LOCUS       V024646                 7739 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024646
VERSION     V024646
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..3229
                     /label="DYRK1A(NM_001396)"
                     /note="DYRK1A(NM_001396)"
                     /gene="DYRK1A"
     CDS             3236..3301
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3339..3563
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3609..4037
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4051..4380
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4447..5241
                     /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    5415..5536
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5585..5601)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5609..5625
                     /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(5633..5663)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5678..5699
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5987..6572)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6743..7603)
                     /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(7604..7708)
                     /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 atgcatacag gaggagagac
      961 ttcagcatgc aaaccttcat ctgttcggct tgcaccgtca ttttcattcc atgctgctgg
     1021 ccttcagatg gctggacaga tgccccattc acatcagtac agtgaccgtc gccagccaaa
     1081 cataagtgac caacaggttt ctgccttatc atattctgac cagattcagc aacctctaac
     1141 taaccaggtg atgcctgata ttgtcatgtt acagaggcgg atgccccaaa ccttccgtga
     1201 cccagcaact gctcccctga gaaaactttc tgttgacttg atcaaaacat acaagcatat
     1261 taatgaggtt tactatgcaa aaaagaagcg aagacaccaa cagggccagg gagacgattc
     1321 tagtcataag aaggaacgga aggtttacaa tgatggttat gatgatgata actatgatta
     1381 tattgtaaaa aacggagaaa agtggatgga tcgttacgaa attgactcct tgataggcaa
     1441 aggttccttt ggacaggttg taaaggcata tgatcgtgtg gagcaagaat gggttgccat
     1501 taaaataata aagaacaaga aggcttttct gaatcaagca cagatagaag tgcgacttct
     1561 tgagctcatg aacaaacatg acactgaaat gaaatactac atagtgcatt tgaaacgcca
     1621 ctttatgttt cgaaaccatc tctgtttagt ttttgaaatg ctgtcctaca acctctatga
     1681 cttgctgaga aacaccaatt tccgaggggt ctctttgaac ctaacacgaa agtttgcgca
     1741 acagatgtgc actgcactgc ttttccttgc gactccagaa cttagtatca ttcactgtga
     1801 tctaaaacct gaaaatatcc ttctttgtaa ccccaaacgc agtgcaatca agatagttga
     1861 ctttggcagt tcttgtcagt tggggcagag gatataccag tatattcaga gtcgctttta
     1921 tcggtctcca gaggtgctac tgggaatgcc ttatgacctt gccattgata tgtggtccct
     1981 cgggtgtatt ttggttgaaa tgcacactgg agaacctctg ttcagtggtg ccaatgaggt
     2041 agatcagatg aataaaatag tggaagttct gggtattcca cctgctcata ttcttgacca
     2101 agcaccaaaa gcaagaaagt tctttgagaa gttgccagat ggcacttgga acttaaagaa
     2161 gaccaaagat ggaaaacggg agtacaaacc accaggaacc cgtaaacttc ataacattct
     2221 tggagtggaa acaggaggac ctggtgggcg acgtgctggg gagtcaggtc atacggtcgc
     2281 tgactacttg aagttcaaag acctcatttt aaggatgctt gattatgacc ccaaaactcg
     2341 aattcaacct tattatgctc tgcagcacag tttcttcaag aaaacagctg atgaaggtac
     2401 aaatacaagt aatagtgtat ctacaagccc cgccatggag cagtctcagt cttcgggcac
     2461 cacctccagt acatcgtcaa gctcaggtgg ctcatcgggg acaagcaaca gtgggagagc
     2521 ccggtcggat ccgacgcacc agcatcggca cagtggtggg cacttcacag ctgccgtgca
     2581 ggccatggac tgcgagacac acagtcccca ggtgcgtcag caatttcctg ctcctcttgg
     2641 ttggtcaggc actgaagctc ctacacaggt cactgttgaa actcatcctg ttcaagaaac
     2701 aacctttcat gtagcccctc aacagaatgc attgcatcat caccatggta acagttccca
     2761 tcaccatcac caccaccacc accatcacca ccaccatgga caacaagcct tgggtaaccg
     2821 gaccaggcca agggtctaca attctccaac gaatagctcc tctacccaag attctatgga
     2881 ggttggccac agtcaccact ccatgacatc cctgtcttcc tcaacgactt cttcctcgac
     2941 atcttcctcc tctactggta accaaggcaa tcaggcctac cagaatcgcc cagtggctgc
     3001 taataccttg gactttggac agaatggagc tatggacgtt aatttgaccg tctactccaa
     3061 tccccgccaa gagactggca tagctggaca tccaacatac caattttctg ctaatacagg
     3121 tcctgcacat tacatgactg aaggacatct gacaatgagg caaggggctg atagagaaga
     3181 gtcccccatg acaggagttt gtgtgcaaca gagtcctgta gctagctcgc tcgaggacta
     3241 caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa
     3301 gtagtgaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     3361 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     3421 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     3481 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     3541 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     3601 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3661 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3721 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3781 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3841 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3901 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3961 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     4021 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4081 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4141 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4201 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4261 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     4321 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4381 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     4441 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4501 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4561 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4621 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4681 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4741 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4801 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4861 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4921 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     4981 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     5041 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     5101 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     5161 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5221 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     5281 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     5341 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     5401 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5461 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5521 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     5581 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     5641 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     5701 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5761 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5821 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5881 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5941 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     6001 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     6061 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6121 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6181 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     6241 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6301 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     6361 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6421 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6481 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc
     6541 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6601 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6661 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6721 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6781 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6841 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6901 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6961 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7021 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     7081 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7141 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7201 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7261 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7321 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     7381 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7441 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7501 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     7561 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     7621 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     7681 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
