LOCUS       Exported                7336 bp DNA     circular SYN 07-JUN-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7336)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 22, 2021 from SnapGene Viewer 5.3.1
            https://www.snapgene.com
COMMENT     in-silico sequence added (B1 - Insert your ORF - B1)
            
            #################
            open pCMV 3xFLAG 7.1 from Sigma with EcoRV put in rfB = pDEST 3xFLAG
            cmv 7.1 AmpR
            
            3xFLAG_s   ccgccccgttgacgcaaatggg pos 840-861
            3xFLAG_as  acccgggatcctctagagtcga  pos 2753-2774
            
            ##################
            Note: GATaTC, EcoRV site at B2 site is mutated to GATTC
FEATURES             Location/Qualifiers
     source          1..7336
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        166..916
                     /label=CMV promotor
                     /note="CMV promotor"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     840..861
                     /label=Forward sequencing primer
                     /note="Forward sequencing primer"
     CDS             931..996
                     /codon_start=1
                     /label=3xFLAG
                     /note="3xFLAG"
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    1033..1057
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1060..3618
                     /codon_start=1
                     /label=TAOK1 var2 Nop
                     /note="isoform 2 is encoded by transcript variant 2; 
                     serine/threonine protein kinase TAO1 homolog; microtubule 
                     affinity regulating kinase kinase; serine/threonine-protein
                     kinase TAO1; STE20-like kinase PSK2; kinase from chicken 
                     homolog B; thousand and one amino acid protein 1; MARK 
                     Kinase; prostate-derived STE20-like kinase 2; 
                     prostate-derived sterile 20-like kinase 2; thousand and one
                     amino acid protein kinase 1"
                     /translation="PSTNRAGSLKDPEIAELFFKEDPEKLFTDLREIGHGSFGAVYFAR
                     DVRTNEVVAIKKMSYSGKQSTEKWQDIIKEVKFLQRIKHPNSIEYKGCYLREHTAWLVM
                     EYCLGSASDLLEVHKKPLQEVEIAAITHGALQGLAYLHSHTMIHRDIKAGNILLTEPGQ
                     VKLADFGSASMASPANSFVGTPYWMAPEVILAMDEGQYDGKVDVWSLGITCIELAERKP
                     PLFNMNAMSALYHIAQNESPTLQSNEWSDYFRNFVDSCLQKIPQDRPTSEELLKHIFVL
                     RERPETVLIDLIQRTKDAVRELDNLQYRKMKKLLFQEAHNGPAVEAQEEEEEQDHGVGR
                     TGTVNSVGSNQSIPSMSISASSQSSSVNSLPDVSDDKSELDMMEGDHTVMSNSSVIHLK
                     PEEENYREEGDPRTRASDPQSPPQVSRHKSHYRNREHFATIRTASLVTRQMQEHEQDSE
                     LREQMSGYKRMRRQHQKQLMTLENKLKAEMDEHRLRLDKDLETQRNNFAAEMEKLIKKH
                     QAAMEKEAKVMSNEEKKFQQHIQAQQKKELNSFLESQKREYKLRKEQLKESKELQIKKQ
                     FQDTCKIQTRQYKALRNHLLETTPKSEHKAVLKRLKEEQTRKLAILAEQYDHSINEMLS
                     TQALRLDEAQEAECQVLKMQLQQELELLNAYQSKIKMQAEAQHDRELRELEQRVSLRRA
                     LLEQKIEEEMLALQNERTERIRSLLERQAREIEAFDSESMRLGFSNMVLSNLSPEAFSH
                     SYPGASGWSHNPTGGPGPHWGHPMGGPPQAWGHPMQGGPQPWGHPSGPMQGVPRGSSMG
                     VRNSPQALRRTASGGRTEQGMSRSTSVTSQISNGSHMSYT"
     protein_bind    complement(3620..3644)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     3659..3680
                     /label=Reverse sequencing primer
                     /note="Reverse sequencing primer"
     polyA_signal    3677..4299
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     promoter        4328..4657
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4508..4643
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(4696..4714)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4728..4744)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4752..4768
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(4776..4806)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4821..4842
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5115..5734)
                     /direction=LEFT
                     /label=pBR322 ori
                     /note="pBR322 ori"
     rep_origin      complement(5130..5718)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5889..6749)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6750..6854)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6791..6819
                     /label=E7 amp promotor
                     /note="E7 amp promotor"
     rep_origin      6881..7336
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa
      961 gatcatgaca tcgattacaa ggatgacgat gacaagcttg cggccgcgaa ttcatcgata
     1021 gatctgatat caacaagttt gtacaaaaaa gcaggctccc catcaactaa cagagcaggc
     1081 agcctgaagg accctgaaat tgcagagctc ttcttcaaag aagatccaga gaagctcttc
     1141 acagatctca gagaaattgg ccatggaagc tttggagcag tgtattttgc acgagatgtg
     1201 cgtaccaatg aagtggtggc catcaagaaa atgtcttata gtggaaagca gtctactgag
     1261 aaatggcagg atattattaa ggaagtcaag tttctacaaa gaataaaaca tcccaacagt
     1321 atagaataca aaggctgtta tttacgtgaa cacacagcat ggcttgtaat ggaatattgt
     1381 ttaggatctg cttcggattt actagaagtt cacaaaaagc cattacaaga agtggaaata
     1441 gcagcaatta cacatggtgc tcttcaggga ttagcctact tacattctca tactatgatt
     1501 catagagata tcaaagcagg aaatatcctt ctgacagaac caggccaggt gaaacttgct
     1561 gactttggct ctgcttccat ggcatcacct gccaattcct ttgtgggaac gccgtattgg
     1621 atggccccag aagtaatttt agccatggat gaaggacaat atgatggcaa agtagatgtg
     1681 tggtctcttg gaataacatg tattgaacta gcggaaagga agcctccttt atttaatatg
     1741 aatgcaatga gtgccttata tcacatagcc caaaatgaat cccctacact acagtctaat
     1801 gaatggtctg attattttcg caactttgta gattcttgcc tccagaaaat ccctcaagat
     1861 cgacctacat cagaggaact tttaaagcac atatttgttc ttcgggagcg ccctgaaacc
     1921 gtgttaatag atctcattca gaggacaaag gatgcagtaa gagagctgga caatctgcag
     1981 tatcgaaaga tgaagaaact ccttttccag gaggcacata atggaccagc agtagaagca
