LOCUS       V026036                 6911 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026036
VERSION     V026036
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      935..944
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             998..2293
                     /label="GFAP(NM_002055)"
                     /note="GFAP(NM_002055)"
                     /gene="GFAP"
     CDS             2408..2473
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2511..2735
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2781..3209
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3223..3552
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3619..4413
                     /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    4587..4708
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4757..4773)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4781..4797
                     /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(4805..4835)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4850..4871
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5159..5744)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5915..6775)
                     /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(6776..6880)
                     /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 atggagctgg gcctgagatg
      961 ggtgtttctg gtggccattc tggagggcgt gcagtgtatg gagaggagac gcatcacctc
     1021 cgctgctcgc cgctcctacg tctcctcagg ggagatgatg gtggggggcc tggctcctgg
     1081 ccgccgtctg ggtcctggca cccgcctctc cctggctcga atgccccctc cactcccgac
     1141 ccgggtggat ttctccctgg ctggggcact caatgctggc ttcaaggaga cccgggccag
     1201 tgagcgggca gagatgatgg agctcaatga ccgctttgcc agctacatcg agaaggttcg
     1261 cttcctggaa cagcaaaaca aggcgctggc tgctgagctg aaccagctgc gggccaagga
     1321 gcccaccaag ctggcagacg tctaccaggc tgagctgcga gagctgcggc tgcggctcga
     1381 tcaactcacc gccaacagcg cccggctgga ggttgagagg gacaatctgg cacaggacct
     1441 ggccactgtg aggcagaagc tccaggatga aaccaacctg aggctggaag ccgagaacaa
     1501 cctggctgcc tatagacagg aagcagatga agccaccctg gcccgtctgg atctggagag
     1561 gaagattgag tcgctggagg aggagatccg gttcttgagg aagatccacg aggaggaggt
     1621 tcgggaactc caggagcagc tggcccgaca gcaggtccat gtggagcttg acgtggccaa
     1681 gccagacctc accgcagccc tgaaagagat ccgcacgcag tatgaggcaa tggcgtccag
     1741 caacatgcat gaagccgaag agtggtaccg ctccaagttt gcagacctga cagacgctgc
     1801 tgcccgcaac gcggagctgc tccgccaggc caagcacgaa gccaacgact accggcgcca
     1861 gttgcagtcc ttgacctgcg acctggagtc tctgcgcggc acgaacgagt ccctggagag
     1921 gcagatgcgc gagcaggagg agcggcacgt gcgggaggcg gccagttatc aggaggcgct
     1981 ggcgcggctg gaggaagagg ggcagagcct caaggacgag atggcccgcc acttgcagga
     2041 gtaccaggac ctgctcaatg tcaagctggc cctggacatc gagatcgcca cctacaggaa
     2101 gctgctagag ggcgaggaga accggatcac cattcccgtg cagaccttct ccaacctgca
     2161 gattcgagaa accagcctgg acaccaagtc tgtgtcagaa ggccacctca agaggaacat
     2221 cgtggtgaag accgtggaga tgcgggatgg agaggtcatt aaggagtcca agcaggagca
     2281 caaggatgtg atgcccccag cagggactac cgatgctgcc caccctggta gaagcgtggt
     2341 tccagcactg ttacctctgc tggcaggtac tctgctgctg cttgaaactg ccacagcacc
     2401 cctcgaggac tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga
     2461 tgacgatgac aagtagtgag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     2521 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     2581 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     2641 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     2701 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     2761 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2821 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2881 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2941 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3001 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3061 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3121 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     3181 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     3241 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3301 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3361 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3421 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3481 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3541 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     3601 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     3661 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     3721 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     3781 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     3841 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     3901 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     3961 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4021 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     4081 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4141 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4201 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     4261 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4321 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     4381 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     4441 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     4501 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     4561 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     4621 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4681 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     4741 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     4801 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     4861 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4921 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     4981 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5041 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5101 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5161 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5221 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5281 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5341 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5401 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5461 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5521 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5581 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5641 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt
     5701 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     5761 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     5821 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     5881 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     5941 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6001 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6061 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6121 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6181 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     6241 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6301 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     6361 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     6421 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     6481 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     6541 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     6601 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     6661 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     6721 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     6781 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     6841 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     6901 cacctgacgt c
//
