LOCUS       V045663                 8262 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045663
VERSION     V045663
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..2942
                     /label="Arhgap28(NM_172964)"
                     /note="Arhgap28(NM_172964)"
                     /gene="Arhgap28"
     CDS             2949..3014
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3038..3626
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3692..3740
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3942..4370
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4384..4713
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4780..5574
                     /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    5750..5871
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5920..5936)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5944..5960
                     /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(5968..5998)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6013..6034
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6322..6910)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7081..7941)
                     /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(7942..8046)
                     /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 ggtggaggac tcgggcggcg
      781 tggtgctgac cgcctaccac tcgcacgcgc gctcgcaacc gcagggcgct gagccacgct
      841 gcgcgtcccg ggccagccat ccgctcagca gaaaatccat tcctcgctgc cgacgaatca
      901 acaggatgct ctccaatgaa tccctgcatc ctccctcctt tagtcgctcc aactctcaag
      961 cctcagtaga cagcagcgca tccatggagg agttcctgcg ggaaattgag agtatcaaag
     1021 agagcagcgt gggggcgtcg caggagcagc cacccactgc tgctgctgct gctgctgaag
     1081 tcaagccagt ggatgaagga gagcttgaag cggagtggct acaagatgtg ggattgtcaa
     1141 ctctgatctc tggcaacgaa gaggaagacg gtaaagcgct cctgtctaca ttgactcgga
     1201 cccaagctgc tgcagtgaaa aagagataca acacatatac tcagactctg aggaaaaaga
     1261 ataaacaacc cgtcagggat gtcagagaca tcttcggagt cagcgaatct cctcctagtg
     1321 attcttgtga gcatgctact cagttggacg gtaccaagga agaaaaagat ctgccaggag
     1381 ttaccaagac aagcagacct ctgccagacg atgcttctct cagtagtacc accctatcca
     1441 atggcgccca ggatgaagaa ggtggttttg tggccctaca gagtggttct gtgtcaatac
     1501 ttgaggctat cccggatatt gctgttcaca ccaatggatc tgcagatgct gagcagtcgg
     1561 ttcagagcac attgagtgat gatgattatc acgggaagaa cgttccagca gaggctgaag
     1621 agctgtcctt tgaggtctct tactcagaaa tggtgacaga aatgcctgat agaaataaat
     1681 ggaagaagtc ggacattaag aaagaagact atgtcttaac taaatttatc attcagaaaa
     1741 caagatttgg gttgaccgag acgggggacc tgtctgtgga agacatgaag aaaatccgcc
     1801 acctctctct gattgagctg acagccttct tcgatgcttt tgggatacaa ctgaagagaa
     1861 acaaaaccga gcgagtgaga ggccgagaca acgggatttt cggagtgcca cttacagtcc
     1921 tcctggacaa tgaccggaag aaggaccctg cagtgaaagt tcccctggtg ttacaaaaat
     1981 tttttcagaa agtcgaggaa tcaggtctgg aatcggaagg aatttttcga ctctcgggat
     2041 gtactgccaa agtcaagcag taccgcgaag aactggatgc caggttcaat gctgacaagt
     2101 tcaagtggga caagatgtgt cacagagaag ctgcggtcat gttgaaagca ttcttcagag
     2161 agctgcccac ctctctcttc cctgttgaat atatacccgc cttcatctct ctgatggaaa
     2221 gaggacctga catcaaagtg cagtttcaag ccttgcacct catggtcatg gccctgcctg
     2281 atgccaacag ggacacagcg caggctctga tggcattctt caataaagtg atcgccaacg
     2341 aatccaagaa ccgcatgaac ctgtggaaca tttccacagt gatggcgccc aacttgttct
     2401 tcagcagaag caagcactct gactgcgagg aactgctgct ggccaacacg gcgacccaca
     2461 tcatccgcct gatgctgaag taccagaaga ttctgtggaa ggttccatct ttcttgatca
     2521 cccaggtcag aaggatgaat gaggccacca tgctgttgaa gaagcagctc ccgagtatga
     2581 agaagctgct caggaggaag accctggatc gagaggtttc aatcctcaag acctcaaagg
     2641 taccacaaaa atcaccttcc tcaagaagaa tgtctgatgt gccagaaggt gtcataaggg
     2701 tccacgctcc acttctgtcc aaagtgtcca tggccattca gctcaacagt cagaccaaag
     2761 ccaaagatat cctggcgaaa tttcagtatg agaacagtca tggttcatct gaacatatta
     2821 agatgcagaa ccaaaggtta tatgaagttg gaggaaatat aggacaacac tgtttagatc
     2881 ctgatgcata tatattggat gtgtatcata taaatcctca tgcagaatgg gtcatcaagc
     2941 ccggatccga ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg
     3001 atgacgatga caagtagtga gggcccgata tctcgacaat caacctctgg attacaaaat
     3061 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     3121 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     3181 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     3241 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     3301 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     3361 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     3421 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
     3481 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     3541 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     3601 gatctccctt tgggccgcct ccccgcctgg aaacggggga ggctaactga aacacggaag
     3661 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     3721 gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     3781 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     3841 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     3901 catagcagat ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     3961 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4021 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4081 tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga
     4141 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4201 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4261 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tggggatttc
     4321 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     4381 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4441 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4501 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4561 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4621 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     4681 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     4741 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     4801 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     4861 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     4921 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     4981 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     5041 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     5101 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     5161 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     5221 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     5281 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     5341 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     5401 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     5461 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     5521 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     5581 ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     5641 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     5701 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     5761 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5821 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     5881 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     5941 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     6001 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     6061 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     6121 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     6181 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6241 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6301 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6361 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6421 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6481 cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag
     6541 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6601 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6661 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6721 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     6781 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6841 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6901 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6961 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     7021 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     7081 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     7141 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7201 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     7261 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7321 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7381 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7441 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7501 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7561 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7621 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7681 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7741 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7801 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7861 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7921 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7981 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     8041 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     8101 cccctatggt cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     8161 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     8221 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gg
//
