LOCUS       sequence-344153-        6549 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  genetically encoded GRP sensor grpLight1.2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6549)
  AUTHORS   Melzer S, Newmark ER, Mizuno GO, Hyun M, Philson AC, Quiroli E, 
            Righetti B, Gregory MR, Huang KW, Levasseur J, Tian L, Sabatini BL
  TITLE     Bombesin-like peptide recruits disinhibitory cortical circuits and 
            enhances fear memories.
  JOURNAL   Cell. 2021 Oct 28;184(22):5622-5634.e25. doi: 
            10.1016/j.cell.2021.09.013. Epub 2021 Oct 4.
  PUBMED    34610277
REFERENCE   2  (bases 1 to 6549)
  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.cell.2021.09.013"; journalName: "Cell"; date: 
            "2021-10-28- 28"; volume: "184"; issue: "22"; pages: "5622-5634.e25"
COMMENT     Sequence Label: pAAV hSyn grpLight1.2
FEATURES             Location/Qualifiers
     source          1..6549
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   13..142
                     /label=AAV2 ITR
     promoter        224..671
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     misc_feature    2675..3263
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2728..2748)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3146..3157)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3266..3282)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3267..3283)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3312..3433
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3323..3342)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3377..3396
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3595..3724
                     /label=AAV2 ITR
     primer_bind     complement(3844..3864)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3844..3862)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3883..3899)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3883..3899)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3896..3918)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3907..3923
                     /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(3931..3937,3938..3955,3956..3961))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3976..3997
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4285..4873)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4365..4384)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5044..5835,5836..5904))
                     /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     5667..5686
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5905..6009)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6035..6490)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6172..6193)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6384..6403
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctattacgcc agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt
       61 cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
      121 aactccatca ctaggggttc cttgtagtta atgattaacc cgccatgcta cttatctacg
      181 tagccatgct ctaggaagat ctctgcagag ggccctgcgt atgagtgcaa gtgggtttta
      241 ggaccaggat gaggcggggt gggggtgcct acctgacgac cgaccccgac ccactggaca
      301 agcacccaac ccccattccc caaattgcgc atcccctatc agagaggggg aggggaaaca
      361 ggatgcggcg aggcgcgtgc gcactgccag cttcagcacc gcggacagtg ccttcgcccc
      421 cgcctggcgg cgcgcgccac cgccgcctca gcactgaagg cgcgctgacg tcactcgccg
      481 gtcccccgca aactcccctt cccggccacc ttggtcgcgt ccgcgccgcc gccggcccag
      541 ccggaccgca ccacgcgagg cgcgagatag gggggcacgg gcgcgaccat ctgcgctgcg
      601 gcgccggcga ctcagcgctg cctcagtctg cggtgggcag cggaggagtc gtgtcgtgcc
      661 tgagagcgca gtcgaattca agctgctagc aaggatccac ccgccagcca tgaagacgat
      721 catcgccctg agctacatct tctgcctggt gttcgccgac tacaaggacg atgatgacgc
      781 catggcgctg aatgattgct ttctgctcaa cttggaggtg gatcatttta tgcactgtaa
      841 catcagttcc cactccgcag atcttcccgt gaatgacgac tggtcacacc caggaattct
      901 ctatgtaata ccagcagtgt atggcgtcat catactgatc ggtctcatag gaaatattac
      961 gcttatcaag atcttttgca ccgtaaagtc catgagaaac gtccccaacc tttttattag
     1021 cagcctcgca cttggggatc tgctgcttct tataacttgt gctccagtag atgcaagtag
     1081 gtacttggcg gacagatggc tgtttggtag aatagggtgc aaacttattc ctttcataca
     1141 gctcacgagc gtcggcgttt ctgtttttac cctcacagcg ctgagcgctg atcggtataa
     1201 agcaatcgtc cgcccgaagg acatccaagc ctctcatgcg cttatgaaga tatgccttaa
     1261 agccgctttc atttggataa tatccatgct ccttgcgatt ccggaagctg ttttctctga
     1321 tctccacccc tttcatgaag agagcacaaa tcaaaccttt atcagttgcg cgccgtaccc
     1381 ccacagtaat gagctccatc ccaagatcca ctctatggcg agcttccttg tcttttatgt
     1441 aattccgctt tctataattt ctgtctatta ctactttatc gctaaaaacc agtctgccta
     1501 tctcagcagc ggcataaacg tatatattaa ggctgacaaa caaaaaaacg gtatcaaagc
     1561 aaacttcaag ataagacata acatagagga tggaggagtc caattggctt accattacca
     1621 gcaaaatacc ccaatcggtg acggcccagt gctgcttccg gataaccact atctcagcgt
     1681 acaaagcaaa cttagtaaag atccgaacga aaaaagggac cacatggtcc tgttggagtt
     1741 cgtcacagcc gccgggatta ctttggggat ggacgagttg tacaaaggtg gtaccggggg
