LOCUS       sequence-254208-        6284 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To visualize Doc2alpha WT in PC12 cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6284)
  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 6284)
  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 Doc2alpha WT GFP
FEATURES             Location/Qualifiers
     source          1..6284
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        62..1244
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          293..1235
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1192..1212
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             join(2529..2531,2532..2534,2535..3248)
                     /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(2529..2531,2532..2534,2535..3245)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2574..2595)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2835..2854)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3182..3203
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3332..3351)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3397..3452
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3451..3470)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3813..3829)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3813..3829)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3826..3848)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3837..3853
                     /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(3861..3867,3868..3885,3886..3891))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3906..3927
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3986..4181
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4032..4167
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4187..4321
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4224..4243)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4278..4297
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4389..4406)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4560..5148)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4640..4659)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5319..6110,6111..6179))
                     /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     5942..5961
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6180..6284)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacatttccc cgaaaagtgc cacctgggtc gacattgatt attgactaga tcatcgcgtg
       61 aggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
      121 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
      181 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
      241 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
      301 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
      361 tacttccacg cccctggctg cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg
      421 ggtgggagag ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct tgagttgagg
      481 cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg
      541 ctgctttcga taagtctcta gccatttaaa atttttgatg acctgctgcg acgctttttt
      601 tctggcaaga tagtcttgta aatgcgggcc aagatctgca cactggtatt tcggtttttg
      661 gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc
      721 tgcgagcgcg gccaccgaga atcggacggg ggtagtctca agctggccgg cctgctctgg
      781 tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg
      841 caccagttgc gtgagcggaa agatggccgc ttcccggccc tgctgcaggg agctcaaaat
      901 ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct
      961 ttccgtcctc agccgtcgct tcatgtgact ccacggagta ccgggcgccg tccaggcacc
     1021 tcgattagtt ctcgagcttt tggagtacgt cgtctttagg ttggggggag gggttttatg
     1081 cgatggagtt tccccacact gagtgggtgg agactgaagt taggccagct tggcacttga
     1141 tgtaattctc cttggaattt gccctttttg agtttggatc ttggttcatt ctcaagcctc
     1201 agacagtggt tcaaagtttt tttcttccat ttcaggtgtc gtgaggaatt ctgcagtcga
     1261 cggtaccgcg ggcccggatg agaggacgaa gaggtgatcg aatgaccatc aacatccagg
     1321 agcacatggc aatcaacgtg tgccctggac ccatcaggcc catccgccag atctccgact
     1381 acttccctcg cagaggacca ggaccagagg gtggcggcgg aggaaggggc ttcggggaag
     1441 ctccagctca tctggcccct ctggctctgg ctccacctgc agctctcctt ggagccacta
     1501 cacccgacga tggagctgag gtagacagct acgactcgga tgataccact gccctgggca
     1561 cgctggaatt tgaccttctc tatgatcagg cttcctgcat gctgcactgt agaatcctca
     1621 gggccaaggg cctcaagccc atggatttca acggcctggc tgacccctat gtaaagctgc
     1681 acctcctgcc aggagcctgc aaggccaata agctaaaaac caagacacag aggaacacac
     1741 tgaacccagt gtggaacgag gagctgacgt acagtggaat cacagatgac gacatcaccc
     1801 acaaggtgct caggatctcg gtctgtgatg aggacaagct gagtcacaat gaatttattg
     1861 gggagatccg agtgcccctc cgccgcctca agccttcaca gaagaagcat tttaacatct
     1921 gccttgagcg tcaggtcccg cttccttcgc cctcttcaat gtctgcggcg ctgaggggca
     1981 tatcctgtta cctgaaggag ctagagcagg cagagcaggg acctgggctg ttggaagagc
     2041 gcgggcgcat cctgctgagc ctcagctaca gctctcggcg gcatgggctg ctggtgggca
     2101 ttgttcgctg tgctcacctg gctgcaatgg atgttaacgg ctactctgac ccttatgtga
     2161 agacgtactt gagaccagac gtggataaga aatccaagca taagacgtgt gtaaagaaga
     2221 agacgctgaa cccggaattt aatgaggaat tcttttatga gatggaactc tccactctgg
     2281 ctaccaagac cctggaagtc acagtctggg actacgacat tggcaaatcc aatgacttca
     2341 taggtggtgt gtctctgggg ccaggagccc ggggagaggc ccagaaacac tggagagact
     2401 gtctacatca gccggacaca gcagtggagc gctggcatac cctgaccagc gagctgcccc
     2461 cagcagcagg ggcgttgccg ttggccggat ccggaagttc ggggtccagc ggttcttcag
     2521 gcagttccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     2581 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     2641 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     2701 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     2761 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     2821 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     2881 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     2941 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     3001 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     3061 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     3121 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     3181 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     3241 acaagtaagg ccgcactcct caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg
     3301 ccaatgccct ggctcacaaa taccactgag atctttttcc ctctgccaaa aattatgggg
     3361 acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt attttcattg
     3421 caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc
     3481 atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg
     3541 ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg
     3601 tccattcctt attccataga aaagccttga cttgaggtta gatttttttt atattttgtt
     3661 ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat
     3721 ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta tggagatccc
     3781 tcgacctgca gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     3841 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     3901 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     3961 ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc agcaaccata gtcccgcccc
     4021 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     4081 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     4141 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctaact tgtttattgc
     4201 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4261 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat
     4321 ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4381 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4441 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4501 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4561 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4621 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4681 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4741 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4801 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4861 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     4921 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     4981 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5041 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5101 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5161 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5221 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5281 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5341 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5401 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     5461 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5521 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     5581 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     5641 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5701 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     5761 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     5821 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     5881 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     5941 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6001 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6061 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6121 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6181 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6241 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcg
//
