LOCUS       sequence-501847-        6699 bp    DNA     circular SYN 04-MAR-2026
DEFINITION  Expression of Cre-dependent calcium lifetime sensor LifeCamp under 
            hSyn promoter in pAAV backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6699)
  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 6699)
  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-LifeCamp
FEATURES             Location/Qualifiers
     source          1..6699
                     /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(2252..2269)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(2289..2322)
                     /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(2393..2426)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2459..3047
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2512..2532)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2930..2941)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3050..3066)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3051..3067)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3096..3217
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3107..3126)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3161..3180
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3379..3508
                     /label=AAV2 ITR
     rep_origin      complement(3745..4333)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3825..3844)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4504..5295,5296..5364))
                     /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     5127..5146
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5365..5469)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5495..5950)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5632..5653)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5844..5863
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   6022..6151
                     /label=AAV2 ITR
     promoter        6233..6680
                     /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 ttcaggccca tcaggcggcc gatgatcttc acggcggtgg
     2221 ccacctcctt gaaggtcttc ttgcgacgcg tgtgatgatg gtgatgatgc atggtggcgg
     2281 atcctagtat aacttcgtat aggatacttt atacgaagtt atcattggga ttcttcctat
     2341 tttgatccaa gcatcaccat cgaccctcta gtccagatct caccatcgac ccataacttc
     2401 gtatagcata cattatacga agttatatct aaacgtgtca cgagtgaagc ttatcgataa
     2461 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     2521 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     2581 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     2641 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     2701 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     2761 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     2821 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     2881 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     2941 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     3001 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgtcg
     3061 acctcgaccc gggcggccgc ttcgagcaga catgataaga tacattgatg agtttggaca
     3121 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     3181 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     3241 tatgtttcag gttcaggggg agatgtggga ggttttttaa agcaagtaaa acctctacaa
     3301 atgtggtaaa atcgataagg atcttcctag agcatggcta cgtagataag tagcatggcg
     3361 ggttaatcat taactacaag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     3421 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     3481 cggcctcagt gagcgagcga gcgcgcagct gcattaatga atcggccaac gcgcggggag
     3541 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     3601 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3661 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3721 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3781 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3841 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3901 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3961 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4021 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4081 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4141 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4201 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4261 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4321 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4381 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4441 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4501 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4561 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4621 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4681 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4741 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4801 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4861 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4921 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     4981 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5041 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5101 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5161 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5221 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5281 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5341 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     5401 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     5461 ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta
     5521 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     5581 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     5641 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     5701 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     5761 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     5821 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     5881 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     5941 cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     6001 gcctcttcgc tattacgcca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag
     6061 cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag
     6121 ggagtggcca actccatcac taggggttcc ttgtagttaa tgattaaccc gccatgctac
     6181 ttatctacgt agccatgctc taggaagatc tctgcagagg gccctgcgta tgagtgcaag
     6241 tgggttttag gaccaggatg aggcggggtg ggggtgccta cctgacgacc gaccccgacc
     6301 cactggacaa gcacccaacc cccattcccc aaattgcgca tcccctatca gagaggggga
     6361 ggggaaacag gatgcggcga ggcgcgtgcg cactgccagc ttcagcaccg cggacagtgc
     6421 cttcgccccc gcctggcggc gcgcgccacc gccgcctcag cactgaaggc gcgctgacgt
     6481 cactcgccgg tcccccgcaa actccccttc ccggccacct tggtcgcgtc cgcgccgccg
     6541 ccggcccagc cggaccgcac cacgcgaggc gcgagatagg ggggcacggg cgcgaccatc
     6601 tgcgctgcgg cgccggcgac tcagcgctgc ctcagtctgc ggtgggcagc ggaggagtcg
     6661 tgtcgtgcct gagagcgcag tcgaattcaa gctgctagc
//
