LOCUS       sequence-338886-        6554 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BRET-Based Bioluminescent Red Protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6554)
  AUTHORS   Tian X, Zhang Y, Li X, Xiong Y, Wu T, Ai HW
  TITLE     A luciferase prosubstrate and a red bioluminescent calcium indicator
            for imaging neuronal activity in mice.
  JOURNAL   Nat Commun. 2022 Jul 8;13(1):3967. doi: 10.1038/s41467-022-31673-x.
  PUBMED    35803917
REFERENCE   2  (bases 1 to 6554)
  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"; journalName: "Nat 
            Commun."; date: "2022-07-8"; volume: "13(1):3967. doi"; pages: " 
            10.1038/s41467-022-31673-x"
COMMENT     Sequence Label: pcDNA3-BREP
FEATURES             Location/Qualifiers
     source          1..6554
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      895..904
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1034..1052)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1302..1321
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        complement(2107..2125)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2108..2125)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2145..2162)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2151..2375
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2421..2849
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2508..2527)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2718..2739
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2858..2878)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2863..3192
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3043..3178
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3105..3124
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3259..4053
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3313..3332)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3923..3942
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4227..4348
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4264..4283)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4318..4337
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4397..4413)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4397..4413)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4410..4432)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4421..4437
                     /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(4445..4451,4452..4469,4470..4475))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4490..4511
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4628..4645)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4799..5387)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4879..4898)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5558..6349,6350..6418))
                     /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     6181..6200
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6419..6523)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgccacc
      901 atggtgagca agggcgaggc agtgatcaag gagttcatgc ggttcaaggt gcacatggag
      961 ggctccatga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
     1021 ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt ctcctgggac
     1081 atcctgtccc ctcagttcat gtacggctcc agggccttca tcaagcaccc cgccgacatc
     1141 cccgactact ataagcagtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
     1201 gaggacggcg gcgccgtgac cgtgacccag gacacctccc tggaggacgg caccctgatc
     1261 tacaaggtga agctccgcgg caccaacttc cctcctgacg gccccgtaat gcagaagaag
     1321 acaatgggct gggaagcgtc caccgagcgg ttgtaccccg aggacggcgt gctgaagggc
     1381 gacattaaga tggccctgcg cctgaaggac ggcggccgct acctggcgga cttcaagacc
     1441 acctacaagg ccaagaagcc cgtgcagatg cccggcgcct acaacgtcga ccgcaagttg
     1501 gacatcacct cccacaacga ggactacacc gtggtggaac agtacgaacg ctccgagggc
     1561 cgccatcttg atacactcga agatttcgtt ggggactggc gacagacagc cggctacaac
     1621 ttgagtcaag tccttgaaca gggaggtgtg tccagtttgt ttcagaatct cggggtgtcc
     1681 gtaactccga tccaaaggat tgtcctgagc ggtgaaaatg ggctgaagat cgacatccat
     1741 gtcatcatcc cgtatgaagg tctgagcggc gaccaaatgg gccagatcga aaaaattttt
     1801 aaggtggtgt accctgtgga taatcatcac tttaaggtga tcctgcacta tggcacactg
     1861 gtaatcgacg gggttacgcc gaacatgatc gactatttcg gacggccgta tgaaggcatc
     1921 gccgtgttcg acggcaaaaa gatcactgta acagggaccc tgtggaacgg caacaaaatt
     1981 atcgacgagc gcctgatcaa ccccgacggc tccctgctgt tccgagtaac catcaacgga
     2041 gtgaccggct ggcgtctgca tgaacgcatt ctggcgtaac tcgagcatgc atctagaggg
     2101 ccctattcta tagtgtcacc taaatgctag agctcgctga tcagcctcga ctgtgccttc
     2161 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     2221 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     2281 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     2341 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     2401 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2461 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2521 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2581 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     2641 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2701 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     2761 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     2821 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     2881 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     2941 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3001 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3061 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3121 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3181 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     3241 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     3301 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     3361 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     3421 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     3481 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     3541 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     3601 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     3661 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     3721 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     3781 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     3841 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     3901 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     3961 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     4021 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     4081 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     4141 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     4201 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     4261 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4321 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     4381 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     4441 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     4501 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4561 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     4621 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4681 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4741 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4801 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4861 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4921 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4981 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5041 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5101 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5161 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5221 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5281 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5341 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5401 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5461 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5521 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5581 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5641 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5701 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5761 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5821 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     5881 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5941 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6001 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6061 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6121 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6181 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6241 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6301 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6361 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6421 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6481 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6541 tgccacctga cgtc
//
