LOCUS       sequence-313748-        6366 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6366)
  AUTHORS   Shen K, Meyer T
  TITLE     Dynamic control of CaMKII translocation and localization in 
            hippocampal neurons by NMDA receptor stimulation.
  JOURNAL   Science. 1999 Apr 2. 284(5411):162-6.
  PUBMED    10102820
REFERENCE   2  (bases 1 to 6366)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science. 
            1999 Apr 2. 284(5411):162-6."
COMMENT     Sequence Label: pEGFP-C1-CaMKIIbeta(K43R)
FEATURES             Location/Qualifiers
     source          1..6366
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(553..555,556..558,559..1269)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(553..555,556..558,559..1269)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(598..619)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(859..878)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1206..1227
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1270..1335
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    3094..3215
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3131..3150)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3185..3204
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3222..3677)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3359..3380)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3571..3590
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3704..3808
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3810..4167
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3810..4001
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3833..3853)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4018..4153
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4202..4996
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4202..4996
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4256..4275)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4866..4885
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5184..5203)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5228..5275
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5604..6192
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6093..6112
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      601 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      661 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      721 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      781 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      841 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      901 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      961 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1021 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1081 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1141 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1201 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1261 ctgtacaagt ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc
     1321 gcgggcccgg gatccaccgg atctagaatg gccaccacgg tgacctgcac ccgtttcacg
     1381 gacgagtacc agctatacga ggatattggc aagggggctt tctctgtggt ccgacgctgt
     1441 gtcaagctct gcaccggcca tgagtatgca gctaggatca ttaacaccaa gaagctgtca
     1501 gctagagatc accagaagct ggagagggag gctcggatct gccgcctgct gaagcattcc
     1561 aacattgtac gcctccatga cagcatctct gaagagggct tccactacct ggtcttcgac
     1621 ctggtcactg gtggggagct ctttgaagac attgtggcga gagagtacta cagtgaggct
     1681 gacgccagtc actgtatcca gcagatcctg gaggctgttc tccattgtca ccaaatgggg
     1741 gtcgtccaca gagacctcaa gcctgaaaac ctgctcctgg ccagcaaatg caaaggggcc
     1801 gcagtgaaac tggcagactt cggcctggcc atcgaggttc agggagacca gcaggcatgg
     1861 tttggatttg cgggaacacc aggctacctg tctcccgaag ttcttcggaa ggaggcctat
     1921 ggcaaaccag tggatatctg ggcatgtggg gtgatcctgt atatcctgct ggtgggatac
     1981 ccacctttct gggatgagga ccagcacaag ctgtaccagc agatcaaggc tggggcctat
     2041 gacttcccat cccccgagtg ggacaccgtt acccctgaag ccaaaaacct catcaaccag
     2101 atgttgacca tcaaccccgc caagcgcatc acggcccacg aggccctgaa gcacccatgg
     2161 gtctgccaac gatccacggt ggcctccatg atgcacagac aggagactgt ggaatgtctg
     2221 aagaagttca atgcaaggag gaagctcaag ggagccatcc tcaccactat gctggccaca
     2281 cggaatttct cagtgggcag acagaccacc gctccggcca caatgtccac cgcggcctcc
     2341 ggcaccacca tggggctggt ggaacaagcc aagagtttac tcaacaagaa agcagacgga
     2401 gtcaagcccc agacaaacag caccaaaaac agctcggcca tcaccagccc caaaggatcc
     2461 ctccctcctg ccgccctgga gcctcaaacc accgttatcc ataacccagt ggacggcatt
     2521 aaggaatctt ccgacagcac caacacaacc atagaggacg aagatgccaa agcccggaag
     2581 caggaaatca tcaagaccac agagcagctc atcgaggccg tcaacaacgg cgactttgag
     2641 gcctatgcga aaatctgtga cccaggcctg acctcatttg agcctgaagc tctgggcaac
     2701 ctggtcgaag ggatggattt ccacagattc tactttgaga acctgctggc caagaacagc
     2761 aagccgatcc acaccactat cctgaacccg cacgtgcacg tcatcggcga ggatgcagcc
     2821 tgcatcgctt acatccgcct cacacagtac atcgacggcc agggcagacc ccgcaccagc
     2881 cagtccgaag agacccgtgt gtggcaccgc cgcgacggca agtggcagaa tgtccatttc
     2941 cactgctcgg gcgctccagt ggccccactg cagtaatcta gataactgat cataatcagc
     3001 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     3061 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     3121 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     3181 agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat
     3241 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     3301 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     3361 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     3421 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     3481 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     3541 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     3601 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     3661 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3721 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3781 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     3841 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3901 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3961 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     4021 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4081 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     4141 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     4201 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     4261 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     4321 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     4381 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     4441 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     4501 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     4561 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     4621 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4681 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     4741 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4801 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4861 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     4921 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     4981 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     5041 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     5101 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     5161 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     5221 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     5281 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     5341 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     5401 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     5461 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5521 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5581 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5641 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5701 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     5761 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5821 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5881 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5941 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6001 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6061 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6121 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6181 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6241 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     6301 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     6361 gttccg
//
