LOCUS       sequence-181876-        5914 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FRET probe; extracelllular synapse targeted mScarlet.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5914)
  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 5914)
  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 NAPA-N2 SV40
FEATURES             Location/Qualifiers
     source          1..5914
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    117..138
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(153..158,159..176,177..183)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    191..207
                     /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     196..218
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     215..231
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     215..231
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   269..398
                     /label=AAV2 ITR
     promoter        475..922
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     intron          969..1101
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1084..1103
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1146..1164
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1269..1964
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(1402..1420)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1670..1689
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    1965..2030
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2949..3070
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2960..2979)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3014..3033
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3232..3361
                     /label=AAV2 ITR
     primer_bind     complement(3380..3397)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3380..3396)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3389..3411)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3538..3993
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3625..3644)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3835..3856
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4019..4123
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4124..4192,4193..4984)
                     /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(4342..4361)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5155..5743
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5644..5663
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
       61 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
      121 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
      181 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
      241 gccagattta attaaggcct taattaggct gcgcgctcgc tcgctcactg aggccgcccg
      301 ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg
      361 cagagaggga gtggccaact ccatcactag gggttccttg tagttaatga ttaacccgcc
      421 atgctactta tctacgtagc catgctctag gaagatctga gggccctgcg tatgagtgca
      481 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      541 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      601 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      661 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      721 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      781 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      841 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      901 cgtgtcgtgc ctgagagcgc agtcgagaga acttaagctg cagaagttgg tcgtgaggca
      961 ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct
     1021 tgtcgagaca gagaaaactc ttgcgtttct gataggcacc tattggtctt actgacatcc
     1081 actttgcctt tctctccaca ggtgtccact cccagttcaa ttacagctct taaggctaga
     1141 gtacttaata cgactcacta taggctagcc tcgagaccac catgccacct tctactagtc
     1201 ttttgttgct tgctgcactt ctccccttcg ctcttcccgc ttcagattgg aagacaggag
     1261 aagtcaccat ggtgagcaag ggcgaggcag tgatcaagga gttcatgcgg ttcaaggtgc
     1321 acatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     1381 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttct
     1441 cctgggacat cctgtcccct cagttcatgt acggctccag ggccttcacc aagcaccccg
     1501 ccgacatccc cgactactat aagcagtcct tccccgaggg cttcaagtgg gagcgcgtga
     1561 tgaacttcga ggacggcggc gccgtgaccg tgacccagga cacctccctg gaggacggca
     1621 ccctgatcta caaggtgaag ctccgcggca ccaacttccc tcctgacggc cccgtaatgc
     1681 agaagaagac aatgggctgg gaagcgtcca ccgagcggtt gtaccccgag gacggcgtgc
     1741 tgaagggcga cattaagatg gccctgcgcc tgaaggacgg cggccgctac ctggcggact
     1801 tcaagaccac ctacaaggcc aagaagcccg tgcagatgcc cggcgcctac aacgtcgacc
     1861 gcaagttgga catcacctcc cacaacgagg actacaccgt ggtggaacag tacgaacgct
     1921 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtccgga ctcagatctc
     1981 gagctcaagc ttcgaattct gcagtcgacg gtaccgcggg cccgggatcc accggatcta
     2041 gagtgtacaa gggcggtggc ggatctggcg gcggtggtag cgtcgacttt cagaaagcta
     2101 gctctattgt ttataagaag gagggggaac aggtggagtt cagtttccca ttggccttca
     2161 cagtcgagaa gctcacagga tcaggagagt tgtggtggca ggccgaaaga gcctcctcca
     2221 gcaagagttg gatcactttt gatctgaaaa acaaagaagt gagcgtgaag agagtcacac
     2281 aggaccctaa gctgcagatg ggcaagaagt tgcctttgca cttgacactc ccccaagccc
     2341 tgccccagta tgctggctcc gggaacctca ccctcgcctt ggaggctaag accggaaagt
     2401 tgcatcagga ggtcaacctc gtggtgatga gggcaacaca gctgcagaag aacctgacct
     2461 gcgaagtgtg gggtcccacc tcacctaaac tgatgctgtc tctgaagctg gagaataagg
     2521 aggctaaagt cagtaaaaga gagaaggctg tgtgggtcct taacccggag gcagggatgt
     2581 ggcagtgtct tcttagcgac tctggacagg tgctgctgga gagcaacatc aaggtgcttc
     2641 ctacttggtc aacaccagta cagccgatgg ccctgatcgt actcggggga gtcgccggcc
     2701 tgcttctgtt tatcggcctg ggcatcttct tctgcgtacg ctacaagtat cgcaacaggg
     2761 atgagggatc ttaccacgtt gacgaatcta ggaactatat cagtaacagc gctcagagta
     2821 atggcgccgt ggttaaagag aaacagccct ccagcgccaa atcagcaaac aaaaacaaga
     2881 aaaacaagga taaagagtac tacgtataat ctagagtcga cccgggcggc cgcttcgagc
     2941 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     3001 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     3061 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg
     3121 ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatcttcc
     3181 tagagcatgg ctacgtagat aagtagcatg gcgggttaat cattaactac aaggaacccc
     3241 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     3301 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca
     3361 gccttaatta acctaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     3421 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     3481 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg
     3541 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3601 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3661 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     3721 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     3781 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     3841 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     3901 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     3961 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
     4021 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4081 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4141 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4201 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4261 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4321 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4381 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4441 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4501 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4561 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4621 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     4681 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     4741 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     4801 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     4861 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     4921 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     4981 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     5041 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5101 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5161 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5221 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5281 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5341 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5401 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5461 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5521 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5581 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5641 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5701 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5761 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     5821 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     5881 ccgaacgacc gagcgcagcg agtcagtgag cgag
//
