LOCUS       V036630                 9948 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036630
VERSION     V036630
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..4628
                     /label="Rreb1(NM_001039188)"
                     /note="Rreb1(NM_001039188)"
                     /gene="Rreb1"
     CDS             4635..4700
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4724..5312
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5378..5426
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5628..6056
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6070..6399
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6466..7260
                     /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    7436..7557
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7606..7622)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7630..7646
                     /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(7654..7684)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7699..7720
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8008..8596)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8767..9627)
                     /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(9628..9732)
                     /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 ccaccatgac gtcgaattcg cccattggtt
      781 tagaaggctc agacctgtct tccatcaaca ccatgatgtc agcagtaatg agcgtagcga
      841 gtgtcacaga gaatggtggg agcccccagg gcatcaagtc ccccatgaaa cctccaggac
      901 caaatcggat tggcagaagg aaccaggaaa cgaaagagga gaagtcttcc tacaactgtc
      961 ccctatgtga aaagatttgc accacccagc accagctgac catgcacatc cgtcagcaca
     1021 acacagacac gggaggagct gaccacgcat gcagtatctg tgggaagtcg ctgagctcgg
     1081 ccagctccct ggatcgtcac atgctggtgc actctggcga gaggccttac aagtgtacag
     1141 tgtgtggcca gtctttcacc accaatggga acatgcacag acatatgaag attcatgaga
     1201 aggataccaa cagtactaca gctgcagccc ctccatcccc tctgaagcgc aggcggttgt
     1261 cctccaaaag gaagctgagt cacgatgccg agtcagaaga cccaggacca gctaaaaaga
     1321 tggtagaaga cgggcagtca ggcgatttgg acaagatgag cgatgaaatc tttcactgcc
     1381 cagtgtgttt caaggagttt gtttgcaagt atgaactgga gacccacatg gagacccact
     1441 cagataaccc actaagatgt gacatttgct gcgtcacctt ccgcacacat cgaggactgc
     1501 tgcgccacaa tgcacttgtc cacaagcagc ttcccagaga tgccatggga agacctttta
     1561 tccagaacaa cccttcgatt cctgctggct tccatgattt agggtttact gacttctcct
     1621 gtaggaagtt tcctcgaatc tctcaggcct ggtgtgagac aaacctacgg cggtgcatca
     1681 gcgagcagca ccggtttgtg tgtgacacct gcgacaaggc gttccccatg ctgtcgtcac
     1741 tcatcctgca caggcagagc cacatccctg ccgatcaggg acgggagaag ctccagacca
     1801 agaccctggc tgccgagtcc ttggaacaga aggccttcct ggccttgctg ggcctgcagc
     1861 acaccaaaga cgtcaagcct gccccggcgg aggagctcct gccggacgac aaccaagcaa
     1921 tacagctcca gacacttaag taccagctac ctcaggagcc tggctgcccc accgtgctga
     1981 gtgtgtctcc tcttgacgct gcttctttag gtggctctct gacagtcctc ccagctacca
     2041 aggagaatat gaagcatctg tccctgcagc ccttccagaa gggcttcatc atccagccag
     2101 acagtagcat tgtggttaag cctatttcag gagagtcggc cattgagctg gcagatatcc
     2161 aacagattct aaagatggca gcttccgctc ctccgcagat cagtcttccg ccactttcca
     2221 aggcccctgc caccccgctg caggcgattt tcaagcacat gcctcctttg aagccaaagc
     2281 ccttggtcac accccggaca gtggtagccg cctccacgcc cccacctctc atcaacgccc
     2341 agcaggcatc tcccggttgt atcagcccca gccttcctcc acagtccctg aagttcctca
     2401 aggggtcggt ggaggcagtg tccaatgttc atctgcttca gtccaagtct gggatccagc
     2461 caagcaccac cacacagctc ttcctgcagc aagctggagt ggagttgccg ggccagcccg
     2521 agatgaagac acagctggaa caagagagta tcattgaggc cctgctgccc ctcaacatgg
     2581 aggcgaagat caagcaggag ataacagaag gtgacctcaa ggccatcatg acaggcccca
     2641 gtggcaagaa gacccccgcc atgcgcaagg tgctctaccc ctgccgcttt tgcaaccagg
     2701 tgtttgcttt ctctggagtc ctgcgagccc acgttcgctc ccacctgggc atctcaccct
     2761 accaatgcaa catctgtgac tatattgctg cagacaaagc cgccctgatc cgtcacatcc
     2821 gcacacacag cggggaacgg ccttacatct gcaagatctg ccactaccca ttcacggtca
     2881 aagccaactg tgagcggcac ctgcgcaaga agcacctcaa ggccaccagg aaggacattg
     2941 agaagaacat agagtacgtg agcagcccca ccgcagagct ggtggacgcc ttctgcgccc
     3001 cagagaccgt gtgcagactg tgcggggagg atctgaagca ctaccgagct ctacgcatcc
     3061 acatgcgcac acactgcagc cggggcctgg gcggctgcca caaaggccgg aaacccttcg
     3121 agtgcaagga atgcaacgct cccttcgtgg ccaagcgcaa ctgcatccat cacatcctca
     3181 agcagcacct gcacgtgccc gagaaggaca tcgagagcta cgtgcttgcc accaacagtg
     3241 gcctcggccc cgcggacacg cccacggatg ccgcttccag aggagaagag ggcagctgtg
     3301 tcactttcgc ggagtgcaag cccctcgcca ctttcctgga gccccaaaat ggctttcttc
     3361 actcgagccc cacccagccc ctgccttccc acatctctgt caagctggag ccagccagca
     3421 gctttgccat ggacttcaac gaaccccttg acttttcgca gaaaggcctg gcactggtcc
     3481 aagtgaagca ggaaaatgtt tcctccttgc tgacgtcttc ctcctcctct gccctctatg
     3541 actgctccat ggagcccatt gacctgtcca tccccaagag cgtcaagaaa ggagacaagg
     3601 acacagttgt tcccagtgac gccaagaaac cggaaccgga agctgggcaa gccgagccgc
     3661 tctcaccccg cccaccaccc tgccctacct tgtcagtgac tgtggagccc aaggggagcc
     3721 tggaaacccc cacaggcacg gtggtggccg tcaccacagc tgccaagcta gagccccaca
     3781 ctcagcccct ccagggctcc gtgcagctgg ctgtccccat ctactccccc gcgctcgtca
     3841 gcaatactcc cctcttgggc aattctgctg ccctcttgaa caacccagcc ttgcttcggc
     3901 ccttacggcc aaagcccccc ctcctcttgc caaagccctc gatgacagag gagctgccgc
     3961 cactggcctc catcgcccag atcatttctt ccgtgtcctc ggcccctact ctgctgaaaa
     4021 cgaaggtagc tgaccctgga ccatcgatca ccagcagtaa cactgtggcc acagacagcc
     4081 caggaagctc catccccaaa gctgccgcca cccccactga caccacaagc tctaaagaat
     4141 ccagtgagcc accccctgca gccagcagcc ctgaggaagc cttgcctact gaacaagggc
     4201 cggctgccac ttcctcgtcg aggaagaggg gcaggaaaag gggactgaga aaccggcccc
     4261 tccccaacag cagtgctgtg gacctggact ccagtgggga gtttgctagc atcgagaaga
     4321 tgctggccac cacagatacc aacaagttca gtccctttct gcagactgca gaggatgaca
     4381 ctcaggaaga ggtggctgga gcccctgccg accagcatgg gcccgctgat gaggagcaag
