LOCUS       V032195                 9999 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032195
VERSION     V032195
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"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             922..987
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             991..4812
                     /label="MEI1(NM_152513)"
                     /note="MEI1(NM_152513)"
                     /gene="MEI1"
     CDS             4849..5568
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK*"
                     /label="EGFP"
     polyA_signal    5599..5823
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5869..6297
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6311..6640
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6707..7501
                     /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    7675..7796
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7845..7861)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7869..7885
                     /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(7893..7923)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7938..7959
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8247..8832)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9003..9863)
                     /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(9864..9968)
                     /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 aagccaccat ggactacaaa gaccatgacg gtgattataa agatcatgac
      961 atcgactaca aggatgacga tgacaagctt atggctgtga ggcaggcggc gacggcgggc
     1021 actcccgggc ccaggagaga ggaagaggcg gcgcttctat tcgagagggc ccattaccgg
     1081 cacgacccgc gctggctgct gcccgtgacc ccccgcctgt gcctggcctg cgcgctggag
     1141 ctgctgccgg accccggcgt gtcgttagtg cgcaagaagc acatgttgtc ctgcttccaa
     1201 gatgcccttg tgaggcatac ctccctggtc acgcaactgg tgtctcagga tcagagagtc
     1261 tgcatccact tcataagtgt gctttttgga ctattatgca gcatggaaga tgggagtgtg
     1321 acagacctct gtattgaagt ccttattcag atcacaacgc agctgaagct ggagcagact
     1381 atccgttgcc tgctggatga gtgccacaaa gagctgtgta acatgccctc catgcgaggc
     1441 agcctggcca ccctgaccct tcttggcaag ttggtggatg ccatccctgc tctggcagac
     1501 gagcttgtaa tggagcatgg caacctgatg gagcatctgt tgagaggctt agtatacccc
     1561 agtgagggca tacaagcttc tgtctgttac ctttatggga agctatactc ctcaccagtg
     1621 gcagctgaga tgctttcagg acacttccgt gagaagcttt ttcccctctt cctttccatc
     1681 ctggatggtg cccagacaaa ggagctgcag attaactgct tgggtttgct gaggcagctg
     1741 ttgaagtatg atctctttgt gtccatgatc atgaaccagg atggactggg agaaagtgct
     1801 aagaatatcg aagggtcatc aggaaatacc tcactgcctt tggtgctcaa aaagcttctc
     1861 ctctctagag atgaaaccct gcaggtggcc agtgctcact gtataactgc ggtgcttgtc
     1921 cactccccag caaagcatgc gtcagccttc atccacgctg acatcccaga gttcctcttt
     1981 gagcatcttt cttcttccag tgaagtgctc gtctggtcca gctgtaactg cttgacactc
     2041 ctggtagaag agccactctt tttttccaag tgccacacgg tgtatgggat cgaggcagtg
     2101 gtgaggagcc tgcagggaag cctgaagatg aacaacatag agctgcacaa gcagggcctg
     2161 ctgcttttcg ctgaaatcct gacccggcag ccagaggaga tcaagctgtt cacaagctca
     2221 gccatgtgca gagatgctgg ccgtgccctc caagaagcag ttagcagccc tgtgctggag
     2281 gtggctgctg aggccttgaa ggccacttct gcttttctga ggaaggacca tcagagcact
     2341 ccacctgtgc agtatgggga actgcaggct ttgctagaag ccatgctaaa ccgatgtgcg
     2401 gagttttccc agaccttgct gagcaggagg cccctgggcc atgcctccag tagagattca
     2461 gagaaggcca ttcttcaaag gggaaagttc ctcctcagca ctctggaggg atttagaagt
     2521 gcctgcaggt tggctataga attccagagt gagccttcag cccaggagaa tccattcaca
     2581 gctcccagcg ccaagaagga agacaccttg gaggccttct cagaatttct tctcagtgcc
     2641 tgtgactcgc tgtgtatccc catggtgatg agacacttgg agcagaccac ccacccagct
     2701 ttgatggaag ttttcctctc aattctacat aacctctttg tcatcgttcc ccacatgaag
     2761 gagaagtttt ccaagaagct tgcttcctca tccttcatac gactgaccct ggagctgaag
     2821 gccaggtttt gcagtggtct gagtcactca gccctaaacc aggtgtgttc caatttcctc
     2881 tactatatgt gcctcaacct tctctcagct ccagagaaga caggaccacc ttccaaagaa
     2941 gaactctctg cagtgtctga gctcctgcag catgggctgc cccagataag cagcaggagc
     3001 cctgaaagcc ttgccttcct gtctgatcgc cagtacatgg agggagctgc tcgccagaga
     3061 cagtactgca tcctgctcct cttctacttg gcctacatcc atgaagacag gtttgtctca
     3121 gaggcagagt tatttgaggc tgtgcaaagc ttcctcctgt cgctgcagga ccagggcgag
     3181 cgccccccac tggtggtctt caaagcctcc atctatctgc ttgcaatctg ccaggacaaa
     3241 gacaatacac tacgtgagac tatggtcagt gcaatcagaa aattcctaga aggcatccca
     3301 gacctgcagc tagtctatac tcaccatccg ctcctgctca ggttctttct gttgtatcca
     3361 gagctcatga gtaggtatgg gcaccgtgtc ctggaacttt ggttcttctg ggaagagagc
     3421 agctatgagg aactggatga tgtcacctct gctgggcagc ccgcccttcc tgccagctta
     3481 gtagtcctgt tccagttgct cagaagcatc cccagcatcc tgctcatctt gctggacctc
     3541 atctattcca gcccagtgga cacagctcac aaggtactga ttagcctgag gaccttcctg
     3601 aggaggaatg aggatatcca agtgggcggt cttatccgag gccacttcct gctgatcctg
     3661 cagcgtctgc tagtggagca tggggcatcc ccatcaggag cctcggggaa cctaccattg
     3721 ctgctgagcc tcctctccct gatgcagctc aggaatgtgt cagagcaaga actggacagc
     3781 gtggccatga agctccttca ccaagtaagc aagctgtgtg ggaagtgcag ccccactgac
     3841 gtggacatcc tgcagccctc cttcaacttc ctgtattgga gccttcatca gaccacaccc
     3901 agcagtcaga aaagagctgc tgcagtgctc ctgagcagca caggcctgat ggagcttctg
     3961 gagaagatgc tggccctcac cttggcaaag gcagattctc ccaggactgc actcctctgc
     4021 tctgcctggc tgctcactgc ctccttctct gcccagcagc acaagggcag tttgcaggtt
     4081 caccagacac tctctgtgga aatggaccaa gtattgaagg ctctcagctt tccaaagaaa
     4141 aaggctgcac tactctcagc tgccatctta tgcttcctgc ggacagccct gcgacaaagc
     4201 ttttcctctg ccctggtagc cctggtgccc tcaggggccc agccactgcc agccaccaag
     4261 gacactgtcc tagctccact gcgaatgtcg caagtccggt ccctggtcat tgggctgcag
     4321 aacctcctgg tgcagaagga ccctctattg tcccaggcct gtgttggctg cctggaggcc
     4381 ttgcttgact acctggatgc ccggagccca gacattgctc tccacgtggc ctcccagcct
