LOCUS       V023780                 7892 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023780
VERSION     V023780
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             941..3382
                     /label="ARHGAP26(NM_015071)"
                     /note="ARHGAP26(NM_015071)"
                     /gene="ARHGAP26"
     CDS             3389..3454
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3492..3716
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3762..4190
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4204..4533
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4600..5394
                     /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    5568..5689
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5738..5754)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5762..5778
                     /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(5786..5816)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5831..5852
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6140..6725)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6896..7756)
                     /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(7757..7861)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atggggctcc cagcgctcga
      961 gttcagcgac tgctgcctcg atagtccgca cttccgagag acgctcaagt cgcacgaagc
     1021 agagctggac aagaccaaca aattcatcaa ggagctcatc aaggacggga agtcactcat
     1081 aagcgcgctc aagaatttgt cttcagcgaa gcggaagttt gcagattcct taaatgaatt
     1141 taaatttcag tgcataggag atgcagaaac agatgatgag atgtgtatag caagatcttt
     1201 gcaggagttt gccactgtcc tcaggaatct tgaagatgaa cggatacgga tgattgagaa
     1261 tgccagcgag gtgctcatca ctcccttgga gaagtttcga aaggaacaga tcggggctgc
     1321 caaggaagcc aaaaagaagt atgacaaaga gacagaaaag tattgtggca tcttagaaaa
     1381 acacttgaat ttgtcttcca aaaagaaaga atctcagctt caggaggcag acagccaagt
     1441 ggacctggtc cggcagcatt tctatgaagt atccctggaa tatgtcttca aggtgcagga
     1501 agtccaagag agaaagatgt ttgagtttgt ggagcctctg ctggccttcc tgcaaggact
     1561 cttcactttc tatcaccatg gttacgaact ggccaaggat ttcggggact tcaagacaca
     1621 gttaaccatt agcatacaga acacaagaaa tcgctttgaa ggcactagat cagaagtgga
     1681 atcactgatg aaaaagatga aggagaatcc ccttgagcac aagaccatca gtccctacac
     1741 catggaggga tacctctacg tgcaggagaa acgtcacttt ggaacttctt gggtgaagca
     1801 ctactgtaca tatcaacggg attccaaaca aatcaccatg gtaccatttg accaaaagtc
     1861 aggaggaaaa gggggagaag atgaatcagt tatcctcaaa tcctgcacac ggcggaaaac
     1921 agactccatt gagaagaggt tttgctttga tgtggaagca gtagacaggc caggggttat
     1981 caccatgcaa gctttgtcgg aagaggaccg gaggctctgg atggaagcca tggatggccg
     2041 ggaacctgtc tacaactcga acaaagacag ccagagtgaa gggactgcgc agttggacag
     2101 cattggcttc agcataatca ggaaatgcat ccatgctgtg gaaaccagag ggatcaacga
     2161 gcaagggctg tatcgaattg tgggtgtcaa ctccagagtg cagaagttgc tgagtgtcct
     2221 gatggacccc aagactgctt ctgagacaga aacagatatc tgtgctgaat gggagataaa
     2281 gaccatcact agtgctctga agacctacct aagaatgctt ccaggaccac tcatgatgta
     2341 ccagtttcaa agaagtttca tcaaagcagc aaaactggag aaccaggagt ctcgggtctc
     2401 tgaaatccac agccttgttc atcggctccc agagaaaaat cggcagatgt tacagctgct
     2461 catgaaccac ttggcaaatg ttgctaacaa ccacaagcag aatttgatga cggtggcaaa
     2521 ccttggtgtg gtgtttggac ccactctgct gaggcctcag gaagaaacag tagcagccat
     2581 catggacatc aaatttcaga acattgtcat tgagatccta atagaaaacc acgaaaagat
     2641 atttaacacc gtgcccgata tgcctctcac caatgcccag ctgcacctgt ctcggaagaa
     2701 gagcagtgac tccaagcccc cgtcctgcag cgagaggccc ctgacgctct tccacaccgt
     2761 tcagtcaaca gagaaacagg aacaaaggaa cagcatcatc aactccagtt tggaatctgt
     2821 ctcatcaaat ccaaacagca tccttaattc cagcagcagc ttacagccca acatgaactc
     2881 cagtgaccca gacctggctg tggtcaaacc cacccggccc aactcactcc ccccgaatcc
     2941 aagcccaact tcacccctct cgccatcttg gcccatgttc tcggcgccat ccagccctat
     3001 gcccacctca tccacgtcca gcgactcatc ccccgtcagg tctgttgcag ggtttgtttg
     3061 gttttctgtt gctgccgttg ttctctcatt ggctcggtcc tctcttcatg cagtgttcag
     3121 cctcctcgtc aactttgttc cctgccatcc aaacctgcac ttgctttttg acaggccaga
     3181 agaagcggta catgaagact ccagcacacc gttccggaag gcaaaagcct tgtatgcctg
     3241 caaagctgaa catgactcag aactttcgtt cacagcaggc acggtcttcg ataacgttca
     3301 cccatctcag gagcctggct ggttggaggg gactctgaac ggaaagactg gcctcatccc
     3361 tgagaattac gtggagttcc tcctcgagga ctacaaagac catgacggtg attataaaga
     3421 tcatgacatc gactacaagg atgacgatga caagtagtga gggcccgttt aaacccgctg
     3481 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     3541 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     3601 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     3661 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     3721 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     3781 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3841 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3901 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3961 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4021 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4081 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     4141 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     4201 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4261 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4321 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4381 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4441 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     4501 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     4561 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     4621 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     4681 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     4741 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     4801 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     4861 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     4921 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     4981 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     5041 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     5101 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     5161 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     5221 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     5281 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     5341 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     5401 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     5461 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     5521 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     5581 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     5641 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     5701 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     5761 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     5821 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     5881 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     5941 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     6001 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     6061 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6121 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6181 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6241 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6301 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6361 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6421 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6481 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6541 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     6601 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6661 ccaccgctgg tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6721 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6781 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     6841 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     6901 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6961 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     7021 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     7081 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     7141 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     7201 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     7261 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     7321 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     7381 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     7441 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7501 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     7561 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7621 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7681 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7741 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     7801 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     7861 gcacatttcc ccgaaaagtg ccacctgacg tc
//