     1801 aagcatggta agcaagggtg aagaactctt tacgggcgta gtaccaattc ttgtagagtt
     1861 ggacggagac gttaacggtc ataaattctc agtcagcggc gagggcgaag gagacgcgac
     1921 ttacggtaaa cttacgctca agtttatttg cacgaccggg aagctgccag tgccttggcc
     1981 cacactggtg actactttga cctatggtgt acaatgtttt tcaagatacc ctgaccatat
     2041 gaaacaacac gactttttca agagcgcgat gccagaggga tatattcagg aacggaccat
     2101 cttcttcaaa gatgacggca actataaaac gagggcggaa gtgaagttcg agggcgatac
     2161 actcgtgaat aggatcgaac tcaagggaat agattttaag gaagacggga atatcttggg
     2221 gcataagctg gagtacaatc cccacgacca actgaaacaa atagaatcca ggaaacggtt
     2281 ggcgaaaact gtactggttt ttgtagggct ttttgcattt tgctggcttc cgaatcacgt
     2341 tatctacctg taccgcagct accattatag cgaggtggat acgtcaatgc ttcactttgt
     2401 tacctcaatt tgcgctcggc tgctggcctt tactaactca tgcgttaatc cgttcgccct
     2461 gtatcttctg tctaaatcat ttcgcaaaca gtttaacacc cagctcctct gttgtcagcc
     2521 cggccttatt atacgcagcc actaaaccgg caggtctacc acttgcatga ctagcttgaa
     2581 atcaacaaat ccgagtgtgg ccacattttc ccttataaac ggcaacattt gtcacgaacg
     2641 ctacgtgtga tgtacagcgg ccaagcttat cgataatcaa cctctggatt acaaaatttg
     2701 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     2761 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     2821 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     2881 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     2941 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     3001 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     3061 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     3121 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     3181 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     3241 ctccctttgg gccgcctccc cgcatcgata ccgtcgacct cgacccgggc ggccgcttcg
     3301 agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     3361 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     3421 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat
     3481 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatct
     3541 tcctagagca tggctacgta gataagtagc atggcgggtt aatcattaac tacaaggaac
     3601 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc
     3661 gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc
     3721 gcagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgtc atcgttacca
     3781 tgttgcaaga ctacaagagt attgcataag actacattaa gcttgcagct ccagcttttg
     3841 ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt
     3901 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     3961 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     4021 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     4081 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4141 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4201 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4261 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4321 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4381 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4441 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4501 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4561 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4621 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4681 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4741 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4801 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4861 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4921 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4981 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5041 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5101 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5161 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     5221 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     5281 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     5341 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5401 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5461 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5521 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5581 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5641 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5701 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5761 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5821 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5881 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     5941 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6001 ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta
     6061 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     6121 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     6181 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     6241 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     6301 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     6361 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     6421 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     6481 cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     6541 gcctcttcg
//
