LOCUS       V045812                 7732 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045812
VERSION     V045812
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             964..3222
                     /label="CNOT3(NM_014516)"
                     /note="CNOT3(NM_014516)"
                     /gene="CNOT3"
     CDS             3229..3294
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3332..3556
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3602..4030
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4044..4373
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4440..5234
                     /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    5408..5529
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5578..5594)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5602..5618
                     /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(5626..5656)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5671..5692
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5980..6565)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6736..7596)
                     /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(7597..7701)
                     /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 atccactagt ccagtgtggt ggaattcgcc
      961 accatggcgg acaagcgcaa actccaaggt gagattgatc gctgcctcaa gaaggtgtcc
     1021 gagggcgtgg agcagtttga agatatttgg cagaagctcc acaatgcagc caacgcgaac
     1081 cagaaagaaa agtatgaggc tgacctaaag aaggagatta agaagctaca acggctgagg
     1141 gaccaaatca agacatgggt agcgtccaac gagatcaagg acaagaggca gcttatagac
     1201 aaccgcaagc tcattgagac gcaaatggaa cggttcaaag ttgtggaacg agagaccaaa
     1261 accaaagctt acagcaaaga gggcctgggc ctggcccaga aggtagatcc tgcccagaag
     1321 gagaaggaag aggttggcca gtggctcacg aataccatcg acacgctcaa catgcaggtg
     1381 gaccagtttg agagtgaagt ggagtcactg tcagtgcaga cacgcaagaa gaagggcgac
     1441 aaggataagc aggaccggat tgagggcttg aagcggcaca tcgagaagca ccgctaccac
     1501 gtgcgcatgc tagagaccat cctgcgcatg ctggacaatg actccatcct cgttgacgcc
     1561 atccgcaaga tcaaggacga cgttgagtac tatgttgact catcccagga ccccgacttc
     1621 gaggagaacg agtttctcta cgatgacctg gacctcgagg acattccaca ggcgctggtc
     1681 gccacctccc cccccagcca cagccacatg gaggatgaga tcttcaacca gtccagcagc
     1741 acgcccacct caaccacctc cagctctccc atcccgccca gcccagccaa ctgtaccacg
     1801 gaaaactctg aagatgataa gaagagggga cgttccacag acagtgaagt cagccagtct
     1861 ccagccaaaa acggctccaa gcctgtccac agcaaccagc accctcagtc cccagctgtg
     1921 ccgcccacct acccctccgg ccccccgcct gctgcctctg ccttgagcac cactcctggc
     1981 aacaatgggg tccccgcccc cgcagcaccc ccaagtgccc tgggccccaa ggccagtcca
     2041 gctcccagcc acaactcggg cacccctgct ccctatgccc aggctgtggc cccaccagct
     2101 cccagtgggc ccagcacgac ccagccccgg ccccccagcg tccagcctag cggaggcgga
     2161 ggcggcggca gcggaggcgg agggagcagc agcagtagta acagcagtgc cggtggaggg
     2221 gctggcaagc agaatggcgc caccagttac agctcagttg tggcagacag cccggcagag
     2281 gtggctttga gcagcagtgg gggcaacaat gccagcagcc aggccttggg ccccccttcc
     2341 ggcccccaca acccacctcc cagcacctcg aaggaaccca gtgcggcagc cccaacgggg
     2401 gctgggggcg tggccccagg ctcagggaac aactcagggg gacccagcct cctggtgcca
     2461 ctgcctgtga atcctcccag ctccccaacg cccagcttca gtgatgccaa ggcagccggt
     2521 gccctgctca atgggcctcc acagttcagc accgccccag aaatcaaggc ccctgagcct
     2581 ctgagctcct tgaagtccat ggcggaacgg gcagccatca gctctggcat tgaggaccct
     2641 gtgccaacgc tgcacctgac cgagcgagac atcatcctga gcagtacatc agcacctccg
     2701 gcctcagccc agccgcccct gcagctgtca gaggtgaaca taccgctgtc gctgggtgtc
     2761 tgtccactgg gccctgtgcc cctcaccaag gagcagctct atcagcaggc catggaagag
     2821 gccgcctggc accacatgcc tcacccctct gactctgagc gtattcggca gtacctcccc
     2881 cggaacccct gtccgacgcc cccctaccac caccagatgc cacccccaca ctcggacact
     2941 gtggaattct accagcgcct gtcgaccgag acactcttct tcatcttcta ctatctggag
     3001 ggcactaagg cacagtatct ggcagccaag gccctaaaga agcagtcatg gcgattccac
     3061 accaagtaca tgatgtggtt ccagaggcac gaggagccca agaccatcac tgacgagttt
     3121 gagcagggca cctacatcta ctttgactac gagaagtggg gccagcggaa gaaggaaggc
     3181 ttcacctttg agtaccgcta cctggaggac cgggacctcc agctcgagga ctacaaagac
     3241 catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga caagtagtga
     3301 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     3361 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3421 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3481 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     3541 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     3601 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3661 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3721 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3781 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3841 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3901 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     3961 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     4021 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4081 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4141 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4201 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4261 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     4321 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     4381 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     4441 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     4501 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4561 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     4621 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4681 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4741 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     4801 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4861 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4921 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     4981 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     5041 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     5101 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     5161 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     5221 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     5281 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     5341 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5401 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5461 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5521 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     5581 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5641 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5701 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5761 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5821 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5881 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5941 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     6001 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6061 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6121 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     6181 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6241 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6301 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6361 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6421 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6481 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat
     6541 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6601 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6661 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6721 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6781 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6841 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6901 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6961 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7021 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7081 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7141 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7201 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7261 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7321 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     7381 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7441 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7501 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7561 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7621 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7681 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