     4441 gtagccccgc agaagacagg ctgctgagag caaagcggaa ctcctatgcc aactgcctgc
     4501 aaaagatcaa ctgtccccac tgtccccggg tcttcccttg ggccagctcc ctgcagcggc
     4561 acatgcttac acacactggt aagaaggccc tcacggctca ccaggcggtg agtcttgaaa
     4621 ggaaagaggg atccgactac aaagaccatg acggtgatta taaagatcat gacatcgact
     4681 acaaggatga cgatgacaag tagtgagggc ccgatatctc gacaatcaac ctctggatta
     4741 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     4801 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     4861 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     4921 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     4981 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     5041 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     5101 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     5161 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     5221 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     5281 gagtcggatc tccctttggg ccgcctcccc gcctggaaac gggggaggct aactgaaaca
     5341 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     5401 acgcacgggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     5461 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     5521 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     5581 ccctgccata gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag
     5641 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     5701 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     5761 tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca
     5821 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     5881 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     5941 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg
     6001 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     6061 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     6121 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     6181 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     6241 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     6301 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     6361 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     6421 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     6481 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     6541 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     6601 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     6661 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     6721 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     6781 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     6841 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     6901 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     6961 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     7021 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     7081 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     7141 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     7201 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     7261 gcgggactct ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     7321 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     7381 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     7441 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     7501 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     7561 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     7621 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     7681 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     7741 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     7801 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     7861 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     7921 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     7981 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     8041 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     8101 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     8161 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     8221 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     8281 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     8341 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     8401 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     8461 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     8521 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     8581 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     8641 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     8701 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     8761 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     8821 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     8881 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     8941 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     9001 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     9061 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     9121 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     9181 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     9241 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     9301 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     9361 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     9421 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     9481 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     9541 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     9601 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     9661 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     9721 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     9781 cccgatcccc tatggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag
     9841 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     9901 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
