LOCUS       V027716                 8369 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027716
VERSION     V027716
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..3859
                     /label="Nlrx1(NM_001025010)"
                     /note="Nlrx1(NM_001025010)"
                     /gene="Nlrx1"
     CDS             3866..3931
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3969..4193
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4239..4667
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4681..5010
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5077..5871
                     /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    6045..6166
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6215..6231)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6239..6255
                     /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(6263..6293)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6308..6329
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6617..7202)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7373..8233)
                     /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(8234..8338)
                     /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 atgaggtggg gctgccattt
      961 gcccaggacc tcttggggct ctggcctggg aagaacagtc ccgctcccag acgagctccg
     1021 gattcagtcg agctggccct ttaaaggggt gcatcatccc cttaggcccc ctagggcctt
     1081 tatccgtcac catggaaact cggcagacag tgctccccca ccagggaggc atgggcagct
     1141 gttcaggagc atctctgcca ctgaagctat ccaaaggcat cgtcggaacc tcactgagtg
     1201 gtttagccgg ctgcccagag aggagcgcca gtttggacca acctttgctc tagacacagt
     1261 tcacgttgac cccgtgatcc gagagagcac cccagatgac ctgcttcgtc catccacgga
     1321 gctggccaca gggcatcagc gaacccaggc agagctcccc ccactggccc tgtctcagct
     1381 ctttgaccca gatgcctgtg ggcgccgcgt gcagacagtg gtattgtatg ggaccgtggg
     1441 tactggcaag agcacattgg tgcgcaagat ggtcttagac tggtgttatg ggagactgcc
     1501 tgcctttgag ctgctcatcc ccttctcctg tgaggacctg tcatccctgg gctccacccc
     1561 agcttcctta tgccaacttg tgacccagcg ttacacaccc ctgaaagagg tgttgcccct
     1621 gatgaatgct gctggatccc gcctgctctt tgtgctccat ggcttggaac gcctcaacct
     1681 tgacttccgg ctggcaggca cagggctttg cagtgacccg gaggaacccg gggcaccagc
     1741 tgccatcatg gtcaacctgc tgcgcaaata catgcttcct gaggccagca ttctggtaac
     1801 cacccggccc tccgccattg gccgtatccc tagcaagtac gtgggccgct atggtgaaat
     1861 ctgtggcttc tcggatacca acctgcagaa gctctacttc cagctccgcc ttaaccagcc
     1921 tgactgtggg tatggtgctg ggggtacagg tgtctcagtt acaccagctc agcgggacaa
     1981 tctcattcaa atgctgtccc gaaacctgga ggggcaccac cagatcgcag ccgcctgctt
     2041 tctgccctcc tactgctggc ttgtctgtgc taccttgcac ttcctgcatg ctcccacatc
     2101 tgctggtcag actctcacaa gcatctatac cagctttcta cgtctgaact tcaatgggga
     2161 aacactggac agcaccgacc cctccaactt atccctgatg tcctatgcag cccggactat
     2221 gggtaagttg gcctatgagg gggtgtcatc ccgaaagacc tacttctctg aagaggatgt
     2281 ccgtggctgc ctggaagctg gcatcaagac agaggaggag tttcagctgc tccagatctt
     2341 ccgccgggat gccctgaggt ttttcctggc cccgtgtgtg gaaccagggc acctgggtac
     2401 cttcgtattc accgtacctg ccatgcagga gtatctggct gccctctaca tcgtgcttgg
     2461 tttgcgcaag acagccctgc agcgggtggg caaagaagtg tttgaatttg tgggccgtgt
     2521 tggggaagat gtcagcctgg tattgggcat tgtggccaaa ctgttgcccc tgcggattct
     2581 gcctctgcta ttcaacttgc ttaaggtggt tccacgagtg tttgggcgca tggtgagtaa
     2641 gagtcgcgag gcggtggctc aggccatggt gctggagatg ttccgggagg aagactatta
     2701 caatgacgat gttctggatc agatgggtgc cagcatcctg ggtgtggagg gcccccggcg
     2761 ccacccagat gaaccccctg aggatgaagt ctttgagctc ttccccatgt tcatgagtgg
     2821 acttctctct gcccacaatc gggcggtgtt ggctcagctt ggctgtccca ttaagaacct
     2881 ggatgccctg gagaatgccc aggcaatcaa gaagaaactg gggaagatgg gtcggcaggt
     2941 gctgcccccc tcggagcttc ttgaccatct cttctttcac tatgagttcc agaaccagcg
     3001 cttctcagct gaggtgctgg gctccctacg acagctcaat ttagcaggtg tgcgaatgac
     3061 acccctcaag tgcacggtgg tagcgtctgt actgggaagt ggacggcacc ccctggatga
     3121 agtgaacttg gcctcctgcc agctggaccc cgctgggcta cacactctca tgcctgttct
     3181 cctgcgggcc cggaaactgg ggttgcaact caacaatctg ggccctgagg cctgcagaga
     3241 cctccgagac ctgctgttac atgaccaatg ccagatcacc actctccggc tgtccaacaa
     3301 cccactgaca gcagcagggg tgggtgtgct gatggacggg ctggcaggaa acacttcggt
     3361 gacacacctg tccctgctgc acactgacct tggagatgag ggactggagt tgctggctgc
     3421 ccagctggac cgtaacaaac aactacagga actgaatgtg gcctacaatg gtgctgggga
     3481 cacggtggct ctggccttgg ctaaggctgc tcggaagcac ccgtccctgg agctgctgca
     3541 cctctacttc aatgagctga gttcagaggg tcgccaggtc ctgcgggatt tgggcggctc
     3601 tggcgaaggt ggtgcccggg ttgtagcctc gctgacagag gggacagcgg tgtctgagta
     3661 ctggtcagtg atcctcagtg aagtccaacg caacctcaac agctgggacc ctgtccgggt
     3721 ccagcgccat ctcaagctgc tgctccgaga cttggaagac agcagaggtg ccacccttaa
     3781 tccttggcgc aaggctcagc ttctgcgggt ggagagcgag gtcaagactc ttctggagca
     3841 gctgggaggc tctggacacc tcgaggacta caaagaccat gacggtgatt ataaagatca
     3901 tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgtttaaa cccgctgatc
     3961 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     4021 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     4081 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     4141 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     4201 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     4261 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     4321 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4381 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4441 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4501 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4561 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     4621 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     4681 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4741 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     4801 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4861 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4921 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     4981 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     5041 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     5101 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     5161 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     5221 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     5281 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     5341 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     5401 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     5461 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     5521 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     5581 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     5641 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5701 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5761 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     5821 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5881 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     5941 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     6001 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     6061 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     6121 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     6181 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     6241 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     6301 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     6361 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     6421 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     6481 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6541 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6601 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6661 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6721 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6781 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6841 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6901 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6961 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     7021 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     7081 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     7141 ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7201 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7261 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7321 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7381 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7441 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7501 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7561 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7621 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7681 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7741 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7801 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7861 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7921 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7981 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     8041 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     8101 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     8161 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     8221 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8281 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8341 catttccccg aaaagtgcca cctgacgtc
//
