LOCUS       sequence-501848-        6956 bp    DNA     circular SYN 04-MAR-2026
DEFINITION  Expression of calcium lifetime sensor LifeCamp under CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6956)
  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 6956)
  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: CAG-LifeCamp
FEATURES             Location/Qualifiers
     source          1..6956
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    828..962
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(852..871)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     906..925
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(968..1163)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      982..1117
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    1222..1243
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1258..1263,1264..1281,1282..1288)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1296..1312
                     /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     1301..1323
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1320..1336
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1320..1336
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1679..1698
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    1697..1752
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     1798..1817
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     primer_bind     3325..3344
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3584..3605
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3711..3732)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(4014..4031)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4088..4107)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          4115..5131
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(4169..4191)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(5132..5409)
                     /label=chicken beta-actin promoter
     enhancer        5411..5790
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5821..5925
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5926..5994,5995..6786)
                     /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(6144..6163)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagcggat ccagacatga
      841 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
      901 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
      961 ttagcttttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct
     1021 cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg
     1081 gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta
     1141 tggttgctga ctaattgaga tgcggatccg ctggcacgac aggtttcccg actggaaagc
     1201 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     1261 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     1321 aggaaacagc tatgaccatg attacgccaa gcttgggctg caggtcgagg gatctccata
     1381 agagaagagg gacagctatg actgggagta gtcaggagag gaggaaaaat ctggctagta
     1441 aaacatgtaa ggaaaatttt agggatgtta aagaaaaaaa taacacaaaa caaaatataa
     1501 aaaaaatcta acctcaagtc aaggcttttc tatggaataa ggaatggaca gcagggggct
     1561 gtttcatata ctgatgacct ctttatagcc aacctttgtt catggcagcc agcatatggg
     1621 catatgttgc caaactctaa accaaatact cattctgatg ttttaaatga tttgccctcc
     1681 catatgtcct tccgagtgag agacacaaaa aattccaaca cactattgca atgaaaataa
     1741 atttccttta ttagccagaa gtcagatgct caaggggctt catgatgtcc ccataatttt
     1801 tggcagaggg aaaaagatct cagtggtatt tgtgagccag ggcattggcc acaccagcca
     1861 ccaccttctg ataggcagcc tgcacctgag gagtgcggcc gccactcgtg acacgtttag
     1921 attcatttat agagttcgtc cattcccaat gtaataccgg ctgcggtaac aaattccaag
     1981 agaaccatat gatctctctt ttcgtttggg tccttagaga gtgcgctctg tgttgagaga
     2041 taatgattgt caggcagcaa gacgggtccg tccccgattg gagtgttctg ttggtagtgg
     2101 tcggcaagct gaacactccc gtcttcaata ttgtgcctga tcttgtagtt tactttgatt
     2161 ccgttcttct gtttatctgc cattatataa acattatggc tgttcatgtt atactcaagc
     2221 ttatgcccca aaatgttccc gtcttcttta aaatctatcc ccttcaattc tatgcgattg
     2281 acaagggtgt cgccctcgaa tttgacctct gctcgagtct tataattacc atcgtcttta
     2341 aagaatatgg ttcgttcttg cacataacct tcaggcatgg cgctcttaaa gaagtcatgc
     2401 tgcttcatat gatctggata ccgactgaag cactgcactc catatccaag agtagtgaca
     2461 agtgtaggcc aaggaacggg caatttcccg gtggtacaga taaatttaag agtaagtttg
     2521 ccatatgttg catccccttc gccttcgcca gaaacagaga atttgtgacc gttaacgtcg
     2581 ccatcgagct cgacaagtat ggggacaact cctgtgaaga gttcttcccc tttggatacc
     2641 atggacccac ctgatccccc ggacccgccg ctgccaccac ttcctccgga accaccgctc
     2701 cctccggagc ccccgcttcc cccactacct ccagagcccc ctgatccacc agatcctcca
     2761 gagccgccct tcgctgtcat catttgtaca aactcttcgt agtttacctg accatcccca
     2821 tcgatgtctg cttccctgat catttcatca acctcttcat ctgttaactt ctctccaagg
     2881 tttgtcatca cgtggcgaag ctctgctgca ctgatgtagc cattgccatc cttatcaaac
