LOCUS       sequence-254295-        6317 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To visualize Doc2beta 6X mutant in PC12 cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6317)
  AUTHORS   Courtney NA, Briguglio JS, Bradberry MM, Greer C, Chapman ER
  TITLE     Excitatory and Inhibitory Neurons Utilize Different Ca(2+) Sensors 
            and Sources to Regulate Spontaneous Release.
  JOURNAL   Neuron. 2018 Jun 6;98(5):977-991.e5. doi: 
            10.1016/j.neuron.2018.04.022. Epub 2018 May 10.
  PUBMED    29754754
REFERENCE   2  (bases 1 to 6317)
  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"; doi: 
            "10.1016/j.neuron.2018.04.022"; journalName: "Neuron"; date: 
            "2018-06-6- 6"; volume: "98"; issue: "5"; pages: "977-991.e5"
COMMENT     Sequence Label: pEF Doc2beta 6X mutant GFP
FEATURES             Location/Qualifiers
     source          1..6317
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..1187
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          236..1178
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1135..1155
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             join(2505..2507,2508..2510,2511..3224)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2505..2507,2508..2510,2511..3221)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2550..2571)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2811..2830)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3158..3179
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3308..3327)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3373..3428
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3427..3446)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3789..3805)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3789..3805)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3802..3824)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3813..3829
                     /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(join(3837..3843,3844..3861,3862..3867))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3882..3903
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3962..4157
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4008..4143
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4163..4297
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4200..4219)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4254..4273
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4365..4382)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4536..5124)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4616..4635)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5295..6086,6087..6155))
                     /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     5918..5937
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6156..6260)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtgaggctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt
       61 ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg taaactggga
      121 aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag
      181 tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggtaag
      241 tgccgtgtgt ggttcccgcg ggcctggcct ctttacgggt tatggccctt gcgtgccttg
      301 aattacttcc acgcccctgg ctgcagtacg tgattcttga tcccgagctt cgggttggaa
      361 gtgggtggga gagttcgagg ccttgcgctt aaggagcccc ttcgcctcgt gcttgagttg
      421 aggcctggcc tgggcgctgg ggccgccgcg tgcgaatctg gtggcacctt cgcgcctgtc
      481 tcgctgcttt cgataagtct ctagccattt aaaatttttg atgacctgct gcgacgcttt
      541 ttttctggca agatagtctt gtaaatgcgg gccaagatct gcacactggt atttcggttt
      601 ttggggccgc gggcggcgac ggggcccgtg cgtcccagcg cacatgttcg gcgaggcggg
      661 gcctgcgagc gcggccaccg agaatcggac gggggtagtc tcaagctggc cggcctgctc
      721 tggtgcctgg cctcgcgccg ccgtgtatcg ccccgccctg ggcggcaagg ctggcccggt
      781 cggcaccagt tgcgtgagcg gaaagatggc cgcttcccgg ccctgctgca gggagctcaa
      841 aatggaggac gcggcgctcg ggagagcggg cgggtgagtc acccacacaa aggaaaaggg
      901 cctttccgtc ctcagccgtc gcttcatgtg actccacgga gtaccgggcg ccgtccaggc
      961 acctcgatta gttctcgagc ttttggagta cgtcgtcttt aggttggggg gaggggtttt
     1021 atgcgatgga gtttccccac actgagtggg tggagactga agttaggcca gcttggcact
     1081 tgatgtaatt ctccttggaa tttgcccttt ttgagtttgg atcttggttc attctcaagc
     1141 ctcagacagt ggttcaaagt ttttttcttc catttcaggt gtcgtgagga attctgcagt
     1201 cgacggtacc gcgggcccgg gccaccatga ccctcaggag gcggggggag aaggcaacca
     1261 tttccattca agagcatatg gctattgatg tctgccctgg gccaatacgg cctattaagc
     1321 aaataagcga ttacttccca cgatttccac gaggactgcc acctaccgca gccccaaggg
     1381 cttccgcccc cccagacgcc cccgctcggt cccctgcagc cacagcaggc ccacggtcac
     1441 cttctgatgg ggcccgagat gacgatgaag atgtcgacca gcttttcgga gcttacggag
     1501 cctcccccgg tccctcacca ggtccttctc ctgtgcgccc acctgccaag ccccccgaag
     1561 acgaacctga tgctgatggt tacgagagcg acgattgtac cgcattgggt actctggact
     1621 tcagcctcct gtacgatcaa gagaataatg cacttcactg cactatttcc aaggctaaag
