LOCUS       V027674                 9006 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027674
VERSION     V027674
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..3686
                     /label="ERN1(NM_001433)"
                     /note="ERN1(NM_001433)"
                     /gene="ERN1"
     CDS             3693..3758
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3782..4370
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4436..4484
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4686..5114
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5128..5457
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5524..6318
                     /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    6494..6615
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6664..6680)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6688..6704
                     /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(6712..6742)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6757..6778
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7066..7654)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7825..8685)
                     /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(8686..8790)
                     /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 ccaccatgcc ggcccggcgg ctgctgctgc
      781 tgctgacgct gctgctgccc ggcctcggga tttttggaag taccagcaca gtgacgcttc
      841 ctgaaacctt gttgtttgtg tcaacgctgg atggaagttt gcatgctgtc agcaagagga
      901 caggctcaat caaatggact ttaaaagaag atccagtcct gcaggtccca acacatgtgg
      961 aagagcctgc ctttctccca gatcctaatg atggcagcct gtatacgctt ggaagcaaga
     1021 ataatgaagg cctgacgaaa cttcctttta ccatcccaga attggtgcag gcatccccat
     1081 gccgaagttc agatggaatc ctctacatgg gtaaaaagca ggacatctgg tatgttattg
     1141 acctcctgac cggagagaag cagcagactt tgtcatcggc ctttgcagat agtctctgcc
     1201 catcaacctc tcttctgtat cttgggcgaa cagaatacac catcaccatg tacgacacca
     1261 aaacccgaga gctccggtgg aatgccacct actttgacta tgcggcctca ctgcctgagg
     1321 acgacgtgga ctacaagatg tcccactttg tgtccaatgg tgatgggctg gtggtgactg
     1381 tggacagtga atctggggac gtcctgtgga tccaaaacta cgcctcccct gtggtggcct
     1441 tttatgtctg gcagcgggag ggtctgagga aggtgatgca catcaatgtc gctgtggaga
     1501 ccctgcgcta tctgaccttc atgtctgggg aggtggggcg catcacaaag tggaagtacc
     1561 cgttccccaa ggagacagag gccaagagca agctgacgcc cactctgtat gttgggaaat
     1621 actctaccag cctctatgcc tctccctcaa tggtacacga gggggttgct gtcgtgcccc
     1681 gcggcagcac acttcctttg ctggaagggc cccagactga tggcgtcacc attggggaca
     1741 agggggagtg tgtgatcacg cccagcacgg acgtcaagtt tgatcccgga ctcaaaagca
     1801 agaacaagct caactacttg aggaattact ggcttctgat aggacaccat gaaaccccac
     1861 tgtctgcgtc taccaagatg ctggagagat ttcccaacaa tctacccaaa catcgggaaa
     1921 atgtgattcc tgctgattca gagaaaaaga gctttgagga agttatcaac ctggttgacc
     1981 agacttcaga aaacgcacct accaccgtgt ctcgggatgt ggaggagaag cccgcccatg
     2041 cccctgcccg gcccgaggcc cccgtggact ccatgcttaa ggacatggct accatcatcc
     2101 tgagcacctt cctgctgatt ggctgggtgg ccttcatcat cacctatccc ctgagcatgc
     2161 atcagcagca gcagctccag caccagcagt tccagaagga actggagaag atccagctcc
     2221 tgcagcagca gcagcagcag ctgcccttcc acccacctgg agacacggct caggacggcg
     2281 agctcctgga cacgtctggc ccgtactcag agagctcggg caccagcagc cccagcacgt
     2341 cccccagggc ctccaaccac tcgctctgct ccggcagctc tgcctccaag gctggcagca
     2401 gcccctccct ggaacaagac gatggagatg aggaaaccag cgtggtgata gttgggaaaa
     2461 tttccttctg tcccaaggat gtcctgggcc atggagctga gggcacaatt gtgtaccggg
     2521 gcatgtttga caaccgcgac gtggccgtga agaggatcct ccccgagtgt tttagcttcg
     2581 cagaccgtga ggtccagctg ttgcgagaat cggatgagca cccgaacgtg atccgctact
     2641 tctgcacgga gaaggaccgg caattccagt acattgccat cgagctgtgt gcagccaccc
     2701 tgcaagagta tgtggagcag aaggactttg cgcatctcgg cctggagccc atcaccttgc
     2761 tgcagcagac cacctcgggc ctggcccacc tccactccct caacatcgtt cacagagacc
     2821 taaagccaca caacatcctc atatccatgc ccaatgcaca cggcaagatc aaggccatga
     2881 tctccgactt tggcctctgc aagaagctgg cagtgggcag acacagtttc agccgccgat
     2941 ctggggtgcc tggcacagaa ggctggatcg ctccagagat gctgagcgaa gactgtaagg
     3001 agaaccctac ctacacggtg gacatctttt ctgcaggctg cgtcttttac tacgtaatct
     3061 ctgagggcag ccaccctttt ggcaagtccc tgcagcggca ggccaacatc ctcctgggtg
     3121 cctgcagcct tgactgcttg cacccagaga agcacgaaga cgtcattgca cgtgaattga
     3181 tagagaagat gattgcgatg gatcctcaga aacgcccctc agcgaagcat gtgctcaaac
     3241 acccgttctt ctggagccta gagaagcagc tccagttctt ccaggacgtg agcgacagaa
     3301 tagaaaagga atccctggat ggcccgatcg tgaagcagtt agagagaggc gggagagccg
     3361 tggtgaagat ggactggcgg gagaacatca ctgtccccct ccagacagac ctgcgtaaat
     3421 tcaggaccta taaaggtggt tctgtcagag atctcctccg agccatgaga aataagaagc
     3481 accactaccg ggagctgcct gcagaggtgc gggagacgct ggggtccctc cccgacgact
     3541 tcgtgtgcta cttcacatct cgcttccccc acctcctcgc acacacctac cgggccatgg
     3601 agctgtgcag ccacgagaga ctcttccagc cctactactt ccacgagccc ccagagcccc
     3661 agcccccagt gactccagac gccctcggat ccgactacaa agaccatgac ggtgattata
     3721 aagatcatga catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga
     3781 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     3841 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     3901 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     3961 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     4021 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     4081 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     4141 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     4201 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     4261 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     4321 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg
     4381 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     4441 ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg
     4501 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     4561 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     4621 caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta
     4681 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     4741 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     4801 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg
     4861 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     4921 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4981 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     5041 tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     5101 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     5161 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     5221 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     5281 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     5341 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     5401 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     5461 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     5521 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     5581 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     5641 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     5701 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     5761 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     5821 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     5881 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     5941 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     6001 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     6061 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     6121 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     6181 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     6241 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     6301 cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc
     6361 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     6421 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     6481 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     6541 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     6601 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     6661 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     6721 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     6781 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     6841 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     6901 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     6961 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     7021 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     7081 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     7141 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     7201 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     7261 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     7321 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     7381 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     7441 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     7501 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     7561 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     7621 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     7681 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     7741 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     7801 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     7861 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     7921 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     7981 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     8041 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     8101 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     8161 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     8221 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     8281 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     8341 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     8401 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     8461 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     8521 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     8581 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     8641 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     8701 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     8761 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     8821 cgacggatcg ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat
     8881 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     8941 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     9001 cttagg
//
