LOCUS       sequence-254294-        6311 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To visualize Doc2beta WT in PC12 cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6311)
  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 6311)
  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 WT GFP
FEATURES             Location/Qualifiers
     source          1..6311
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          231..1173
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1130..1150
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             join(2494..2496,2497..2499,2500..3213)
                     /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(2494..2496,2497..2499,2500..3210)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2539..2560)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2800..2819)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3147..3168
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3297..3316)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3362..3417
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3416..3435)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3778..3794)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3778..3794)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3791..3813)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3802..3818
                     /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(3826..3832,3833..3850,3851..3856))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3871..3892
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3951..4146
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3997..4132
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4152..4286
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4189..4208)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4243..4262
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4354..4371)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4525..5113)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4605..4624)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5284..6075,6076..6144))
                     /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     5907..5926
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6145..6249)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(6311,1..1182)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
ORIGIN
        1 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
       61 gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
      121 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
      181 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg
      241 tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta
      301 cttccacgcc cctggctgca gtacgtgatt cttgatcccg agcttcgggt tggaagtggg
      361 tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc
      421 tggcctgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct
      481 gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac gctttttttc
      541 tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc ggtttttggg
      601 gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg
      661 cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc tgctctggtg
      721 cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc ccggtcggca
      781 ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag ctcaaaatgg
      841 aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa aagggccttt
      901 ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc caggcacctc
      961 gattagttct cgagcttttg gagtacgtcg tctttaggtt ggggggaggg gttttatgcg
     1021 atggagtttc cccacactga gtgggtggag actgaagtta ggccagcttg gcacttgatg
     1081 taattctcct tggaatttgc cctttttgag tttggatctt ggttcattct caagcctcag
     1141 acagtggttc aaagtttttt tcttccattt caggtgtcgt gaggaattct gcagtcgacg
     1201 gtaccgcggg cccggatgac cctccgaaga cgtggagaga aggctacaat cagcatccaa
     1261 gagcatatgg caatcgacgt gtgtcctgga cctatccgac ctatcaagca gatctccgat
     1321 tacttcccac gattccctcg tggactccca ccaacagctg caccacgtgc ttcagcacct
     1381 ccagacgctc ctgcacgatc gccagcagct accgcaggtc ctcgtagccc ctccgacgga
     1441 gcacgcgacg acgacgaaga tgtggaccag ctcttcggag cctacggtgc cagcccaggc
     1501 cccagccccg gacccagccc cgtgaggccg ccggccaagc cccccgagga cgaaccggac
     1561 gccgacggct acgagtcaga cgactgcacc gccctgggta cactggactt cagtctgctc
     1621 tatgaccagg agaacaacgc actgcactgc accatcagca aagccaaggg cctgaagccg
     1681 atggatcaca atggactggc tgatccctac gtcaaactac acttgctgcc tggagccagc
     1741 aaggcaaata agctcagaac aaaaacccta cggaacactc tgaacccctc ctggaacgag
     1801 accctcacct attacgggat cacggatgag gacatgatcc gaaagaccct gaggatctcc
     1861 gtgtgtgacg aggacaaatt ccgccacaac gagttcatcg gagagactcg agtgcccctg
     1921 aagaagctga aacccaacca caccaagaca ttcagcatct gcctggagaa gcagctgccg
     1981 gtggacaagg cagaggacaa gtccctggag gagcgtggcc gcatcctcat ctctctcaag
     2041 tacagctcac agaagcaggg cctgctggtg ggcatcgttc gctgcgcaca cctggcggcc
     2101 atggatgcta acggctactc agacccctac gtgaaaacat atctgaaacc agatgtagac
     2161 aagaaatcca agcataagac cgcagtcaag aagaaaacac taaaccctga attcaatgag
     2221 gagttctgtt acgagatcaa gcatggagac ctagccaaaa agaccctgga ggtcactgtc
     2281 tgggactatg atattggaaa atccaatgat ttcatcggtg gggtggttct gggcatcaat
     2341 gccaagggtg agcgcctgaa gcactggttt gactgtctga agaacaagga caagaggatt
     2401 gagcgttggc acacgctcac caatgagatc ccaggggctg tactcagcga cggatccgga
     2461 agttcggggt ccagcggttc ttcaggcagt tccatggtga gcaagggcga ggagctgttc
     2521 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     2581 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     2641 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     2701 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     2761 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     2821 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     2881 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     2941 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     3001 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     3061 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     3121 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     3181 atcactctcg gcatggacga gctgtacaag taaggccgca ctcctcaggt gcaggctgcc
     3241 tatcagaagg tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt
     3301 tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc
     3361 taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc
     3421 ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt
     3481 ttggcaacat atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat
     3541 cagtatatga aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga
     3601 ggttagattt tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt
     3661 tccttacatg ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct
     3721 gtccctcttc tcttatggag atccctcgac ctgcagccca agcttggcgt aatcatggtc
     3781 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     3841 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     3901 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat
     3961 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     4021 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     4081 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     4141 tgcaaaaagc taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     4201 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     4261 tcaatgtatc ttatcatgtc tggatccgct gcattaatga atcggccaac gcgcggggag
     4321 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4381 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4441 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4501 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4561 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4621 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4681 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4741 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4801 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4861 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4921 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4981 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5041 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5101 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5161 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5221 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5281 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5341 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5401 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     5461 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     5521 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     5581 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5641 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5701 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5761 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5821 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5881 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5941 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6001 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6061 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6121 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6181 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6241 ggttccgcgc acatttcccc gaaaagtgcc acctgggtcg acattgatta ttgactagat
     6301 catcgcgtga g
//
