tctcctcagc cagcagcagt gatggggatc
     2281 tggatttcag gagccccagg agcagccagg ggcaacggct tgggaaaggc tatctaccag
     2341 gaaactctcc actccaaggc ctggagaact gcctgagaga gatccctatt cccaggccac
     2401 aggctgcctg gccatgctcc tcggctgtca acaggggatt gaagagaaca gagcccagga
     2461 actggactgg agacagagaa ggactgagag gtgaggcctc tgagccaccc cacctcagac
     2521 agcgtcctgg agaagtgccc agcaggagtc tgcatcaaga cagtccacag acctgtactt
     2581 ccacctgcca ccaagtgacc accaggccag gaacatggca atggccacaa gaggagacag
     2641 ccaccatgcc ctcccctctg caccgcctgg agaactctct gagggggatc ttgcctgtga
     2701 ggcccttgcg tttcacctgc gtgactggcc ctggccccag tcccagccct tgctccagct
     2761 ccagcttcag cagctctgat ggagaagacc taagaccaga gcctgcattt tggcagtcac
     2821 ccctccagca gaaagaccag cctccctcct gtaaggaccc tgttcgtctg tgccctgtct
     2881 ctggcgcatc tccaagagtc aacagcaata gctgttctgc ggaagaccgt gagagaacag
     2941 agcccaggga ctgcagcagc ctcagcgcag gaagagcaga agagaagccc cacccaccca
     3001 gaagagaaga tggtgcagag cgcacacgcc agcctgggcc tgtcaccaat gctgaaggaa
     3061 aaggagcagc agctggccac ccatcgcctg cccctcagct ggaggaaaag cctgagccca
     3121 agggtactga agactccagg gacctggagc ctggacatag accacccagt gccgcagcca
     3181 ggacccaagg gaagctgctc tctggagacc ctccggagtc acctagcaag tctccccttc
     3241 ccacaactgt cttgtcaaaa tggtcaccca cttctcttca gccaccatgc ccctgtggca
     3301 ggtccttgca gcaggagctg cataaccttg gtactgccct cacggacaag ctagaccggc
     3361 ttgcagcagc cttggcaggc ctgactcagg aagttgcaac catgaagacc caaatggatc
     3421 aactgcgaag gcacccacgg agccttggcc caaaaggtca gggttcctgg cagttggccc
     3481 tcccccagag acctcgctgg gtcaacagac tgggccacag acatctaccc tactggagac
     3541 agaagggccc caccaggccc agaccaaaga ttctgcggac ccaggcagaa ggctgcaaga
     3601 ctagtgaccg cccaggactc tctagaggga agggcagttt ggtgcctcag ctgcctccag
     3661 aggcttcctt ggtagaatct tccaggccca cctgtagttc atcccagcag atctcctcta
     3721 cgcctggagg ccacactgtg ctgactgcac accctcctct ggagcacact gcatgccacc
     3781 agaatcccct ctccccttca gtgcctactt cagtgcaggt cccccttgtg gcctcacctg
     3841 caaccagtgc agacacagaa cctccggctg ctagagtggc agccatcagc attccaaacc
     3901 agcccaagga acctgacagc ctgctagggg aagccctcag cagagacctc tggggaggtg
     3961 accaccggga cccaaggtgg ggggcccatg gatccgacta caaagaccat gacggtgatt
     4021 ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgatatct
     4081 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     4141 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     4201 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     4261 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     4321 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     4381 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     4441 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     4501 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     4561 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     4621 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa
     4681 cgggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     4741 gcaataaaaa gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt
     4801 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     4861 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     4921 agccaacgtc ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg
     4981 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     5041 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     5101 tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt
     5161 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     5221 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     5281 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     5341 tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca
     5401 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     5461 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     5521 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5581 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     5641 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     5701 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     5761 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     5821 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     5881 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     5941 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     6001 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     6061 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     6121 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     6181 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     6241 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     6301 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     6361 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     6421 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     6481 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     6541 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     6601 cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac
     6661 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     6721 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     6781 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     6841 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     6901 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     6961 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     7021 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     7081 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     7141 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     7201 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     7261 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     7321 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     7381 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     7441 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     7501 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     7561 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     7621 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     7681 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     7741 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     7801 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     7861 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     7921 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     7981 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     8041 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     8101 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     8161 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     8221 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     8281 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     8341 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     8401 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     8461 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     8521 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     8581 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     8641 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     8701 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     8761 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     8821 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     8881 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     8941 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     9001 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     9061 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     9121 cgtcgacgga tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct
     9181 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     9241 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     9301 ctgcttagg
//
