LOCUS       V045038                 7908 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045038
VERSION     V045038
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..2588
                     /label="Rhobtb3(NM_028493)"
                     /note="Rhobtb3(NM_028493)"
                     /gene="Rhobtb3"
     CDS             2595..2660
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2684..3272
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3338..3386
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3588..4016
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4030..4359
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4426..5220
                     /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    5396..5517
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5566..5582)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5590..5606
                     /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(5614..5644)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5659..5680
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5968..6556)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6727..7587)
                     /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(7588..7692)
                     /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 ccaccatgtc catccacatc gtggcgttgg
      781 ggaacgaggg cgacacgttc caccaggaca accggccgtc ggggctcatc cgcacatacc
      841 tgggaagaag cccgctggtg tcgggggacg agagtagctt gttgctgaac gcggccagca
      901 cggtcgcgcg cccggtgttc actgagtacc aggccagtgc gttcgggaac gtcaagctgg
      961 tggtgcacga ctgtcccgtc tgggacattt ttgacagtga ttggtacacc tcccgaaacc
     1021 taattggggg tgccgacatc attgtgatca aatacaatgt aaatgacaag ttttcattcc
     1081 atgaagtaaa ggataactat attccagtga ttaaaagagc atcaaactca gttccggtaa
     1141 tcattgcggc tgttggtacg agacaaaatg aagaattacc ttgtacatgc cccctgtgta
     1201 cttcagaccg agggagctgt gttaccacca cagaagggat ccaacttgcc aaagaactgg
     1261 gagctaccta tctggaactc cacagccttg atgatttcta cattggaaag tactttggag
     1321 gagtgctgga gtattttatg attcaagcct taaatcagaa gacaagtgaa aaaatgaaga
     1381 aaaggaaaat gaccagctcc ttccatggga ttcgacctcc tcagcttgaa cagccagaaa
     1441 agatgcctgt cttgaaagct gaggcatcac actatcactc cgacttgaat aacttgctgc
     1501 tctgctgcca gtgtgtggat gtggtttttt accacccaga agtgacgggc gtcgtggaag
     1561 cccacaagat cgttctctgc tctgtaagcc atgttttcat gctgcttttc aatgtgaaaa
     1621 gcccggccga cattcaggat tccagtatca tccgaaccac ccaggatctc tttgccataa
     1681 acagagatgc tgtgcttcca ggtgctagcc aggaagcccc cagcaaccca ccattgcctg
     1741 ttattgttaa ggacgccctc ttctgctcct gtttatcaga cattctgcgc ttcatttatt
     1801 caggtgcttt tcagtgggaa gagctggaag aagatgtcag gaggaagttg aaagattccg
     1861 gggatgtttc tgatataatc gagaaagtta aatgtatttt aaaaacacca ggaaagatta
     1921 attgcctgag gaactgcaag acctaccaag ccaggaagcc tctgtggttt tataatacgt
     1981 ctctcaagtt tttccttaat aaaccgatgc ttgccgatgt tgtctttgaa atacaaggag
     2041 caactgtgcc agcccacagg gccatcttgg tggcacgatg tgaagtgatg gctgccatgt
     2101 ttaatggtaa ttatatggaa gccaagagtg tactgattcc agtttatggc gtttccaaag
     2161 agacattctt gtcattttta gaatacttat acacagactc ttgttgccca gctggcattt
     2221 ttcaggccat gtgtctcctg atctgtgctg agatgtacca ggtctctaga ctgcagcaca
     2281 tctgtgagct gttcatcatc acacagctgc agagcatgcc cagcagggag ctggcctcca
     2341 tgaacctcga cattgtagac ctgctcaaaa aggccaagtt tcaccactca gactgccttt
     2401 ccacctggct gcttcatttc atcgccacta actacctcat attcagccaa aagcctgaat
     2461 ttcaagacct ttcagtggaa gaacgaagtt ttgttgaaaa gcacagatgg ccatcaaaca
     2521 tgtacttgaa gcagctggca gaatacagga agtatatcca ctcccggaaa tgtcgctgtt
     2581 tagtcatggg atccgactac aaagaccatg acggtgatta taaagatcat gacatcgact
     2641 acaaggatga cgatgacaag tagtgagggc ccgatatctc gacaatcaac ctctggatta
     2701 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     2761 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     2821 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     2881 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     2941 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     3001 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     3061 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     3121 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     3181 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     3241 gagtcggatc tccctttggg ccgcctcccc gcctggaaac gggggaggct aactgaaaca
     3301 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     3361 acgcacgggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     3421 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     3481 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     3541 ccctgccata gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag
     3601 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3661 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3721 tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca
     3781 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3841 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3901 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg
     3961 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     4021 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4081 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4141 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4201 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4261 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     4321 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     4381 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4441 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4501 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4561 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4621 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4681 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4741 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4801 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4861 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4921 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4981 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     5041 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     5101 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5161 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5221 gcgggactct ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     5281 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     5341 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     5401 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5461 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     5521 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     5581 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5641 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     5701 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     5761 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5821 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5881 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5941 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6001 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6061 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6121 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     6181 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6241 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6301 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6361 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     6421 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6481 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6541 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6601 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6661 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6721 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6781 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6841 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6901 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6961 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     7021 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     7081 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     7141 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7201 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7261 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7321 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7381 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7441 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7501 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7561 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7621 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7681 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     7741 cccgatcccc tatggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag
     7801 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     7861 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
