LOCUS       sequence-364510-        7517 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of a fluorescent sensor for C3a complement in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7517)
  AUTHORS   Ino D, Tanaka Y, Hibino H, Nishiyama M
  TITLE     A fluorescent sensor for real-time measurement of extracellular 
            oxytocin dynamics in the brain.
  JOURNAL   Nat Methods. 2022 Sep 22. pii: 10.1038/s41592-022-01597-x. doi: 
            10.1038/s41592-022-01597-x.
  PUBMED    36138174
REFERENCE   2  (bases 1 to 7517)
  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 
            Methods. 2022 Sep 22. pii: 10.1038/s41592-022-01597-x. doi: 
            10.1038/s41592-022-01597-x."
COMMENT     Sequence Label: pCIS.MTRIA-C3a
FEATURES             Location/Qualifiers
     source          1..7517
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          123..1139
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1063..1085
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1147..1166
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     1195..1211
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1196..1212
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2534..2555
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2661..2682)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3412..3428)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3412..3428)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3517..3536)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3582..3637
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3636..3655)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     promoter        complement(3992..4010)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4031..4047)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4031..4047)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4044..4066)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4055..4071
                     /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(4079..4085,4086..4103,4104..4109))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4124..4145
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4262..4279)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4433..5021)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4513..4532)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5192..5983,5984..6052))
                     /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     5815..5834
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6053..6157)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6184..6639
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6271..6290)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6481..6502
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6765..6787
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6779..6796
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6780..6796
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6803..6822
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6803..6821
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        6886..7189
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
       61 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      121 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      181 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      241 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      301 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      361 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      421 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
      481 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
      541 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
      601 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
      661 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
      721 cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg
      781 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
      841 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
      901 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
      961 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1021 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1081 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1141 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagga ttcctcgagg
     1201 tcgacggtat cgataagctt gatatcgaat tcaccatgga gtctttcgat gctgacacca
     1261 attcaactga cctacactca cggcctctgt ttcaacccca agacattgcc tccatggtca
     1321 ttcttggtct cacttgtcta ttgggactgc taggcaatgg gctggtgctg tgggtagctg
     1381 gcgtaaagat gaagacgacc gtgaacacag tctggttcct ccatctcacc ctggccgatt
     1441 tcctctgctg cctctccttg cccttctcct tggctcacct gattctccaa ggacactggc
     1501 cctatggctt gttcctgtgc aaacttatcc catccatcat tattctcaac atgtttgcca
     1561 gtgtcttcct gcttactgcc attagcctgg accgatgtct gatagtacat aagccaatct
     1621 ggtgccagaa tcatcgaaac gtgagaaccg ccttcgccat ctgtggatgt gtctgggtgg
     1681 tagcctttgt gatgtgtgtg cccgtatttg tataccgtga tctgttcatt atggacaatc
     1741 gcagtatatg tagatataat tttgattcct ccaggtcata tgattattgg gactacgtgt
     1801 acaaactaag tctaccagaa agcaattcta ctgataactc cactgctcag ctaactggac
     1861 atatgaatga caggtcagct ccttcctctg tacaggcaag ggattacttt tggacagtta
     1921 ccactgccct ccagtcacag ccattcctaa catctcctga agactcattc tctctagatt
     1981 cagcaaacca acaaccccat tatggtggaa agcctcctaa tgtcctcaca gccgccgtac
     2041 ccagcgggtt tcctgttgaa gatcgtaaat ccaatacact gaacgctgac gcttttctct
     2101 ctgctcacac agaacttttc cctactgctt ctagtggtca tttatacccc tatgatttcc
     2161 agggggatta tgttgaccaa ttcacgtatg acaatcatgt gccgacaccg ctgatggcaa
     2221 taaccatcac aaggctggtg gtgggcttcc tggtgccgtt tttcatcatg gtaatttgtt
     2281 acagcctcat ctgctacaaa atatggcgaa atttcaaagg gaaattaaac gtctatatca
     2341 aggccgacaa gcagaagaac ggcatcaagg cgaacttcca catccgccac aacatcgagg
     2401 acggcggcgt gcagctcgcc taccactacc agcagaacac ccccatcggc gacggccccg
     2461 tgctgctgcc cgacaaccac tacctgagcg tgcagaccaa actttcgaaa gaccccaacg
     2521 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     2581 tggacgagct gtacaagggc ggtaccggag ggatgatggt gagaaagggc gaggagctgt
     2641 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     2701 gcgtgtccgg cgagggtgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     2761 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
     2821 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     2881 tgcccgaagg ctacatccag gagcgcacca tcttcttcaa ggacgacggc cactacaaga
     2941 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     3001 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tcagtaatct
     3061 ccaaggcaaa gatacgcaca gtgaaaatgg ctgtggctgt ggtcactgtc ttttttatct
     3121 gctggactcc ataccatctt gtcggagtcc tgctattgat tactgatcca gaaagttcct
     3181 tgggggaagc tgtgatgtcc tgggaccaca tgtccattgc tttagcatct gccaatagtt
     3241 gcttcaaccc tttcctgtat gccctcttgg ggaaagactt taggaagaaa gcaagacagt
     3301 ctataaaggg cattctggaa gcagccttca gcgaagagct cacgcactct accaactgta
     3361 cccaagacaa agcctcttca aaaagaaaca atatgagtac agatgtgtga ggatccacta
     3421 gttctagagc ggccgcttca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg
     3481 tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta
     3541 tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt
     3601 cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc
     3661 aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat
     3721 gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc
     3781 tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt
     3841 ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc
     3901 cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc tcttatgaag
     3961 atccctcgac ctggagctcc agcttttgtt ccctttagtg agggttaatt tcgagcttgg
     4021 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     4081 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     4141 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4201 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     4261 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4321 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4381 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4441 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4501 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4561 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4621 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4681 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4741 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4801 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4861 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4921 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4981 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5041 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5101 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5161 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5221 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5281 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5341 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5401 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5461 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5521 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5581 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5641 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5701 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5761 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5821 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5881 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5941 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6001 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6061 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6121 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6181 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     6241 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     6301 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     6361 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     6421 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     6481 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     6541 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     6601 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg
     6661 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     6721 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     6781 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg gtacctagta
     6841 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     6901 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     6961 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     7021 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     7081 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     7141 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag
     7201 ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt
     7261 atttattttt taattatttt gtgcagcgat gggggcgggg ggggggggrc tttccaaaat
     7321 gtcgtaatac ccccgccccg ttgacgcaaa tgggcaagct tgccgggtcg aggtaggcgt
     7381 gtacggtggg aggcctatat aagcaaccgg tataatcaaa cagaccagat tgtctgtttg
     7441 ttaccggtgt ttagtgaacc gggcgcgcct catatcgcct ggagacgcca tccacgctgt
     7501 tttgacctcc atagaag
//
