LOCUS       sequence-364904-        7059 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of a fluorescent sensor for leukotriene B4 in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7059)
  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 7059)
  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-LTB
FEATURES             Location/Qualifiers
     source          1..7059
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      4..459
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(91..110)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     301..322
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     623..642
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        623..641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        706..1009
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1011..1286
                     /label=chicken beta-actin promoter
     intron          1287..2303
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2227..2249
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2311..2330
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     2359..2375
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2360..2376
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3284..3305
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3411..3432)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4291..4307)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4291..4307)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4396..4415)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4461..4516
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4515..4534)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     promoter        complement(4871..4889)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4910..4926)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4910..4926)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4923..4945)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4934..4950
                     /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(4958..4964,4965..4982,4983..4988))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5003..5024
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5141..5158)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5312..5900)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5392..5411)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6071..6862,6863..6931))
                     /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     6694..6713
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6932..7036)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
       61 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
      121 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      181 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
      241 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
      301 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
      361 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
      421 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg gtacctagta
      661 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
      721 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
      781 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
      841 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
      901 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
      961 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag
     1021 ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt
     1081 atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggg sgcgcgccag
     1141 gcggggcggg gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca
     1201 atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg cggcggccct
     1261 ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc
     1321 gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta ctcccacagg
     1381 tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt taatgacggc
     1441 ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc cctttgtgcg
     1501 gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt ggggagcgcc gcgtgcggct
     1561 ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc ggggctttgt gcgctccgca
     1621 gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg gtgcgggggg ggctgcgagg
     1681 ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg agcagggggt gtgggcgcgt
     1741 cggtcgggct gcaacccccc ctgcaccccc ctccccgagt tgctgagcac ggcccggctt
     1801 cgggtgcggg gctccgtacg gggcgtggcg cggggctcgc cgtgccgggc ggggggtggc
     1861 ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc cggggagggc tcgggggagg
     1921 ggcgcggcgg cccccggagc gccggcggct gtcgaggcgc ggcgagccgc agccattgcc
     1981 ttttatggta atcgtgcgag agggcgcagg gacttccttt gtcccaaatc tgtgcggagc
     2041 cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg cggggcgaag cggtgcggcg
     2101 ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc
     2161 tccctctcca gcctcggggc tgtccgcggg gggacggctg ccttcggggg ggacggggca
     2221 gggcggggtt cggcttctgg cgtgtgaccg gcggctctag agcctctgct aaccatgttc
     2281 atgccttctt ctttttccta cagctcctgg gcaacgtgct ggttattgtg ctgtctcatc
     2341 attttggcaa aggattcctc gaggtcgacg gtatcgataa gcttgatatc gaattcacca
     2401 tggcatccac cggaaacatc agcgtttctt tgcagccgcc ctccagatcc atgagcacct
     2461 ctgccctggt cggggtcgcc atcctgtctc tggcatttct gctgggcttt cctgggaact
     2521 tgtttgtggt ttggtcggtg ctctgccggg tgaagaagcg ctcggtgacg tgtctgttgg
     2581 tgctgaacct ggcgatggca gacgccttcg tcctgctgag cgcccctctt ttcctcagat
     2641 acctggctgg aggaaggggc tgggagtttg gcaccgtggc gtgcaagctg gtgcattacc
     2701 tgtcaagtgt gaacatgtac gtgtccattt acctgatctg tctgatgagc atggaccgct
     2761 ggctggctgt ctcgaggcct ttcatgtctc atagaatgag gaccaagcgc tcgctcctgg
     2821 ttctcctgct gggagtttgg gtgttagcgt tcctgctgtc actgccgatg cccttttacc
     2881 gcagtaatgt gaagaaaccg atggggaacg gcaccattct gtactattgc ataccctatc
     2941 actggaggag tgtccctcac caggttttcc agtacctgtt tgagaccatc atgggctgca
     3001 ttgtgccctt ttcgctcatc agtagctgtt actcgtcggt ctgctacaaa atatggcgaa
     3061 atttcaaagg gaaattaaac gtctatatca aggccgacaa gcagaagaac ggcatcaagg
     3121 cgaacttcca catccgccac aacatcgagg acggcggcgt gcagctcgcc taccactacc
     3181 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagcg
     3241 tgcagaccaa actttcgaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     3301 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggc ggtaccggag
     3361 ggatgatggt gagaaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     3421 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggtgag ggcgatgcca
     3481 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     3541 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     3601 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacatccag gagcgcacca
     3661 tcttcttcaa ggacgacggc cactacaaga cccgcgccga ggtgaagttc gagggcgaca
     3721 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     3781 ggcacaagct ggagtacaac tcagtaatct ccaaggcaaa gatacgcaca gtgaaaatga
     3841 tcttgctcat cctctcagct tttgttgttt tctggctgcc ttatcacatc atcaacatca
     3901 ttgaggtggc cggcctcctg cagaacagtg agtcagtgat caaagctgct aagacagcac
     3961 ggcccaatgt tatagccttc gcatacttca gcagtgccat caaccccatc ctttacgtct
     4021 tcgctgggag ctcccacatt cgcctggctg ggtttagttt catgggcaaa cttttcgaag
     4081 caaccaactc tgagagcaga agcacgtcca tcttcagccg tggtcgcaac agctcctctc
     4141 cagatgagat gtctgtcctg cactcggtgt cggttaaact tgggaagcct tttaagagca
     4201 agagcaaggg gaggagtagc agtgtggcgg gcaaagagga caacgagccg gagctgaaga
     4261 cgctggccac tgttgaacag tgtgaataag gatccactag ttctagagcg gccgcttcac
     4321 tcctcaggtg caggctgcct atcagaaggt ggtggctggt gtggccaatg ccctggctca
     4381 caaataccac tgagatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct
     4441 tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa
     4501 ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa
     4561 tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg
     4621 ttggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca
     4681 tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc
     4741 tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg
     4801 actactccca gtcatagctg tccctcttct cttatgaaga tccctcgacc tggagctcca
     4861 gcttttgttc cctttagtga gggttaattt cgagcttggc gtaatcatgg tcatagctgt
     4921 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4981 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     5041 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     5101 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     5161 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5221 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5281 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5341 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5401 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5461 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5521 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5581 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5641 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5701 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     5761 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5821 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     5881 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5941 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6001 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6061 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6121 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6181 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6241 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6301 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6361 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6421 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6481 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6541 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6601 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6661 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6721 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6781 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6841 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6901 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6961 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7021 aaataggggt tccgcgcaca tttccccgaa aagtgccac
//
