LOCUS       V033568                 9039 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033568
VERSION     V033568
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..3719
                     /label="Nbr1(NM_008676)"
                     /note="Nbr1(NM_008676)"
                     /gene="Nbr1"
     CDS             3726..3791
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3815..4403
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4469..4517
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4719..5147
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5161..5490
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5557..6351
                     /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    6527..6648
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6697..6713)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6721..6737
                     /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(6745..6775)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6790..6811
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7099..7687)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7858..8718)
                     /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(8719..8823)
                     /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 accacaggtt actctaaatg
      781 tgacttttaa aaatgaaact caaagctttc tggtttcgga tccagaaaat acaacctggg
      841 ctgatgttga agctatggta aaagtttcat ttgatctgaa tactatccag ataaaatacc
      901 tggatgagga gaacgaggag atatccatca atagtcaagg tgaatatgaa gaagcactta
      961 agatggctaa cattaagcaa ggaaatcagc tacagatgca agtccacgaa gggtaccatg
     1021 ttgtagatga agcactcccc aaaaatgtgg tagaaaacca agcggctgcc aggacaggga
     1081 agaagccact tgcacattat tcttcactgg tgagagtctt gggatcagac atgaagacca
     1141 cagaggagcc tgcaccagag caatgttctt ctgctccttg tgacacagac cagcctcaag
     1201 acaagccccc agattggttt acaagctacc tggagatgtt ccgagaacaa gtggttaagg
     1261 aaacggtgga gaaacttgaa cagaggttgc aggagaagct tgtcctccag aagccactcc
     1321 tgagttcctc acccacagaa gtctcaatgc ctatttcaga ggaaaccctg tttttgccag
     1381 agaaccagtt cagctggcat attgcttgta gccactgcca gaagaggatc gttggagtgc
     1441 gctaccagtg tagcctatgc ccatcctaca acatttgtga agattgtgaa gctggaccat
     1501 acacccatga cactaaccat gttttgttaa agctgcggag acctgttgtg atatcctctg
     1561 agccattttt ctactccaag tattccgctc ctcgtctgcc tgctgctttg gaacaagtca
     1621 ggctccaaaa acaggtggac aagaactttg tgaaagcaga aaagcaaagg ttgcgggctg
     1681 agaagaaaca gcggaaggca gaggtcaaag agcttaaaaa gcagcttaaa ctccacagga
     1741 agattcattt atggaattca atccatggac ttcagagtcc caagtcccct ttgggccgac
     1801 ctgagagctt gctgcagtct aacaccctga tgcttccttt gcagccctgt gccccagtta
     1861 tgccaacact cagtgcagca tttgtggatg agaatttgcc tgatgggact catcttcagc
     1921 caggaaccaa gtttattaaa cactggagga tgaaaaacac ggggaatgtg aagtggaaca
     1981 cagacacaaa gctcaagttc atgtggggaa acttgactct ggcttctaca gagaagaaag
     2041 atgttttggt tccctgcttg aaggctggcc acgttggggt cgtgtctgtg gagttcatcg
     2101 ctccaacctt ggaggggaca tacacttcgc actggcgtct ctctcacaaa ggccagcagt
     2161 ttgggcctcg ggtttggtgc agtatcatag tggatccttt tccttcctct gagagccctg
     2221 ataatgttga aggggacagg atcagctcaa gcaaagctga cgatttctcc tgtgagcaag
     2281 aggaagcttt tcttctagct gaagaagaga ttccattggg tgaggtgaca aagcagacag
     2341 aagggacagg agccagtgcc tcacagaaga caagacgcgc tgccagtgaa agggagctgt
     2401 acattccatc tgtggacctc ctgactgccc aggacctgct gtcctttgag ctgctggata
     2461 taaacattgt ccaagaattg gagagagtgc cacacaacac ccctgtggat atgactccct
     2521 gcatgtctcc tctgccacat gacagtcctt taatagagaa gccaggcttg gggcagatac
     2581 aggaagagag tgaaggggcg ggatttaaag cacctcctga ttccactgta tcagcaaaga
     2641 ggaaagctga gacccctgct tcggtcgaag aaacagaaga agacctgagt gggacccaat
     2701 ttgtatgtga gactgtaatt cgatccctta ccttggatgc tgctccagac cacaaccctc
     2761 cttgcaggca gaggtcccca cagagggaat tacagctgta ttccaccgag ggacagcagc
     2821 cacttgtgct gcctggattc tgcagaaagg attcttcctt gaaatttgcc ttgcctgaag
     2881 aaggaccgcg tggagatgag agggaggaga ttgtccacat tgttgaggaa gaagtcgttg
     2941 aggaagagga ggaagtccaa gatgaagaag tccaaagtca gtcttctgct tcctctgaag
     3001 attatatcat catccttcct gagtgctttg ataccagccg ccccctgggg gactccatgt
     3061 acagctctgc gctctcccag ccaggcctgg agcgaggggc tgaaggcgaa cctgggatcg
     3121 agtctgggct ggaaccaact gaggctagag agagactccc agagagggag agccagccac
     3181 aggagcagag catcagcgac atcctcacaa cctcacagcc tctggacacg gtgcctctag
     3241 tccccgaggt ggcaggactt ccagcagcat tgtccaggag cgccccttgt ggacagtgtg
     3301 agtcatcagg agtggattca ccaggagtgg attcaccagc taccatgcac gaagttcccc
     3361 cagcccctga tgacatcaga ggagagccca gaggctcaac aggactcgca aacagcagac
     3421 agaggagctg tgaccactca aggcaccaca atgggagcag cattgctgga ggactggtga
     3481 agggggcttt gtctgttgct gcctctgcgt acaaggccct gttttctggg ccaccagtca
     3541 ctgcacagcc catcgtttct gaagatcaga ccacagccct gatggcccat ctctttgaaa
     3601 tgggattctg cgacaggcag ctgaacctaa ggctgctgag aaaacacaat tacaatatcc
     3661 tgcaggttgt gacagagctc cttcaagtga ataacaacga ctggtacagc caccgctatg
     3721 gatccgacta caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg
     3781 acgatgacaa gtagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg
     3841 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     3901 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     3961 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     4021 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     4081 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     4141 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     4201 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     4261 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     4321 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     4381 ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag
     4441 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg
     4501 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     4561 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     4621 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     4681 agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     4741 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     4801 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4861 agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4921 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4981 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     5041 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc
     5101 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     5161 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     5221 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     5281 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     5341 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     5401 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     5461 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     5521 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     5581 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     5641 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     5701 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     5761 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     5821 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     5881 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     5941 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     6001 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     6061 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     6121 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     6181 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     6241 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     6301 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     6361 tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     6421 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     6481 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     6541 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     6601 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     6661 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     6721 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     6781 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     6841 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     6901 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6961 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     7021 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     7081 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     7141 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     7201 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     7261 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     7321 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     7381 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     7441 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     7501 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     7561 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     7621 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     7681 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     7741 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     7801 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     7861 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7921 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7981 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     8041 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     8101 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     8161 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     8221 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     8281 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     8341 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     8401 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     8461 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     8521 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     8581 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     8641 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     8701 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     8761 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     8821 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     8881 ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     8941 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     9001 caaggcttga ccgacaattg catgaagaat ctgcttagg
//