     1681 gcctcaagcc tatggatcat aacggcttgg ccgctcccta cgtcaagctc catttgcttc
     1741 ccggggctag taaggctaac aagctccgga caaaaacatt gagaaataca ctgaatccat
     1801 cctggaacga gacactcaca tattatggta tcaccgatga agacatgata cgcaaaactc
     1861 tgagaatctc tgtctgtgcc gaagctaaat ttaggcacaa tgaattcatt ggcgagactc
     1921 gagtacccct caaaaagttg aaacccaatc atactaagac ctttagcatc tgtcttgaaa
     1981 agcaactccc tgtggataag gctgaagata aatcactcga ggaaaggggg cgcatcctca
     2041 tctccctcaa atattcttca caaaaacaag gccttctggt tggaatcgtc aggtgcgcac
     2101 atctggcagc tatggacgct aacggatatt ctgctcccta tgttaagaca tatctcaaac
     2161 cagacgttga caaaaaaagt aaacataaaa ctgctgtcaa gaagaaaaca ttgaatccag
     2221 aatttaatga ggagttttgc tacgaaatca aacatggtga tcttgctaag aagactctcg
     2281 aggtaaccgt ctgggcctat gccatcggga aaagtaacga ttttataggt ggagtcgttc
     2341 tcggaataaa tgccaagggg gaaaggctga agcattggtt cgactgcttg aagaacaagg
     2401 ataagagaat cgaacgatgg cacactctta caaacgagat cccaggtgcc gttctcagtg
     2461 atggatccgg aagttcgggg tccagcggtt cttcaggcag ttccatggtg agcaagggcg
     2521 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     2581 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     2641 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     2701 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     2761 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     2821 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     2881 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     2941 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     3001 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     3061 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     3121 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     3181 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaggccgc actcctcagg
     3241 tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa tgccctggct cacaaatacc
     3301 actgagatct ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc
     3361 tgacttctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt
     3421 gtctctcact cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt
     3481 tggtttagag tttggcaaca tatgcccata tgctggctgc catgaacaaa ggttggctat
     3541 aaagaggtca tcagtatatg aaacagcccc ctgctgtcca ttccttattc catagaaaag
     3601 ccttgacttg aggttagatt ttttttatat tttgttttgt gttatttttt tctttaacat
     3661 ccctaaaatt ttccttacat gttttactag ccagattttt cctcctctcc tgactactcc
     3721 cagtcatagc tgtccctctt ctcttatgga gatccctcga cctgcagccc aagcttggcg
     3781 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     3841 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     3901 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc
     3961 cgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4021 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4081 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4141 gcctaggctt ttgcaaaaag ctaacttgtt tattgcagct tataatggtt acaaataaag
     4201 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     4261 gtccaaactc atcaatgtat cttatcatgt ctggatccgc tgcattaatg aatcggccaa
     4321 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4381 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4441 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4501 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4561 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4621 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4681 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4741 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4801 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4861 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4921 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4981 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5041 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5101 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5161 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5221 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5281 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5341 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5401 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5461 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5521 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5581 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5641 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5701 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5761 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5821 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5881 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5941 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6001 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6061 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6121 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6181 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6241 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgggtc gacattgatt
     6301 attgactaga tcatcgc
//
