LOCUS       sequence-334963-        7002 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses bioluminescent glutamate indicator in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7002)
  AUTHORS   Petersen ED, Lapan AP, Castellanos Franco EA, Fillion AJ, Crespo EL,
            Lambert GG, Grady CJ, Zanca AT, Orcutt R, Hochgeschwender U, Shaner 
            NC, Gilad AA
  TITLE     Bioluminescent Genetically Encoded Glutamate Indicators for 
            Molecular Imaging of Neuronal Activity.
  JOURNAL   ACS Synth Biol. 2023 Jul 14. doi: 10.1021/acssynbio.2c00687.
  PUBMED    37450884
REFERENCE   2  (bases 1 to 7002)
  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: "ACS Synth 
            Biol. 2023 Jul 14. doi: 10.1021/acssynbio.2c00687."
COMMENT     Sequence Label: BLING 1
FEATURES             Location/Qualifiers
     source          1..7002
                     /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"
     sig_peptide     911..970
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             2318..2464
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             2500..2541
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2551..2568
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2591..2608)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2597..2821
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2867..3295
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2954..2973)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3164..3185
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3304..3324)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3309..3638
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3489..3624
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3551..3570
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3705..4499
                     /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(3759..3778)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4369..4388
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4675..4796
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4712..4731)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4766..4785
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4845..4861)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4845..4861)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4858..4880)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4869..4885
                     /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(4893..4899,4900..4917,4918..4923))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4938..4959
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5076..5093)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5247..5835)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5327..5346)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6006..6797,6798..6866))
                     /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     6629..6648
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6867..6971)
                     /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 gctggctagt
      901 taagcttgcc atggagacag acacactcct gctatgggta ctgctgctct gggttccagg
      961 ttccactggt gacatggtct tcacactcga agatttcgtt ggggactggc gacagacagc
     1021 cggctacaac ctggaccaag tccttgaaca gggaggtgtg tccagtttgt ttcagaatct
     1081 cggggtgtcc gtaactccga tccaaaggat tgtcctgagc ggtgaaaatg ggctgaagat
     1141 cgacatccat gtcatcatcc cgtatgaagg ttctccttct agtactctgg acaaaatcgc
     1201 caaaaacggt gtgattgtcg tcggtcaccg tgaatcttca gtgcctttct cttattacga
     1261 caatcagcaa aaagtggtgg gttactcgca ggattactcc aacgccattg ttgaagcagt
     1321 gaaaaagaaa ctcaacaaac cggacttgca ggtaaaactg attccgatta caacacaaaa
     1381 ccgtattcca ctgctgcaaa acggcacttt cgattttgaa tgtggttcta ccaccaacaa
     1441 cgtcgaacgc caaaaacagg cggctttctc tgacactatt ttcgtggtcg gtacgcgcct
     1501 gttgaccaaa aagggtggcg atatcaaaga ttttgccaac ctgaaagaca aagccgtagt
     1561 cgtcacttcc ggcactacct ctgaagtttt gctcaacaaa ctgaatgaag agcaaaaaat
     1621 gaatatgcgc atcatcagcg ccaaagatca cggtgactct ttccgcaccc tggaaagcgg
     1681 tcgtgccgtt gcctttatga tggatgacgc tctgctggcc ggtgaacgtg cgaaagcgaa
     1741 gaaaccagac aactgggaaa tcgtcggcaa gccgcagtct caggaggcct acggttgtat
     1801 gttgcgtaaa gatgatccgc agttcaaaaa gctgatggat gacaccatcg ctcaggtgca
     1861 gacctccggt gaagcggaaa aatggtttga taagtggttc aaaaatccaa ttccgccgaa
     1921 aaacctgaac atgaatttcg aactgtcaga cgaaatgaaa gcactgttca aagaaccgaa
     1981 tgaccctgct cctggcctga gcggcgacca aatgggccag atcgaaaaaa tttttaaggt
     2041 ggtgtaccct gtggatgatc atcactttaa ggtgatcctg cactatggca cactggtaat
     2101 cgacggggtt acgccgaaca tgatcgacta tttcggacgg ccgtatgaag gcatcgccgt
     2161 gttcgacggc aaaaagatca ctgtaacagg gaccctgtgg aacggcaaca aaattatcga
     2221 cgagcgcctg atcaaccccg acggctccct gctgttccga gtaaccatca acggagtgac
     2281 cggctggcgg ctgtgcgaac gcattctggc gctcgaggct gtgggccagg acacgcagga
     2341 ggtcatcgtg gtgccacact ccttgccctt taaggtggtg gtgatctcag ccatcctggc
     2401 cctggtggtg ctcaccatca tctcccttat catcctcatc atgctttggc agaagaagcc
     2461 acgttagtaa catatgctct agagggcccg cggttcgaag gtaagcctat ccctaaccct
     2521 ctcctcggtc tcgattctac gcgtaccggt catcatcacc atcaccattg agtttaaacc
     2581 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     2641 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     2701 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     2761 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     2821 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     2881 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2941 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3001 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3061 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     3121 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3181 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataatg gattttgccg
     3241 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     3301 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3361 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3421 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3481 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3541 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     3601 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     3661 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     3721 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     3781 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     3841 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     3901 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     3961 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4021 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4081 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4141 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4201 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     4261 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4321 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4381 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4441 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     4501 cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     4561 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     4621 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     4681 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     4741 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     4801 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     4861 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     4921 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     4981 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     5041 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5101 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5161 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5221 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5281 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5341 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5401 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5461 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5521 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5581 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5641 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     5701 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5761 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5821 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5881 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5941 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6001 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6061 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6121 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6181 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6241 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6301 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6361 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6421 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6481 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6541 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6601 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6661 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6721 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6781 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6841 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     6901 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6961 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
