LOCUS       V028479                 9596 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028479
VERSION     V028479
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..5086
                     /label="NINL(NM_025176)"
                     /note="NINL(NM_025176)"
                     /gene="NINL"
     CDS             5093..5158
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    5196..5420
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5466..5894
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5908..6237
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6304..7098
                     /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    7272..7393
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7442..7458)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7466..7482
                     /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(7490..7520)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7535..7556
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7844..8429)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8600..9460)
                     /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(9461..9565)
                     /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 atggatgaag aagagaacca
      961 ctatgtctcg cagctcaggg aagtctacag cagctgcgac accacgggga ctggctttct
     1021 ggaccgccag gagctgaccc agctctgcct taagcttcac ctggagcagc agctgcccgt
     1081 cctcctgcag acgcttctcg gaaacgacca tttcgccagg gttaactttg aggaatttaa
     1141 ggaaggtttt gtggctgtgt tgtcttcaaa tgctggtgtt cgcccctcag atgaagacag
     1201 tagttctttg gaatcagctg cctccagtgc catccctcca aagtatgtga atggttctaa
     1261 gtggtatggc cgtcggagcc ggcctgagct atgtgacgct gccacagaag ccagacgcgt
     1321 gccggagcag caaacccagg ccagcctgaa aagtcacctc tggcgctcag cgtctctgga
     1381 gagcgtggag agtcccaagt cagatgaaga ggccgagagc actaaagaag ctcagaatga
     1441 attatttgaa gcacaaggac agctgcagac ctgggattct gaggactttg ggagccccca
     1501 gaagtcctgc agcccctcct ttgacacccc agagagccag atccggggcg tgtgggaaga
     1561 gctgggggtg ggcagcagcg gacacctgag cgagcaggag ctggctgtgg tctgccagag
     1621 cgtcgggctc cagggactcg agaaagagga actcgaagac ctgtttaaca aactggatca
     1681 agacggagac ggcaaagtga gtcttgagga attccagctt ggcctcttca gtcatgagcc
     1741 cgcgctactt ctagagtctt ccactcgggt taaaccgagc aaggcttggt ctcattacca
     1801 ggtcccagag gagagcggct gccacaccac cacaacctca tccctcgtgt ccctgtgctc
     1861 cagcctgcgc ctcttctcca gcattgacga tggttctggc ttcgcttttc ctgatcaggt
     1921 cctggccatg tggacccagg aggggattca gaatggcagg gagatcttgc agagcctgga
     1981 cttcagcgtg gacgagaagg tgaaccttct ggagctgacc tgggcccttg acaacgagct
     2041 catgacagtg gacagtgccg tccagcaggc agccctggcc tgctaccacc aggagctgag
     2101 ctaccagcaa gggcaggtgg agcagctggc aagggagcgt gacaaggcaa ggcaggacct
     2161 ggagagggcc gagaagagga acctggagtt tgtgaaagag atggacgact gccactccac
     2221 cctggagcag ctcacggaga agaaaatcaa gcatctggag caggggtacc gggaaaggct
     2281 gagcctcctg cggtctgagg tggaggcgga gcgagagctg ttctgggagc aggcccacag
     2341 gcagagggcc gcgctggagt gggacgtggg gcgcctgcag gctgaggagg ctggcctccg
     2401 cgagaagctg accctggccc tgaaggaaaa cagtcgccta cagaaggaga ttgtggaagt
     2461 ggtggaaaag ctttcggatt cggagaggct ggccctgaag ctgcagaagg acctggagtt
     2521 tgtgctgaag gacaagctgg agccacagag tgcagagctc ctggcccagg aggagcggtt
     2581 cgcagcagtc ctgaaggaat acgagctcaa gtgccgggac ctgcaggacc gcaacgatga
     2641 gctgcaagct gagctggaag gcctgtgggc gcggctgccc aagaaccggc acagcccctc
     2701 atggagcccg gatgggcgca gacggcagct ccctggactc ggcccagcag gcatttcatt
     2761 cctgggtaat tctgctccag tgagtataga aacggagctg atgatggagc aggtaaagga
     2821 gcattaccaa gacctcagga cccagctgga gaccaaggta aattactacg aaagggaaat
     2881 tgcggcactg aaaaggaact ttgagaagga gaggaaggac atggagcagg ctcgcaggcg
     2941 cgaggtcagc gtgctggagg gtcagaaggc cgacctggag gagctccacg agaagtctca
     3001 ggaggtcatc tggggcctgc aggagcagct gcaggacaca gcccgcggcc ccgagcctga
     3061 gcagatgggc ctggcaccct gctgcaccca ggcactgtgt ggcctggccc tgcggcatca
     3121 cagccacctg cagcagatca ggagagaggc tgaggcggag ctgagtggag agctgtcggg
     3181 gctgggagcc ctgcccgctc gcagagacct gaccttggag ctggaggagc cgccgcaggg
     3241 acccctgcca cgcgggagcc agaggtcgga gcagctggag ctggagaggg cactgaagct
     3301 gcagccctgt gcgagcgaga agcgcgccca gatgtgcgta tcgttggccc tcgaggagga
     3361 ggagttggag cttgcccgcg ggaagcgagt ggacgggccc tccctggaag cagagatgca
     3421 ggccctgccg aaagatgggc tggtggcagg aagtggccag gagggcacac gtggcctcct
     3481 accactgcgt ccgggctgtg gggagcggcc actggcctgg ctggccccag gtgatggcag
     3541 agagtctgag gaggcggcag gagccgggcc tcgccgcagg caagcccagg acacagaagc
     3601 tacgcagagc ccggcccccg cccctgcccc ggcatcccac ggcccctcag agaggtggtc
     3661 acgcatgcag ccctgtggag tggatgggga tattgtccca aaggagccag agcctttcgg
     3721 cgcgagcgca gcggggctgg agcagcctgg agcccgggag ctgcctctgc tgggaacaga
     3781 gagagacgcc tcgcaaaccc agccacggat gtgggagcca cccctgaggc cggccgcttc
     3841 gtgcagggga caggctgaga ggctacaggc cattcaggaa gagcgagcac gaagctggag
     3901 caggggcacc caggagcagg cctcggagca gcaggcccgg gccgagggcg ccctggagcc
     3961 tgggtgtcac aagcacagtg tggaggttgc caggagaggg tccttgccat cccacctcca
     4021 gctcgcagac ccgcagggtt cctggcagga gcagcttgct gccccagaag agggggagac
     4081 caaaatagcg ctggagagag agaaggatga catggaaacc aaacttctac atctggaaga
     4141 cgtcgtccgg gctctggaga aacatgtaga tttgagagag aacgacagac tggagttcca
     4201 tagactttct gaagaaaaca ctttgttgaa aaacgatctg ggaagggttc ggcaagagct
     4261 tgaagctgca gaaagtactc acgatgcaca gaggaaggaa attgaggttt taaagaaaga
     4321 caaggaaaag gcctgctctg agatggaggt gctcaacaga cagaatcaga actacaagga
