LOCUS       sequence-357977-        7056 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of a fluorescent sensor for norepinephrine in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7056)
  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 7056)
  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-NE
FEATURES             Location/Qualifiers
     source          1..7056
                     /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     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        709..1012
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1015..1290
                     /label=chicken beta-actin promoter
     intron          1291..2299
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2223..2245
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2355..2371
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2356..2372
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3304..3325
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3431..3452)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(3992..4003)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4395..4414)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4460..4515
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4514..4533)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4868..4888)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4868..4886)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4907..4923)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4907..4923)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4920..4942)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4931..4947
                     /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(4955..4961,4962..4979,4980..4985))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5000..5021
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5138..5155)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5309..5897)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5389..5408)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6068..6859,6860..6928))
                     /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     6691..6710
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6929..7033)
                     /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 gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccta
      661 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      721 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      781 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggacta
      841 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      901 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
      961 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tgggtcgagg
     1021 tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc ccaattttgt
     1081 atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggggg ggggcgcgcg
     1141 ccaggcgggg cggggcgggg cgaggggcgg ggcggggcga ggcggagagg tgcggcggca
     1201 gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg
     1261 ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg cgttgccttc gccccgtgcc
     1321 ccgctccgcg ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag
     1381 gtgagcgggc gggacggccc ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg
     1441 ctcgtttctt ttctgtggct gcgtgaaagc cttaaagggc tccgggaggg ccctttgtgc
     1501 gggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg
     1561 cccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc
     1621 gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg gtgcgggggg gctgcgaggg
     1681 gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga gcagggggtg tgggcgcggc
     1741 ggtcgggctg taaccccccc ctgcaccccc ctccccgagt tgctgagcac ggcccggctt
     1801 cgggtgcggg gctccgtgcg gggcgtggcg cggggctcgc cgtgccgggc ggggggtggc
     1861 ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc cggggagggc tcgggggagg
     1921 ggcgcggcgg ccccggagcg ccggcggctg tcgaggcgcg gcgagccgca gccattgcct
     1981 tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg tcccaaatct ggcggagccg
     2041 aaatctggga ggcgccgccg caccccctct agcgggcgcg ggcgaagcgg tgcggcgccg
     2101 gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc
     2161 atctccagcc tcggggctgc cgcaggggga cggctgcctt cgggggggac ggggcagggc
     2221 ggggttcggc ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc atgttcatgc
     2281 cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt gttgtgctgt ctcatcattt
     2341 tggcaaagga ttcctcgagg tcgacggtat cgataagctt gatatcgaat tcaccatggg
     2401 gccacacggg aacgacagcg acttcttgct ggcacccaac ggaagccgag cgccagacca
     2461 cgacgtcact caggaacggg acgaagcgtg ggttgtgggc atggccatcc tcatgtcggt
     2521 tatcgtcctg gccatcgtgt ttggcaacgt gctggtcatc acagccattg ccaagttcga
     2581 gcgactacaa accgtcacca actacttcat aatctccttg gcgtgtgctg atctagtcat
     2641 gggcctagcg gtggtgccgt ttggggccag tcacatcctt atgaaaatgt ggaattttgg
     2701 caacttctgg tgcgagttct ggacttccat tgatgtgttg tgcgtcacag ccagcatcga
     2761 gaccctgtgc gtgattgcag tggatcgcta tgttgctatc acatcgccct tcaagtacca
     2821 gagcctgctg accaagaata aggcccgagt ggtcatcctg atggtatgga ttgtatctgg
     2881 ccttacctcc tttttgccta tccagatgca ctggtaccgt gccacccaca agaaagctat
     2941 cgattgttac accgaggaga cttgctgtga cttcttcacg aaccaggcct acgccatcgc
     3001 gtcctcgatt gtgtctttct acgtgcccct ggtggtgatg gtctttgtct attcccgggt
     3061 ctgctacaaa atatggcgaa atttcaaagg gaaattaaac gtctatatca aggccgacaa
     3121 gcagaagaac ggcatcaagg cgaacttcca catccgccac aacatcgagg acggcggcgt
     3181 gcagctcgcc taccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     3241 cgacaaccac tacctgagcg tgcagaccaa actttcgaaa gaccccaacg agaagcgcga
     3301 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     3361 gtacaagggc ggtaccggag ggatgatggt gagaaagggc gaggagctgt tcaccggggt
     3421 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     3481 cgagggtgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     3541 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     3601 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     3661 ctacatccag gagcgcacca tcttcttcaa ggacgacggc cactacaaga cccgcgccga
     3721 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     3781 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tcagtaatct ccaaggcaaa
     3841 gatacgcaca gtgaaaatgt taggcatcat catgggcaca ttcaccctct gctggctgcc
     3901 cttcttcatt gtcaatatcg tgcacgttat cagggacaac ctcatcccta aggaagttta
     3961 catcctcctt aactggttgg gctacgtcaa ctctgccttc aatcctctta tctactgtcg
     4021 gagtccagat ttcaggattg cctttcaaga gcttctgtgc cttcgcaggt cttcttcgaa
     4081 aacctatggg aacggctact ctagcaatag caacggcaga acggactaca caggggagcc
     4141 aaacacttgt cagctggggc aggagagaga acaggaactg ctgtgtgagg atcccccagg
     4201 catggaaggc tttgtgaact gtcaaggtac tgtgcctagc cttagcgttg actcccaagg
     4261 aaggaactgt agtacaaatg actcgccact gtaaactagt tctagagcgg ccgcttcact
     4321 cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac
     4381 aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt
     4441 gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat
     4501 tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat
     4561 gagtatttgg tttagagttt ggcaacatat gccatatgct ggctgccatg aacaaaggtg
     4621 gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag
     4681 aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt
     4741 aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact
     4801 actcccagtc atagctgtcc ctcttctctt atgaagatcc ctcgacctgg agctccagct
     4861 tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc
     4921 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4981 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     5041 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5101 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5161 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5221 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5281 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5341 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5401 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5461 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5521 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5581 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5641 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5701 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     5761 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5821 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5881 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5941 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6001 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6061 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6121 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6181 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6241 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6301 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6361 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6421 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6481 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6541 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6601 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6661 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6721 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6781 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6841 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6901 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6961 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7021 taggggttcc gcgcacattt ccccgaaaag tgccac
//
