LOCUS       V022713                 6428 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022713
VERSION     V022713
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..1918
                     /label="Elavl1(NM_010485)"
                     /note="Elavl1(NM_010485)"
                     /gene="Elavl1"
     CDS             1925..1990
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2028..2252
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2298..2726
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2740..3069
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3136..3930
                     /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    4104..4225
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4274..4290)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4298..4314
                     /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(4322..4352)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4367..4388
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4676..5261)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5432..6292)
                     /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(6293..6397)
                     /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 atgtctaatg gttatgaaga
      961 ccacatggcg gaagactgca gggatgacat tgggagaacg aatttaattg tcaactacct
     1021 ccctcagaac atgacccaag aggaactacg aagtctgttc agcagcattg gcgaggttga
     1081 atctgcaaag cttattcggg ataaagtagc aggacacagc ttgggctacg gttttgtgaa
     1141 ctatgtgact gcaaaagatg cagagagagc aatcagcaca ctgaacggct tgagactcca
     1201 gtccaaaacc attaaggtgt catatgctcg cccaagctca gaggtcatca aagatgccaa
     1261 cttatacatc agtgggctcc caaggaccat gacacagaag gatgtggaag acatgttttc
     1321 tcggtttggg cgaatcatca actccagggt ccttgtggat cagaccacag gtttgtccag
     1381 aggggttgcc tttatccggt ttgacaaacg gtcagaagca gaagaggcaa ttaccagttt
     1441 caatggtcat aaacccccag gttcctccga gcccatcaca gtgaagtttg cagccaatcc
     1501 caaccagaac aaaaacatgg ctctcctctc gcagctgtac cactcgcctg ctaggcggtt
     1561 tggaggccct gtacaccacc aggcacagag attcaggttc tcccctatgg gtgtagatca
     1621 catgagtggg atttctggtg tcaatgtccc cggcaatgct tcctcgggct ggtgcatctt
     1681 catctacaac cttgggcaag acgccgatga ggggatcctc tggcagatgt ttggcccctt
     1741 tggtgcagtt accaatgtga aagtgattcg tgatttcaac accaacaagt gcaaagggtt
     1801 tggttttgtg accatgacaa actatgaaga agctgcaatg gccatagcaa gtctgaacgg
     1861 ctaccgcctg ggggacaaaa ttttacaggt ttccttcaaa accaacaagt cccacaaact
     1921 cgaggactac aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga
     1981 cgatgacaag tagtgagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
     2041 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     2101 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     2161 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     2221 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     2281 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     2341 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     2401 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     2461 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     2521 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     2581 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     2641 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     2701 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     2761 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     2821 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     2881 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     2941 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3001 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3061 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     3121 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3181 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3241 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3301 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3361 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     3421 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     3481 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     3541 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     3601 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     3661 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     3721 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     3781 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     3841 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     3901 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     3961 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     4021 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4081 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     4141 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     4201 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     4261 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     4321 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     4381 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     4441 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     4501 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     4561 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     4621 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     4681 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     4741 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4801 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4861 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4921 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4981 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5041 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5101 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5161 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg
     5221 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5281 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5341 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5401 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5461 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5521 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5581 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5641 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5701 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5761 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5821 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5881 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5941 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6001 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6061 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     6121 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6181 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6241 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6301 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6361 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6421 ctgacgtc
//