     4381 tcaattatcc cagctcaatg tcagggttct tcaactggga caggaggctt ctacccacca
     4441 ggcccaaaac gaggagcatc gtgtgaccat tcagatgtta acacagagcc tggaggaggt
     4501 ggttcgcagt gggcagcagc agagtgacca aatccaaaaa cttagagttg aacttgaatg
     4561 cctgaatcag gaacatcaga gcctgcagct gccatggtca gagctgaccc agacccttga
     4621 ggaaagtcaa gaccaggtgc agggagctca cctgaggctg aggcaggccc aggcccagca
     4681 cttgcaggag gtccggctgg tgccccagga ccgtgtggcc gagctgcatc gcctgctcag
     4741 ccttcaggga gagcaggcca ggaggcgcct ggatgcacag cgggaagaac atgagaaaca
     4801 gctgaaagcc acagaagagc gggtggaaga ggcggagatg attctgaaga atatggaaat
     4861 gctcctccaa gagaaagtgg ataagctgaa ggagcagttt gaaaagaaca cgaagtccga
     4921 cctgctgctg aaggagctgt acgtggagaa cgcccacctg gtgagagcac ttcaggccac
     4981 cgaggagaag cagcgaggcg ccgagaaaca aagccgcctc ttggaagaaa aagttcgcgc
     5041 tctcaacaaa ctcgtcagta ggattgcccc cgcagccctc tctgtgctcg aggactacaa
     5101 agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta
     5161 gtgagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     5221 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     5281 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     5341 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     5401 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     5461 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     5521 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     5581 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     5641 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     5701 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     5761 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     5821 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     5881 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     5941 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     6001 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     6061 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     6121 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     6181 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     6241 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     6301 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     6361 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     6421 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     6481 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     6541 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     6601 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     6661 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     6721 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     6781 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     6841 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     6901 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     6961 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     7021 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     7081 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     7141 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     7201 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     7261 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     7321 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     7381 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     7441 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     7501 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     7561 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     7621 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     7681 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     7741 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     7801 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     7861 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     7921 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     7981 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     8041 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     8101 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     8161 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     8221 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     8281 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     8341 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc
     8401 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     8461 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     8521 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     8581 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     8641 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     8701 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     8761 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     8821 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     8881 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     8941 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     9001 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     9061 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     9121 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     9181 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     9241 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     9301 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     9361 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     9421 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     9481 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     9541 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
