LOCUS       sequence-177820-        5902 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FRET probe; extracellular synapse targeted mCherry, Cre-dependently 
            expressed.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5902)
  AUTHORS   Octeau JC, Chai H, Jiang R, Bonanno SL, Martin KC, Khakh BS
  TITLE     An Optical Neuron-Astrocyte Proximity Assay at Synaptic Distance 
            Scales.
  JOURNAL   Neuron. 2018 Apr 4;98(1):49-66.e9. doi: 
            10.1016/j.neuron.2018.03.003.
  PUBMED    29621490
REFERENCE   2  (bases 1 to 5902)
  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.2018.03"; journalName: "Neuron"; date: "2018-04-4-
            4"; volume: "98"; issue: "1"; pages: "49-66.e9"
COMMENT     Sequence Label: pZac2.1 hSynapsin1 FLEX NAPA-N SV40
FEATURES             Location/Qualifiers
     source          1..5902
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   259..388
                     /label=AAV2 ITR
     promoter        465..912
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    934..967
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    1030..1063
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1928..2635)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2203..2222)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2472..2490
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     protein_bind    complement(2735..2768)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(2839..2872)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    2927..3048
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2938..2957)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2992..3011
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3210..3339
                     /label=AAV2 ITR
     primer_bind     complement(3358..3375)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3358..3374)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3367..3389)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3516..3971
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3603..3622)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3813..3834
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3997..4101
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4102..4170,4171..4962)
                     /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     complement(4320..4339)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5133..5721
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5622..5641
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccagattta
      241 attaaggcct taattaggct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc
      301 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
      361 gtggccaact ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta
      421 tctacgtagc catgctctag gaagatctga gggccctgcg tatgagtgca agtgggtttt
      481 aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga cccactggac
      541 aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg gaggggaaac
      601 aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt gccttcgccc
      661 ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac gtcactcgcc
      721 ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc cgccggccca
      781 gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca tctgcgctgc
      841 ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt cgtgtcgtgc
      901 ctgagagcgc agtcgagaga acttaagctg cagataactt cgtataaagt atcctatacg
      961 aagttatatc aaaataggaa gaccaatgct tcaccatcga cccgaattgc caagcatcac
     1021 catcgaccca taacttcgta taatgtatgc tatacgaagt tatactagtt tatacgtagt
     1081 actctttatc cttgtttttc ttgtttttgt ttgctgattt ggcgctggag ggctgtttct
     1141 ctttaaccac ggcgccatta ctctgagcgc tgttactgat atagttccta gattcgtcaa
     1201 cgtggtaaga tccctcatcc ctgttgcgat acttgtagcg tacgcagaag aagatgccca
     1261 ggccgataaa cagaagcagg ccggcgactc ccccgagtac gatcagggcc atcggctgta
     1321 ctggtgttga ccaagtagga agcaccttga tgttgctctc cagcagcacc tgtccagagt
     1381 cgctaagaag acactgccac atccctgcct ccgggttaag gacccacaca gccttctctc
     1441 ttttactgac tttagcctcc ttattctcca gcttcagaga cagcatcagt ttaggtgagg
     1501 tgggacccca cacttcgcag gtcaggttct tctgcagctg tgttgccctc atcaccacga
     1561 ggttgacctc ctgatgcaac tttccggtct tagcctccaa ggcgagggtg aggttcccgg
     1621 agccagcata ctggggcagg gcttggggga gtgtcaagtg caaaggcaac ttcttgccca
     1681 tctgcagctt agggtcctgt gtgactctct tcacgctcac ttctttgttt ttcagatcaa
     1741 aagtgatcca actcttgctg gaggaggctc tttcggcctg ccaccacaac tctcctgatc
     1801 ctgtgagctt ctcgactgtg aaggccaatg ggaaactgaa ctccacctgt tccccctcct
     1861 tcttataaac aatagagcta gctttctgaa agtcgacgct accaccgccg ccagatccgc
     1921 caccgccctt gtacagctcg tccatgccgc cggtggagtg gcggccctcg gcgcgttcgt
     1981 actgttccac gatggtgtag tcctcgttgt gggaggtgat gtccaacttg atgttgacgt
     2041 tgtaggcgcc gggcagctgc acgggcttct tggccttgta ggtggtcttg acctcagcgt
     2101 cgtagtggcc gccgtccttc agcttcagcc tctgcttgat ctcgcccttc agggcgccgt
     2161 cctcggggta catccgctcg gaggaggcct cccagcccat ggtcttcttc tgcattacgg
     2221 ggccgtcgga ggggaagttg gtgccgcgca gcttcacctt gtagatgaac tcgccgtcct
     2281 gcagggagga gtcctgggtc acggtcacca cgccgccgtc ctcgaagttc atcacgcgct
     2341 cccacttgaa gccctcgggg aaggacagct tcaagtagtc ggggatgtcg gcggggtgct
     2401 tcacgtaggc cttggagccg tacatgaact gaggggacag gatgtcccag gcgaagggca
     2461 gggggccacc cttggtcacc ttcagcttgg cggtctgggt gccctcgtag gggcggccct
     2521 cgccctcgcc ctcgatctcg aactcgtggc cgttcacgga gccctccatg tgcaccttga
     2581 agcgcatgaa ctccttgatg atggccatgt tatcctcctc gcccttgctc accatggtga
     2641 cttctcctgt cttccaatct gaagcgggaa gagcgaaggg gagaagtgca gcaagcaaca
     2701 aaagactagt agaaggtggc atggtggtac tagtataact tcgtatagga tactttatac
     2761 gaagttatca ttgggattct tcctattttg atccaagcat caccatcgac cctctagtcc
     2821 agatctcacc atcgacccat aacttcgtat agcatacatt atacgaagtt atgtccctcg
     2881 aagaggttct agagtcgacc cgggcggccg cttcgagcag acatgataag atacattgat
     2941 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     3001 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     3061 tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggtttttta aagcaagtaa
     3121 aacctctaca aatgtggtaa aatcgataag gatcttccta gagcatggct acgtagataa
     3181 gtagcatggc gggttaatca ttaactacaa ggaaccccta gtgatggagt tggccactcc
     3241 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     3301 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc cttaattaac ctaattcact
     3361 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     3421 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     3481 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     3541 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     3601 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3661 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3721 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3781 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3841 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     3901 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     3961 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4021 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4081 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     4141 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     4201 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     4261 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     4321 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     4381 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     4441 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     4501 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     4561 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     4621 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     4681 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     4741 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     4801 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     4861 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     4921 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     4981 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5041 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5101 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5161 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5221 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     5281 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     5341 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     5401 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     5461 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     5521 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     5581 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     5641 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     5701 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     5761 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     5821 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     5881 tcagtgagcg aggaagcgga ag
//