     2041 caggaagaag aagaggaaca agatcatggt gttggccgga caggaacagt taatagtgtt
     2101 ggaagtaatc aatccattcc cagcatgtcc atcagtgcca gcagccaaag cagtagtgtt
     2161 aacagtcttc cagatgtctc agatgacaag agtgagctag acatgatgga gggagaccac
     2221 acagtgatgt ctaacagttc tgttatccat ttaaaaccag aggaagaaaa ttacagagaa
     2281 gagggagatc ctagaacaag agcatcagat ccacaatctc caccccaagt atctcgtcac
     2341 aaatcacact atcgtaatcg agaacacttt gctactatac ggacagcatc actggttacg
     2401 aggcaaatgc aagaacatga gcaggactct gagcttagag aacaaatgtc tggctataag
     2461 cgaatgaggc gacaacatca aaagcaactg atgactctgg aaaacaagct aaaggctgag
     2521 atggatgaac atcgcctcag attagacaaa gatcttgaaa ctcagcgtaa caattttgct
     2581 gcagaaatgg agaaacttat caagaaacac caggctgcca tggagaaaga ggctaaagtg
     2641 atgtccaatg aagagaaaaa atttcagcaa catattcagg cccaacagaa gaaagaactg
     2701 aatagttttc tcgagtccca gaaaagagag tataaacttc gaaaagagca gcttaaagag
     2761 tctaaagaac tccaaataaa aaagcagttt caggatacct gcaaaatcca aaccagacag
     2821 tacaaagcat taagaaatca cctgctggag actacaccaa agagtgagca caaagctgtt
     2881 ctgaaacggc tcaaggagga acagacccgg aaattagcta tcttggctga gcagtatgat
     2941 cacagcatta atgaaatgct ctccacacaa gccctgcgtt tggatgaagc acaggaagca
     3001 gagtgccagg ttttgaagat gcagctgcag caggaactgg agctgttgaa tgcgtatcag
     3061 agcaaaatca agatgcaagc tgaggcacaa catgatcgag agcttcgcga gcttgaacag
     3121 agggtctccc tccggagggc actcttagaa caaaagattg aagaagagat gttggctttg
     3181 cagaatgagc gcacagaacg aatacgaagc ctgttggaac gtcaagccag agagattgaa
     3241 gcttttgact ctgaaagcat gagactaggt tttagtaata tggtcctttc taatctctcc
     3301 cctgaggcat tcagccacag ctacccggga gcttctggtt ggtcacacaa ccctactggg
     3361 ggtccaggac ctcactgggg tcatcccatg ggtggcccac cacaagcttg gggccatcca
     3421 atgcaaggtg gaccccagcc atggggtcac ccttcagggc caatgcaagg ggtacctcga
     3481 ggtagcagta tgggagtccg caatagcccc caggctctga ggcggacagc ttctggggga
     3541 cggacggagc agggcatgag cagaagcacg agtgtcactt cacaaatatc caatgggtca
     3601 cacatgtctt atacataaga cccagctttc ttgtacaaag tggttgattc ggtaccagtc
     3661 gactctagag gatcccgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct
     3721 ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt
     3781 gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg
     3841 gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc
     3901 agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc
     3961 tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt
     4021 tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca
     4081 ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc
     4141 cttccctgtc cttctgattt taaaataact ataccagcag gaggacgtcc agacacagca
     4201 taggctacct ggccatgccc aaccggtggg acatttgagt tgcttgcttg gcactgtcct
     4261 ctcatgcgtt gggtccactc agtagatgcc tgttgaattg ggtacgcggc cagcttggct
     4321 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4381 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     4441 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     4501 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     4561 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     4621 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcctcgagg aactgaaaaa
     4681 ccagaaagtt aattccctat agtgagtcgt attaaattcg taatcatgtc atagctgttt
     4741 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     4801 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     4861 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     4921 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4981 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5041 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5101 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5161 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5221 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5281 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5341 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5401 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5461 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5521 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     5581 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5641 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5701 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5761 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5821 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5881 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5941 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6001 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6061 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6121 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6181 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6241 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6301 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6361 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6421 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     6481 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6541 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6601 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6661 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6721 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6781 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6841 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca
     6901 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     6961 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     7021 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     7081 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     7141 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     7201 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     7261 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     7321 ttaacgctta caattt
//
