LOCUS       V047204                 7431 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047204
VERSION     V047204
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..2111
                     /label="Glra2(NM_183427)"
                     /note="Glra2(NM_183427)"
                     /gene="Glra2"
     CDS             2118..2183
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2207..2795
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2861..2909
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3111..3539
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3553..3882
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3949..4743
                     /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    4919..5040
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5089..5105)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5113..5129
                     /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(5137..5167)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5182..5203
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5491..6079)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6250..7110)
                     /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(7111..7215)
                     /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 ccaccatgta ccggcagcta gtgaacattt
      781 tgacagcctt gtttgcattt ttcttaggga caaaccactt cagggaagct ttctgcaaag
      841 accatgactc caggtctgga aaacatccct cgcagaccct atctccttca gatttcttgg
      901 ataaattaat gggaagaaca tcaggatatg atgcaagaat caggccaaat tttaaaggtc
      961 ctccagtaaa cgttacttgc aatattttta tcaacagttt tggatcagtc acagaaacca
     1021 ccatggacta ccgagtgaac atttttctga gacagcagtg gaatgattca cggctggcat
     1081 acagtgagta cccagatgat tccctggatt tggatccctc aatgttggat tcgatttgga
     1141 aaccggattt gttctttgcc aatgaaaagg gtgccaattt ccatgatgtc accactgaca
     1201 acaagttgtt gcggatttcc aaaaatggca aagtgctcta cagtattaga ctcaccttga
     1261 ctttatcttg tccaatggac ctgaagaact ttccaatgga tgtccagacc tgtacaatgc
     1321 agctggagag ttttgggtac accatgaatg acctgatatt tgagtggtta agtgatggtc
     1381 cagtacaagt tgctgaagga ctcaccttgc cccagtttat tttgaaagaa gagaaggagc
     1441 ttggttattg cacaaagcat tacaacactg gcaagtttac ctgcattgag gtcaagtttc
     1501 acctggagcg ccagatgggc tactatttga tccagatgta catccccagc ctgttgatag
     1561 tcattttgtc ctgggtctcc ttttggataa acatggatgc agcccctgcc agggttgccc
     1621 tgggcatcac aacagtcctg acaatgacta cacagagttc aggttccagg gcatctctgc
     1681 caaaggtctc ctatgtgaaa gcaattgaca tctggatggc ggtatgcctt ctctttgtgt
     1741 ttgctgcctt actggaatat gcagcagtga actttgtctc caggcaacat aaggagttcc
     1801 ttcgtctccg gagacgacag aagaggcaga ataaggaaga agatgttact cgtgaaagtc
     1861 gttttaactt cagtggctat gggatgggtc actgcctcca aatgaaagat ggcacagctg
     1921 tcaaggctac acctgccaac ccacttccac aaccccccaa agatgcagat gctatcaaga
     1981 agaagtttgt ggatcgggca aaaagaattg acaccatatc tcgagctgcc ttcccactgg
     2041 ccttcctcat tttcaacatc ttttactgga tcacatacaa gatcattcgg catgaagatg
     2101 tccacaagaa aggatccgac tacaaagacc atgacggtga ttataaagat catgacatcg
     2161 actacaagga tgacgatgac aagtagtgag ggcccgatat ctcgacaatc aacctctgga
     2221 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     2281 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     2341 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     2401 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     2461 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     2521 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     2581 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     2641 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     2701 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     2761 gacgagtcgg atctcccttt gggccgcctc cccgcctgga aacgggggag gctaactgaa
     2821 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     2881 aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     2941 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     3001 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     3061 aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc acgcgccctg
     3121 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3181 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3241 ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg
     3301 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3361 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3421 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3481 ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     3541 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3601 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3661 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3721 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3781 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     3841 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     3901 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     3961 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4021 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4081 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     4141 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4201 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4261 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4321 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4381 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4441 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     4501 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4561 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4621 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4681 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4741 tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4801 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4861 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     4921 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4981 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5041 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     5101 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     5161 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     5221 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5281 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5341 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5401 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5461 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5521 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5581 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5641 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     5701 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5761 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5821 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5881 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     5941 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6001 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6061 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6121 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6181 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6241 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6301 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6361 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6421 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6481 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6541 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6601 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6661 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6721 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6781 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6841 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6901 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6961 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7021 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7081 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7141 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7201 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     7261 tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc atagttaagc
     7321 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     7381 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag g
//