     4441 tggaatcggt ttttgctgtt taccctcttg gatgctggag agaattcctt cctcagacct
     4501 gagattttga ggctcatgac cctgtttatg cggtaccgga gtagcagtgt cctctctcat
     4561 gaagaggtgg gtgatgttct gcaaggtgtg gctttggctg acctgtctac cctctcgaac
     4621 accacactcc aggccctgca tggcttcttc cagcagctcc agagcatggg acacctggct
     4681 gaccacagca tggcccagac cctgcaggcc tccttggagg gccttccccc tagcacctcc
     4741 tcaggccagc cacccctgca ggacatgctc tgcctgggag gggtggctgt atccctgtcc
     4801 cacatcagaa actcgagtgg cggtggagga agcggcggag gtggatctat ggtgagcaag
     4861 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     4921 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     4981 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     5041 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     5101 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     5161 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     5221 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     5281 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     5341 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     5401 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     5461 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     5521 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaggg cccgtttaaa
     5581 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     5641 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     5701 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     5761 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     5821 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     5881 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     5941 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     6001 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     6061 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     6121 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     6181 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     6241 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     6301 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     6361 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     6421 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     6481 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     6541 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     6601 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     6661 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     6721 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     6781 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     6841 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     6901 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     6961 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     7021 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     7081 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     7141 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     7201 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     7261 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     7321 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     7381 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     7441 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     7501 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     7561 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     7621 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     7681 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     7741 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     7801 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     7861 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     7921 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     7981 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     8041 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     8101 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     8161 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     8221 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     8281 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     8341 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     8401 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     8461 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     8521 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     8581 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     8641 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     8701 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     8761 aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa
     8821 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     8881 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     8941 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     9001 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     9061 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     9121 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     9181 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     9241 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     9301 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     9361 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     9421 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     9481 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     9541 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     9601 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     9661 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     9721 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     9781 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     9841 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     9901 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     9961 gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
