LOCUS       sequence-501844-        6687 bp    DNA     circular SYN 04-MAR-2026
DEFINITION  Expression of Cre-dependent binding mutant control calcium lifetime 
            sensor LifeCamp0 under hSyn promoter in pAAV backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6687)
  AUTHORS   Lodder B, Raghubardayal M, Ganesh S, Cai X, Stern J, Sherman M, 
            Rosen P, Kamath T, Hartman I, Siegel M, Timmins J, Adan R, Andermann
            M, Sabatini BL
  TITLE     Rapid fluorescence lifetime sensor development of LifeCamp enables 
            transient and baseline absolute calcium measurements
  JOURNAL   bioRxiv 2025.12.23.696288
REFERENCE   2  (bases 1 to 6687)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 4, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2025.12.23.696288"
COMMENT     Sequence Label: pAAV-hsyn-DIO-LifeCamp0
FEATURES             Location/Qualifiers
     source          1..6687
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    21..54
                     /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    117..150
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     1563..1582
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1822..1843
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1949..1970)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(2240..2257)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(2277..2310)
                     /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(2381..2414)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2447..3035
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2500..2520)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2918..2929)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3038..3054)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3039..3055)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3084..3205
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3095..3114)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3149..3168
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3367..3496
                     /label=AAV2 ITR
     rep_origin      complement(3733..4321)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3813..3832)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4492..5283,5284..5352))
                     /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     5115..5134
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5353..5457)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5483..5938)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5620..5641)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5832..5851
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   6010..6139
                     /label=AAV2 ITR
     promoter        6221..6668
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
ORIGIN
        1 tgcagagata cggtgcattt ataacttcgt ataaagtatc ctatacgaag ttatatcaaa
       61 ataggaagac caatgcttca ccatcgaccc gaattgccaa gcatcaccat cgacccataa
      121 cttcgtataa tgtatgctat acgaagttat actagagctt tcatttatag agttcgtcca
      181 ttcccaatgt aataccggct gcggtaacaa attccaagag aaccatatga tctctctttt
      241 cgtttgggtc cttagagagt gcgctctgtg ttgagagata atgattgtca ggcagcaaga
      301 cgggtccgtc cccgattgga gtgttctgtt ggtagtggtc ggcaagctga acactcccgt
      361 cttcaatatt gtgcctgatc ttgtagttta ctttgattcc gttcttctgt ttatctgcca
      421 ttatataaac attatggctg ttcatgttat actcaagctt atgccccaaa atgttcccgt
      481 cttctttaaa atctatcccc ttcaattcta tgcgattgac aagggtgtcg ccctcgaatt
      541 tgacctctgc tcgagtctta taattaccat cgtctttaaa gaatatggtt cgttcttgca
      601 cataaccttc aggcatggcg ctcttaaaga agtcatgctg cttcatatga tctggatacc
      661 gactgaagca ctgcactcca tatccaagag tagtgacaag tgtaggccaa ggaacgggca
      721 atttcccggt ggtacagata aatttaagag taagtttgcc atatgttgca tccccttcgc
      781 cttcgccaga aacagagaat ttgtgaccgt taacgtcgcc atcgagctcg acaagtatgg
      841 ggacaactcc tgtgaagagt tcttcccctt tggataccat ggacccacct gatcccccgg
      901 acccgccgct gccaccactt cctccggaac caccgctccc tccggagccc ccgcttcccc
      961 cactacctcc agagccccct gatccaccag atcctccaga gccgcccttc gctgtcatca
     1021 tttgtacaaa ctcttcgtag tttacctgac catccccatc gatgtctgct tccctgatca
     1081 tttcatcaac ctcttcatct gttaacttct ctccaaggtt tgtcatcacg tggcgaagct
     1141 ctgctgcact gatgtagcca ttgccatcct tatcaaacac accgaacgct tctctaattt
     1201 cttcttccgt gtccctgtat ttcatttttc ttgccatcat tgtcaggaac tcagggaagt
     1261 cgattgtgcc gtcaccgtcg gcatctactt cattgatcat gtcctgcagc tctgcttctg
     1321 tggggttctg ccccagagac cgcatcaccg tccccagctc cttggttgtt attgtcccat
     1381 ccccgtcctt gtcaaatagg gagaaagcct ctttgtattc tgcgatctgc tcttcagtca
     1441 gttggtccgg caggttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt
     1501 cgatgccctt cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc
     1561 gggtcttgta gttgccgtcg tccttgaaga agatggtgcg ctcctggatg tagccttcgg
     1621 gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg ctgaagcact
     1681 gcacgccgta ggtcagggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg
     1741 tgcagatgaa cttcagggtc agcttgccgt aggtggcatc gccctcaccc tcgccggaca
     1801 cgctgaactt gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg
     1861 tgaacagctc ctcgcccttg ctcaccatgc tccctccggt accgcccttg tacagctcgt
     1921 ccatgccgag agtgatcccg gcggcggtca cgaactccag caggaccatg tgatcgcgct
     1981 tctcgttggg gtctttcgaa agtttggact ccacgctcag gtagtggttg tcgggcagca
     2041 gcacggggcc gtcgccgatg ggggtgttct gctggtagtg gtaggcgagc tgcacgccgc
     2101 cgtcctcgat gttgtggcgg atgtggaagt tcgccttgat gccgttcttc tgcttgtcgg
     2161 ccttgatata gacgttgatc ttcagtcctc cagatcctcc agatcctcca gatcctccag
     2221 atcctccaga tcctcccgtg tgatgatggt gatgatgcat ggtggcggat cctagtataa
     2281 cttcgtatag gatactttat acgaagttat cattgggatt cttcctattt tgatccaagc
     2341 atcaccatcg accctctagt ccagatctca ccatcgaccc ataacttcgt atagcataca
     2401 ttatacgaag ttatatctaa acgtgtcacg agtgaagctt atcgataatc aacctctgga
     2461 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     2521 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     2581 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     2641 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     2701 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     2761 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     2821 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
     2881 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     2941 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     3001 gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgtcgac ctcgacccgg
     3061 gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag
     3121 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     3181 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     3241 tcagggggag atgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaaaat
     3301 cgataaggat cttcctagag catggctacg tagataagta gcatggcggg ttaatcatta
     3361 actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     3421 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     3481 gcgagcgagc gcgcagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3541 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3601 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3661 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3721 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3781 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3841 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3901 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3961 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4021 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4081 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4141 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4201 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4261 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4321 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4381 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4441 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4501 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4561 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4621 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4681 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     4741 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4801 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4861 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4921 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     4981 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5041 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5101 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5161 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5221 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5281 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5341 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5401 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5461 atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta
     5521 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     5581 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     5641 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     5701 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     5761 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     5821 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     5881 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc
     5941 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     6001 ttacgccagc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc cgggcgtcgg
     6061 gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac
     6121 tccatcacta ggggttcctt gtagttaatg attaacccgc catgctactt atctacgtag
     6181 ccatgctcta ggaagatctc tgcagagggc cctgcgtatg agtgcaagtg ggttttagga
     6241 ccaggatgag gcggggtggg ggtgcctacc tgacgaccga ccccgaccca ctggacaagc
     6301 acccaacccc cattccccaa attgcgcatc ccctatcaga gagggggagg ggaaacagga
     6361 tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg gacagtgcct tcgcccccgc
     6421 ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc gctgacgtca ctcgccggtc
     6481 ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg cgccgccgcc ggcccagccg
     6541 gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg cgaccatctg cgctgcggcg
     6601 ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg aggagtcgtg tcgtgcctga
     6661 gagcgcagtc gaattcaagc tgctagc
//
