LOCUS       V029413                 8658 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029413
VERSION     V029413
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             927..3461
                     /label="PRMT9(NM_138364)"
                     /note="PRMT9(NM_138364)"
                     /gene="PRMT9"
     CDS             3501..4220
                     /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    4258..4482
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4528..4956
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4970..5299
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5366..6160
                     /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    6334..6455
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6504..6520)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6528..6544
                     /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(6552..6582)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6597..6618
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6906..7491)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7662..8522)
                     /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(8523..8627)
                     /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 gtttaaagcc actgggatcc gccaccatgt cgaactcgcg gcccaggtcc cgccgagacg
      961 ccgggggtgg cgctggggca gccggccggg acgagctggt gtcgcggtcc ttgcagagcg
     1021 cagagcactg tctgggcgtc caggacttcg gcactgccta tgcccactac ctcctcgtgc
     1081 tcagcctggc gccggagctg aaacacgacg tgaaggaaac ttttcagtac acacttttca
     1141 gatgggctga agagcttgat gctctcagtc ggatacaaga cttacttggt tgctatgagc
     1201 aggccttgga actgtttcct gatgatgaag tgatttgcaa tagtatgggg gagcatctct
     1261 tcagaatggg ctttagggat gaagcagctg ggtattttca taaagcagtg aagctaaacc
     1321 ctgatttcag tgatgcaaag gagaattttt atcgtgttgc aaactggttg gtggaacgct
     1381 ggcactttat catgcttaat gacaccaaga ggaatacaat ttataatgca gcaatccaaa
     1441 aggcagtttg tttggggtcc aaaagtgttt tggacattgg agcaggaact ggaatactaa
     1501 gcatgtttgc taaaaaagct ggagcacatt ccgtgtatgc ctgtgagtta tccaagacca
     1561 tgtatgaact tgcctgtgat gtcgtggcag caaacaagat ggaagcaggg atcaaactct
     1621 tacatacgaa gtcacttgac atagagattc caaaacatat tcccgaaaga gtgtccctag
     1681 ttgtaacaga aactgtcgat gcaggtttat ttggagaagg aattgtggag agtttgattc
     1741 atgcatggga gcatttactt ttacagccaa agaccaaagg tgaaagtgct aattgtgaaa
     1801 agtatgggaa agttatacca gcaagtgctg ttatatttgg gatggcagta gaatgtgcag
     1861 agataagaag acatcataga gtgggtatta aggacattgc tggtatccat ttgccaacaa
     1921 atgtgaaatt tcagagtccg gcttattctt ctgtagatac tgaagaaaca attgaacctt
     1981 atacaactga aaagatgagt cgagttcctg gaggatattt ggctttgaca gagtgctttg
     2041 aaattatgac agtagatttc aacaaccttc aggaattaaa aagtcttgca actaaaaagc
     2101 ctgataagat tggtattcct gttattaaag aaggcatact agatgctatt atggtttggt
     2161 ttgtgctcca gcttgatgat gaacatagtt tatccacaag tcctagtgag gaaacatgtt
     2221 gggaacaggc tgtctacccc gtacaggacc ttgcagacta ctggataaag cctggagacc
     2281 atgtgatgat ggaagtatct tgtcaagact gttacttaag aatccagagt attagtgtct
     2341 tgggtttgga atgtgaaatg gatgttgcaa aaagttttac ccagaataaa gacttgttat
     2401 cgttaggaaa tgaggctgaa ctttgtagtg ccctcgctaa ccttcagacc agtaaaccag
     2461 atgctgtaga gcagacatgt atattggaat ctacagaaat tgctttgctt aacaacatcc
     2521 catatcatga aggctttaaa atggcaatga gcaaagtttt gtcttcactg actccagaga
     2581 aactgtatca gaccatggat actcactgtc agaatgagat gagctctgga actggacaga
     2641 gtaatactgt acagaacatc cttgaacctt tctacgtgtt agatgtgtcc gaaggcttct
     2701 ctgttctgcc tgttattgct ggcacacttg ggcaggttaa accatacagt tctgtggaga
     2761 aagaccagca tcgtattgct ctggacctca tatctgaagc caatcacttt cctaaagaaa
     2821 cacttgagtt ttggctgaga catgtggagg atgaatctgc tatgttacaa aggccaaaat
     2881 cagacaagtt atggagcata attatattgg atgtcattga gccatctggg ctcattcagc
     2941 aggaaataat ggaaaaagct gcaatatcca ggtgtttact acaatctgga ggcaagatct
     3001 ttcctcagta tgtgctgatg tttgggttgc ttgtggaatc acagacactc ctagaggaga
     3061 atgctgttca aggaacagaa cgtactcttg gattaaatat agcacctttt attaaccagt
     3121 ttcaggtacc tatacgtgta tttttggacc tatcctcatt gccctgtata cctttaagca
     3181 agccagtgga actcttaaga ctagatttaa tgactccgta tttgaacacc tctaacagag
     3241 aagtaaaggt atacgtttgt aaatctggaa gactgactgc tattccattt tggtatcata
     3301 tgtaccttga tgaagagatt aggttggata cttcaagtga agcctcccac tggaaacaag
     3361 ctgcagttgt tttagataat cccatccagg ttgaaatggg agaggaactt gtactcagca
     3421 ttcagcatca caaaagcaat gtcagcatca cagtaaagca actctcgagt ggcggtggag
     3481 gaagcggcgg aggtggatct atggtgagca agggcgagga gctgttcacc ggggtggtgc
     3541 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     3601 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     3661 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     3721 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     3781 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     3841 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     3901 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     3961 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     4021 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     4081 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     4141 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     4201 tggacgagct gtacaagtag tctagagggc ccgtttaaac ccgctgatca gcctcgactg
     4261 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     4321 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     4381 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     4441 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     4501 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     4561 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     4621 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     4681 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     4741 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     4801 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     4861 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     4921 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     4981 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     5041 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     5101 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     5161 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     5221 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     5281 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     5341 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     5401 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     5461 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     5521 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     5581 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     5641 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     5701 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     5761 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     5821 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     5881 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     5941 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     6001 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     6061 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     6121 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     6181 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     6241 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     6301 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     6361 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     6421 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     6481 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     6541 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     6601 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     6661 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     6721 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     6781 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     6841 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     6901 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     6961 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     7021 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     7081 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     7141 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     7201 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     7261 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     7321 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     7381 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     7441 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     7501 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     7561 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     7621 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     7681 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     7741 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     7801 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     7861 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     7921 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     7981 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     8041 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     8101 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     8161 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     8221 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     8281 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     8341 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     8401 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     8461 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     8521 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     8581 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     8641 aaagtgccac ctgacgtc
//
