LOCUS       sequence-379038-        6524 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Presynaptic fluorescent  Ca2+ indicator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6524)
  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 6524)
  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 Synaptophysin-GCamp6s
FEATURES             Location/Qualifiers
     source          1..6524
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        79..1261
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          310..1252
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1209..1229
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1272..1281
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2247..2256
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2268..2324
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     2537..2558
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2664..2685)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2925..2944)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(3589..3608)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3654..3709
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3708..3727)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4070..4086)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4070..4086)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4083..4105)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4094..4110
                     /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(4118..4124,4125..4142,4143..4148))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4163..4184
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4243..4438
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4289..4424
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4444..4578
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4481..4500)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4535..4554
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4646..4663)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4817..5405)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4897..4916)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5576..6367,6368..6436))
                     /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     6199..6218
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(6437..6524,1..17))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgggtcgac attgattatt
       61 gactagatca tcgcgtgagg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca
      121 gtccccgaga agttgggggg aggggtcggc aattgaaccg gtgcctagag aaggtggcgc
      181 ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga
      241 gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc
      301 agaacacagg taagtgccgt gtgtggttcc cgcgggcctg gcctctttac gggttatggc
      361 ccttgcgtgc cttgaattac ttccacgccc ctggctgcag tacgtgattc ttgatcccga
      421 gcttcgggtt ggaagtgggt gggagagttc gaggccttgc gcttaaggag ccccttcgcc
      481 tcgtgcttga gttgaggcct ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca
      541 ccttcgcgcc tgtctcgctg ctttcgataa gtctctagcc atttaaaatt tttgatgacc
      601 tgctgcgacg ctttttttct ggcaagatag tcttgtaaat gcgggccaag atctgcacac
      661 tggtatttcg gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg
      721 ttcggcgagg cggggcctgc gagcgcggcc accgagaatc ggacgggggt agtctcaagc
      781 tggccggcct gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc
      841 aaggctggcc cggtcggcac cagttgcgtg agcggaaaga tggccgcttc ccggccctgc
      901 tgcagggagc tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg agtcacccac
      961 acaaaggaaa agggcctttc cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg
     1021 ggcgccgtcc aggcacctcg attagttctc gagcttttgg agtacgtcgt ctttaggttg
     1081 gggggagggg ttttatgcga tggagtttcc ccacactgag tgggtggaga ctgaagttag
     1141 gccagcttgg cacttgatgt aattctcctt ggaatttgcc ctttttgagt ttggatcttg
     1201 gttcattctc aagcctcaga cagtggttca aagttttttt cttccatttc aggtgtcgtg
     1261 aggaattgcc cgccaccatg gacgtggtga atcagctggt ggctgggggt cagttccggg
     1321 tggtcaagga gccccttggc ttcgtgaagg tgctgcagtg ggtctttgcc atcttcgcct
     1381 ttgctacgtg tggcagctac accggggagc ttcggctgag cgtggagtgt gccaacaaga
     1441 cggagagtgc cctcaacatc gaagttgaat tcgagtaccc cttcaggctg caccaagtgt
     1501 actttgatgc accctcctgc gtcaaagggg gcactaccaa gatcttcctg gttggggact
     1561 actcctcgtc ggctgaattc tttgtcaccg tggctgtgtt tgccttcctc tactccatgg
     1621 gggccctggc cacctacatc ttcctgcaga acaagtaccg agagaacaac aaagggccta
     1681 tgatggactt tctggctaca gccgtgttcg ctttcatgtg gctagttagt tcatcagcct
     1741 gggccaaagg cctgtccgat gtgaagatgg ccacggaccc agagaacatt atcaaggaga
     1801 tgcccatgtg ccgccagaca gggaacacat gcaaggaact gagggaccct gtgacttcag
     1861 gactcaacac ctcagtggtg tttggcttcc tgaacctggt gctctgggtt ggcaacttat
     1921 ggttcgtgtt caaggagaca ggctgggcag ccccattcat gcgcgcacct ccaggcgccc
     1981 cggaaaagca accagcacct ggcgatgcct acggcgatgc gggctacggg cagggccccg
     2041 gaggctatgg gccccaggac tcctacgggc ctcagggtgg ttatcaaccc gattacgggc
     2101 agccagccag cggtggcggt ggctacgggc ctcagggcga ctatgggcag caaggctatg
     2161 gccaacaggg tgcgcccacc tccttctcca atcagatgaa aggatccgga agttcggggt
     2221 ccagcggttc ttcaggcagt tccctcgcca ccatggtcga ctcatcacgt cgtaagtgga
     2281 ataagacagg tcacgcagtc agagctatag gtcggctgag ctcactcgag aacgtctata
     2341 tcaaggccga caagcagaag aacggcatca aggcgaactt ccacatccgc cacaacatcg
     2401 aggacggcgg cgtgcagctc gcctaccact accagcagaa cacccccatc ggcgacggcc
     2461 ccgtgctgct gcccgacaac cactacctga gcgtgcagtc caaactttcg aaagacccca
     2521 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     2581 gcatggacga gctgtacaag ggcggtaccg gagggagcat ggtgagcaag ggcgaggagc
     2641 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     2701 tcagcgtgtc cggcgagggt gagggcgatg ccacctacgg caagctgacc ctgaagttca
     2761 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     2821 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     2881 ccatgcccga aggctacatc caggagcgca ccatcttctt caaggacgac ggcaactaca
     2941 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     3001 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aacctgccgg
     3061 accaactgac tgaagagcag atcgcagaat ttaaagaggc tttctcccta tttgacaagg
     3121 acggggatgg gacaataaca accaaggagc tggggacggt gatgcggtct ctggggcaga
     3181 accccacaga agcagagctg caggacatga tcaatgaagt agatgccgac ggtgacggca
     3241 caatcgactt ccctgagttc ctgacaatga tggcaagaaa aatgaaatac agggacacgg
     3301 aagaagaaat tagagaagcg ttcggtgtgt ttgataagga tggcaatggc tacatcagtg
     3361 cagcagagct tcgccacgtg atgacaaacc ttggagagaa gttaacagat gaagaggttg
     3421 atgaaatgat cagggaagca gacatcgatg gggatggtca ggtaaactac gaagagtttg
     3481 tacaaatgat gacagcgaag tgagcggccg cactcctcag gtgcaggctg cctatcagaa
     3541 ggtggtggct ggtgtggcca atgccctggc tcacaaatac cactgagatc tttttccctc
     3601 tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag
     3661 gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac
     3721 atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac
     3781 atatgcccat atgctggctg ccatgaacaa aggttggcta taaagaggtc atcagtatat
     3841 gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat
     3901 tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca
     3961 tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct
     4021 tctcttatgg agatccctcg acctgcagcc caagcttggc gtaatcatgg tcatagctgt
     4081 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4141 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4201 tgcccgcttt ccagtcggga aacctgtcgt gccagcggat ccgcatctca attagtcagc
     4261 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     4321 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     4381 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     4441 gctaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     4501 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     4561 tcttatcatg tctggatccg ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     4621 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     4681 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4741 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4801 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4861 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4921 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4981 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     5041 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5101 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5161 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5221 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     5281 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5341 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5401 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5461 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5521 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5581 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5641 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     5701 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     5761 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     5821 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     5881 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     5941 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6001 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6061 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6121 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6181 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6241 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6301 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6361 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6421 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6481 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaa
//
