LOCUS       sequence-370179-        5958 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Synaptic buton marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5958)
  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 5958)
  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-mRuby
FEATURES             Location/Qualifiers
     source          1..5958
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        53..1235
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          284..1226
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1183..1203
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1246..1255
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2997..3016)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3062..3117
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3116..3135)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3478..3494)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3478..3494)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3491..3513)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3502..3518
                     /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(3526..3532,3533..3550,3551..3556))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3571..3592
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3651..3846
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3697..3832
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3852..3986
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3889..3908)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3943..3962
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4054..4071)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4225..4813)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4305..4324)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4984..5775,5776..5844))
                     /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     5607..5626
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5845..5949)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccgaaaagtg ccacctgggt cgacattgat tattgactag atcatcgcgt gaggctccgg
       61 tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt
      121 cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt
      181 gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc
      241 cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg
      301 ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa ttacttccac
      361 gcccctggct gcagtacgtg attcttgatc ccgagcttcg ggttggaagt gggtgggaga
      421 gttcgaggcc ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg
      481 ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg
      541 ataagtctct agccatttaa aatttttgat gacctgctgc gacgcttttt ttctggcaag
      601 atagtcttgt aaatgcgggc caagatctgc acactggtat ttcggttttt ggggccgcgg
      661 gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc gaggcggggc ctgcgagcgc
      721 ggccaccgag aatcggacgg gggtagtctc aagctggccg gcctgctctg gtgcctggcc
      781 tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg gcaccagttg
      841 cgtgagcgga aagatggccg cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc
      901 ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct
      961 cagccgtcgc ttcatgtgac tccacggagt accgggcgcc gtccaggcac ctcgattagt
     1021 tctcgagctt ttggagtacg tcgtctttag gttgggggga ggggttttat gcgatggagt
     1081 ttccccacac tgagtgggtg gagactgaag ttaggccagc ttggcacttg atgtaattct
     1141 ccttggaatt tgcccttttt gagtttggat cttggttcat tctcaagcct cagacagtgg
     1201 ttcaaagttt ttttcttcca tttcaggtgt cgtgaggaat tgcccgccac catggacgtg
     1261 gtgaatcagc tggtggctgg gggtcagttc cgggtggtca aggagcccct tggcttcgtg
     1321 aaggtgctgc agtgggtctt tgccatcttc gcctttgcta cgtgtggcag ctacaccggg
     1381 gagcttcggc tgagcgtgga gtgtgccaac aagacggaga gtgccctcaa catcgaagtt
     1441 gaattcgagt accccttcag gctgcaccaa gtgtactttg atgcaccctc ctgcgtcaaa
     1501 gggggcacta ccaagatctt cctggttggg gactactcct cgtcggctga attctttgtc
     1561 accgtggctg tgtttgcctt cctctactcc atgggggccc tggccaccta catcttcctg
     1621 cagaacaagt accgagagaa caacaaaggg cctatgatgg actttctggc tacagccgtg
     1681 ttcgctttca tgtggctagt tagttcatca gcctgggcca aaggcctgtc cgatgtgaag
     1741 atggccacgg acccagagaa cattatcaag gagatgccca tgtgccgcca gacagggaac
     1801 acatgcaagg aactgaggga ccctgtgact tcaggactca acacctcagt ggtgtttggc
     1861 ttcctgaacc tggtgctctg ggttggcaac ttatggttcg tgttcaagga gacaggctgg
     1921 gcagccccat tcatgcgcgc acctccaggc gccccggaaa agcaaccagc acctggcgat
     1981 gcctacggcg atgcgggcta cgggcagggc cccggaggct atgggcccca ggactcctac
     2041 gggcctcagg gtggttatca acccgattac gggcagccag ccagcggtgg cggtggctac
     2101 gggcctcagg gcgactatgg gcagcaaggc tatggccaac agggtgcgcc cacctccttc
     2161 tccaatcaga tgaaaggatc cggaagttcg gggtccagcg gttcttcagg cagttccatg
     2221 gtgtctaagg gcgaagagct gatcaaggaa aatatgcgta tgaaggtggt catggaaggt
     2281 tcggtcaacg gccaccaatt caaatgcaca ggtgaaggag aaggcagacc gtacgaggga
     2341 gtgcaaacca tgaggatcaa agtcatcgag ggaggacccc tgccatttgc ctttgacatt
     2401 cttgccacgt cgttcatgta tggcagccgt acctttatca agtacccggc cgacatccct
     2461 gatttcttta aacagtcctt tcctgagggt tttacttggg aaagagttac gagatacgaa
     2521 gatggtggag tcgtcaccgt cacgcaggac accagccttg aggatggcga gctcgtctac
     2581 aacgtcaagg tcagaggggt aaactttccc tccaatggtc ccgtgatgca gaagaagacc
     2641 aagggttggg agcctaatac agagatgatg tatccagcag atggtggtct gagaggatac
     2701 actgacatcg cactgaaagt tgatggtggt ggccatctgc actgcaactt cgtgacaact
     2761 tacaggtcaa aaaagaccgt cgggaacatc aagatgcccg gtgtccatgc cgttgatcac
     2821 cgcctggaaa ggatcgagga gagtgacaat gaaacctacg tagtgcaaag agaagtggca
     2881 gttgccaaat acagcaacct tggtggtggc tagggccgca ctcctcaggt gcaggctgcc
     2941 tatcagaagg tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt
     3001 tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc
     3061 taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc
     3121 ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt
     3181 ttggcaacat atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat
     3241 cagtatatga aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga
     3301 ggttagattt tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt
     3361 tccttacatg ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct
     3421 gtccctcttc tcttatggag atccctcgac ctgcagccca agcttggcgt aatcatggtc
     3481 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     3541 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     3601 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat
     3661 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3721 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3781 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3841 tgcaaaaagc taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3901 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3961 tcaatgtatc ttatcatgtc tggatccgct gcattaatga atcggccaac gcgcggggag
     4021 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4081 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4141 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4201 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4261 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4321 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4381 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4441 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4501 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4561 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4621 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4681 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4741 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4801 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4861 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4921 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4981 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5041 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5101 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     5161 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     5221 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     5281 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5341 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5401 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5461 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5521 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5581 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5641 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5701 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5761 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5821 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     5881 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     5941 ggttccgcgc acatttcc
//
