LOCUS       Exported                7196 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses mouse DYRK4 tagged with 2A peptide at it's C-terminus and 
            GFP in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-DYRK4-2A-GFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7196)
  AUTHORS   Slepak TI, Salay LD, Lemmon VP, Bixby JL
  TITLE     Dyrk kinases regulate phosphorylation of doublecortin, cytoskeletal 
            organization, and neuronal morphology.
  JOURNAL   Cytoskeleton (Hoboken). 2012 Jul;69(7):514-27. doi: 
            10.1002/cm.21021. Epub 2012 Mar 7.
  PUBMED    22359282
REFERENCE   2  (bases 1 to 7196)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7196
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     enhancer        221..524
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        525..727
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     678..698
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     724..748
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2926..2937
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             3131..3184
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3212..3928
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             3212..3925
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(3254..3275)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3515..3534)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3862..3883
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3943..4419
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4142..4161)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4459..4599
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4459..4588
                     /label=AAV2 ITR
     rep_origin      4674..5129
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4761..4780)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4971..4992
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5146..5165)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5265..5287
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5325..5343)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5411..5515
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5516..6376
                     /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"
     primer_bind     complement(5734..5753)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6547..7135
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7036..7055
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtggagc tagttattaa tagtaatcaa
      181 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
      241 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      301 ttcccatagt aacgtcaata gggactttcc attgacgtca atgggtggag tatttacggt
      361 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      421 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      481 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
      541 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
      601 ttgacgtcaa tgggagtttg ttttgcacca aaatcaacgg gactttccaa aatgtcgtaa
      661 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      721 cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct
      781 ccatagaaga caccgggacc gatccagcct ccgcggattc gaatcccggc cgggaacggt
      841 gcattggaac gcggattccc cgtgccaaga gtgacgtaag taccgcctat agagtctata
      901 ggcccacaaa aaatgctttc ttcttttaat atactttttt gtttatctta tttctaatac
      961 tttccctaat ctctttcttt cagggcaata atgatacaat gtatcatgcc tctttgcacc
     1021 attctaaaga ataacagtga taatttctgg gttaaggcaa tagcaatatt tctgcatata
     1081 aatatttctg catataaatt gtaactgatg taagaggttt catattgcta atagcagcta
     1141 caatccagct accattctgc ttttatttta tggttgggat aaggctggat tattctgagt
     1201 ccaagctagg cccttttgct aatcatgttc atacctctta tcttcctccc acagctcctg
     1261 ggcaacgtgc tggtctgtgt gctggcccat cactttggca aagaattggg attcgaacat
     1321 cgattgaatt ccccggggat ccatgaggac gtccaaggct cagtcggagg ttgagagtaa
     1381 gcctttgggt cacctccaga agccatcctc caagaacaag aagctgagag tgaaccgatg
     1441 tccacaaaag attccaagta acacggcgtt cccttttgtg gacacaaagg ggaagaggaa
     1501 cacagttaac ttcccccaga ttggcaacaa agtcccatca aaggggccca tgcagtatca
     1561 ggaaaatcaa attcataatc aagtgctatc ctctgaactc aagacttcag aaataccttt
     1621 caacattaac accaaagccc aagacaccaa gccacaccca gaactacaaa agaagcacaa
     1681 agtccctctt acagtagcag aggccctaaa gttctttaag aatcagctgt ccccttatga
     1741 gcaaagcgaa atcctgggct actcagagct gtggttcctg ggccttgaag ccaagaagct
     1801 caacgtggtt ccagagaagt tcagcaagac aagctttgat gatgagcacg gttcctatat
     1861 gaaggtcctg catgaccaca ttgcctaccg atatgaggtt ctggagatga tcgggaaagg
     1921 gtcctttgga caggtggcca agtgcttgga tcacaaaaac aatgagttgg tggccctgaa
     1981 aatcatcagg aacaaaaaga gatttcacca tcaggctctg gtggagctga agatcctgga
     2041 agccctcaga aggaaggaca aagacaacaa ccacaatgtg gtccacatga aggatttctt
     2101 ctacttccgg aatcacctct gtatcacctt tgagctcctg ggaatcaacc tgtatgagtt
     2161 gatgaagaac aacagctttc atggcttcaa tctgtccata gttcggcgct tcaccttttc
     2221 cattttgaaa tgcttacaca tgctttatgt agagaaaatc atccactgtg acctcaagcc
     2281 ggaaaatatt gtgctatacc agaggggcca agtcactgtt aaagttattg actttgggtc
     2341 aagctgttat gaacaccaga aagtgtatac gtacatccaa agtcgtttct accgatcccc
     2401 agaggtgatt ctgggacacc cctataacat ggccattgac atgtggagcc tgggctgcat
     2461 catggcggag ctgtacaccg gatacccact gttccctggg gagaacgagg tggagcagct
     2521 ggcttgcatc atggaggtgc tgggcctgcc acctgcccac ttcacccaga cagcctccag
     2581 gagacaggta ttctttgatt ccaaaggtct tcccaaaaat ataaacaaca acagaggggg
     2641 aaaaagatac ccagattcca aggatctcac gatggtggtg aaaacctacg attccagctt
     2701 cctggacttt ctcagaaggt gggagccttc tctgcgcatg acccccgagc aggccctcaa
     2761 gcatgcctgg attcacgagc cgaggaaatt caaaccacgg cccaagccac agatattaag
     2821 gaagcctggt gccagtattt cctctgagat aagcacggaa aaggctgaag agcaacaggc
     2881 cagcaaaggc aaaaaagatg aggctaccaa agaaactaca gacaaattga aggacgaagc
     2941 tgagaagcac cttgagaatt cgggcaaaca gcagagctct gtggagcaca cagcagacac
     3001 catccagctg cctcacctaa cagaagcttc tggaaagtca gagactgttg ctgggtcaga
     3061 gatgtccgca gaacagcaga gcacctcctc ccccaagagc acaaacatct taccacccat
     3121 cgtgaccggt gaaggccgcg gcagcctgct gacctgcggc gatgtggaag aaaacccggg
     3181 cccggcgccg ggcagcacta gtcggaagct tgtgagcaag ggcgaggagc tgttcaccgg
     3241 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     3301 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     3361 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
     3421 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     3481 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     3541 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     3601 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     3661 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
     3721 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     3781 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga
     3841 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     3901 tctcggcatg gacgagctgt acaagtaact cgagagatct acgggtggca tccctgtgac
     3961 ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc
     4021 ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg
     4081 tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg
     4141 gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc
     4201 tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc
     4261 atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc
     4321 aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg
     4381 ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg
     4441 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     4501 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     4561 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc
     4621 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     4681 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4741 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4801 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4861 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     4921 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4981 gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat
     5041 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     5101 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     5161 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     5221 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     5281 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     5341 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     5401 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     5461 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     5521 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     5581 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     5641 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5701 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     5761 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5821 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5881 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5941 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     6001 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     6061 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     6121 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     6181 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     6241 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     6301 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     6361 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     6421 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6481 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6541 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6601 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6661 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     6721 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6781 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6841 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6901 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6961 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     7021 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     7081 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     7141 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