     2941 acaccgaacg cttctctaat ttcttcttcc gtgtccctgt atttcatttt tcttgccatc
     3001 attgtcagga actcagggaa gtcgattgtg ccgtcaccgt cggcatctac ttcattgatc
     3061 atgtcctgca gctctgcttc tgtggggttc tgccccagag accgcatcac cgtccccagc
     3121 tccttggttg ttattgtccc atccccgtcc ttgtcaaata gggagaaagc ctctttgtat
     3181 tctgcgatct gctcttcagt cagttggtcc ggcaggttgt actccagctt gtgccccagg
     3241 atgttgccgt cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg
     3301 ccctcgaact tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg
     3361 cgctcctgga tgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg
     3421 tcggggtagc ggctgaagca ctgcacgccg taggtcaggg tggtcacgag ggtgggccag
     3481 ggcacgggca gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca
     3541 tcgccctcac cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg
     3601 accaggatgg gcaccacccc ggtgaacagc tcctcgccct tgctcaccat gctccctccg
     3661 gtaccgccct tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc
     3721 agcaggacca tgtgatcgcg cttctcgttg gggtctttcg aaagtttgga ctccacgctc
     3781 aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag
     3841 tggtaggcga gctgcacgcc gccgtcctcg atgttgtggc ggatgtggaa gttcgccttg
     3901 atgccgttct tctgcttgtc ggccttgata tagacgttga tcttcaggcc catcaggcgg
     3961 ccgatgatct tcacggcggt ggccacctcc ttgaaggtct tcttgcgacg cgtgtgatga
     4021 tggtgatgat gcatggtggc aaatgcaccg tatctctgca gaattctttg ccaaaatgat
     4081 gagacagcac aataaccagc acgttgccca ggagctgtag gaaaaagaag aaggcatgaa
     4141 catggttagc agaggctcta gagccgccgg tcacacgcca gaagccgaac cccgccctgc
     4201 cccgtccccc ccgaaggcag ccgtcccccc gcggacagcc ccgaggctgg agagggagaa
     4261 ggggacggcg gcgcggcgac gcacgaaggc cctccccgcc catttccttc ctgccggcgc
     4321 cgcaccgctt cgccccgcgc ccgctagagg gggtgcggcg gcgcctccca gatttcggct
     4381 ccgcacagat ttgggacaaa ggaagtccct gcgccctctc gcacgattac cataaaaggc
     4441 aatggctgcg gctcgccgcg cctcgacagc cgccggcgct ccgggggccg ccgcgcccct
     4501 cccccgagcc ctccccggcc cgaggcggcc ccgccccgcc cggcaccccc acctgccgcc
     4561 accccccgcc cggcacggcg agccccgcgc cacgccccgt acggagcccc gcacccgaag
     4621 ccgggccgtg ctcagcaact cggggagggg ggtgcagggg gggttgcagc ccgaccgacg
     4681 cgcccacacc ccctgctcac ccccccacgc acacaccccg cacgcagcct ttgttcccct
     4741 cgcagccccc cccgcaccgc ggggcaccgc ccccggccgc gctcccctcg cgcacactgc
     4801 ggagcgcaca aagccccgcg ccgcgcccgc agcgctcaca gccgccgggc agcgcggagc
     4861 cgcacgcggc gctccccacg cacacacaca cgcacgcacc ccccgagccg ctccccccgc
     4921 acaaagggcc ctcccggagc ccctcaaggc tttcacgcag ccacagaaaa gaaacaagcc
     4981 gtcattaaac caagcgctaa ttacagcccg gaggagaagg gccgtcccgc ccgctcacct
     5041 gtgggagtaa cgcggtcagt cagagccggg gcgggcggcg cgaggcggcg gcggagcggg
     5101 gcacggggcg aaggcagcgc gcagcgactc ccgcccgccg cgcgcttcgc tttttatagg
     5161 gccgccgccg ccgccgcctc gccataaaag gaaactttcg gagcgcgccg ctctgattgg
     5221 ctgccgccgc acctctccgc ctcgccccgc cccgcccctc gccccgcccc gccccgcctg
     5281 gcgcgcgccc cccccccccc cccgccccca tcgctgcaca aaataattaa aaaataaata
     5341 aatacaaaat tgggggtggg gagggggggg agatggggag agtgaagcag aacgtggggc
     5401 tcacctcgac catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt
     5461 cccataaggt catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa
     5521 tagggggcgt acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta
     5581 aatactccac ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg
     5641 tcattattga cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg
     5701 taagttatgt aacgcggaac tccatatatg ggctatgaac taatgacccc gtaattgatt
     5761 actattaata actagtcaat aatcaatgtc gacccaggtg gcacttttcg gggaaatgtg
     5821 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     5881 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     5941 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     6001 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     6061 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     6121 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     6181 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     6241 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     6301 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     6361 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     6421 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     6481 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     6541 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     6601 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     6661 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     6721 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     6781 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     6841 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6901 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttc
//
