LOCUS       V019594                 7343 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019594
VERSION     V019594
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"
     CDS             683..2023
                     /label="CASP5(NM_001136112)"
                     /note="CASP5(NM_001136112)"
                     /gene="CASP5"
     CDS             2030..2095
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2119..2707
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2773..2821
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3023..3451
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3465..3794
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3861..4655
                     /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    4831..4952
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5001..5017)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5025..5041
                     /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(5049..5079)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5094..5115
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5403..5991)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6162..7022)
                     /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(7023..7127)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatggctgc tgtgcccaga gttgaaggag tctttatatt
      721 tctgatagaa gacagtggca aaaaaaaaag gcgtaagaat tttgaagcta tgttcaaagg
      781 tatccttcag agtggattgg ataacttcgt gataaaccac atgctaaaga acaacgtggc
      841 tggacaaaca tctatccaga ccctagtacc taatacggat caaaagtcga ccagtgtaaa
      901 aaaagacaac cacaaaaaaa aaacagttaa gatgttggaa tacctgggca aagatgttct
      961 tcatggtgtt tttaattatt tggcaaaaca cgatgttctg acattgaagg aagaggaaaa
     1021 gaaaaaatat tatgatacca aaattgaaga caaggccctg atcttggtag actctttgcg
     1081 aaagaatcgc gtggctcatc aaatgtttac ccaaacactt ctcaatatgg accaaaagat
     1141 caccagtgta aaacctcttc tgcaaatcga ggctggacca cctgagtcag cagaatctac
     1201 aaatatactc aaactttgtc ctcgtgaaga attcctgaga ctgtgtaaaa aaaatcatga
     1261 tgagatctat ccaataaaaa agagagagga ccgcagacgc ctggctctca tcatatgcaa
     1321 tacaaagttt gatcacctgc ctgcaaggaa tggggctcac tatgacatcg tggggatgaa
     1381 aaggctgctt caaggcctgg gctacactgt ggttgacgaa aagaatctca cagccaggga
     1441 tatggagtca gtgctgaggg catttgctgc cagaccagag cacaagtcct ctgacagcac
     1501 gttcttggta ctcatgtctc atggcatcct agagggaatc tgcggaactg cgcataaaaa
     1561 gaaaaaaccg gatgtgctgc tttatgacac catcttccag atattcaaca accgcaactg
     1621 cctcagtcta aaggacaaac ccaaggtcat cattgtccag gcctgcagag gtgaaaaaca
     1681 tggggaactc tgggtcagag actctccagc atccttggca ctcatctctt cacagtcatc
     1741 tgagaacctg gaggcagatt ctgtttgcaa gatccacgag gagaaggact tcattgcttt
     1801 ctgttcttca acaccacata acgtgtcctg gagagaccgc acaaggggct ccatcttcat
     1861 tacggaactc atcacatgct tccagaaata ttcttgctgc tgccacctaa tggaaatatt
     1921 tcggaaggta cagaaatcat ttgaagttcc acaggctaaa gcccagatgc ccaccataga
     1981 acgagcaacc ttgacaagag atttctacct ctttcctggc aatctcgagg actacaaaga
     2041 ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg acaagtagtg
     2101 agggcccgat atctcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     2161 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     2221 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     2281 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     2341 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     2401 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     2461 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg
     2521 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     2581 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     2641 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     2701 tccccgcctg gaaacggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     2761 cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt
     2821 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     2881 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     2941 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc
     3001 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3061 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3121 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggc
     3181 atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3241 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3301 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3361 tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg gttaaaaaat
     3421 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     3481 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3541 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3601 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3661 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3721 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3781 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     3841 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3901 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3961 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4021 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     4081 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4141 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4201 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4261 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4321 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4381 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4441 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4501 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4561 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4621 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgcgaaatg
     4681 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     4741 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4801 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     4861 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4921 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     4981 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     5041 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     5101 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     5161 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5221 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     5281 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     5341 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5401 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5461 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5521 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5581 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5641 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5701 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5761 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5821 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     5881 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5941 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6001 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6061 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     6121 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6181 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6241 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6301 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6361 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6421 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6481 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6541 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6601 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6661 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6721 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     6781 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6841 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6901 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     6961 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7021 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7081 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     7141 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tcgactctca
     7201 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg
     7261 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca
     7321 attgcatgaa gaatctgctt agg
//
