LOCUS       sequence-364795-        7062 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 7062)
  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 7062)
  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..7062
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..581
                     /label=chicken beta-actin promoter
     intron          582..1598
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1522..1544
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1606..1625
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     1654..1670
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1655..1671
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2603..2624
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2730..2751)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(3291..3302)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3694..3713)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3759..3814
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3813..3832)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     promoter        complement(4169..4187)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4208..4224)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4208..4224)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4221..4243)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4232..4248
                     /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(4256..4262,4263..4280,4281..4286))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4301..4322
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4439..4456)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4610..5198)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4690..4709)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5369..6160,6161..6229))
                     /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     5992..6011
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6230..6334)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6361..6816
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6448..6467)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6658..6679
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6942..6964
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6956..6973
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6957..6973
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6980..6999
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6980..6998
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 aacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtcgag gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc
      361 cccaattttg tatttattta ttttttaatt attttgtgca gcgatggggg cggggggggg
      421 gggggsgcgc gccaggcggg gcggggcggg gcgaggggcg gggcggggcg aggcggagag
      481 gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc
      541 ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc
      601 ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc tgactgaccg
      661 cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt aattagcgct
      721 tggtttaatg acggcttgtt tcttttctgt ggctgcgtga aagccttgag gggctccggg
      781 agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg cgtgtgtgtg tgcgtgggga
      841 gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg ctgcgggcgc ggcgcggggc
      901 tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg gggcggtgcc ccgcggtgcg
      961 gggggggctg cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg gggtgagcag
     1021 ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca cccccctccc cgagttgctg
     1081 agcacggccc ggcttcgggt gcggggctcc gtacggggcg tggcgcgggg ctcgccgtgc
     1141 cgggcggggg gtggcggcag gtgggggtgc cgggcggggc ggggccgcct cgggccgggg
     1201 agggctcggg ggaggggcgc ggcggccccc ggagcgccgg cggctgtcga ggcgcggcga
     1261 gccgcagcca ttgcctttta tggtaatcgt gcgagagggc gcagggactt cctttgtccc
     1321 aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac cccctctagc gggcgcgggg
     1381 cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt gcgtcgccgc
     1441 gccgccgtcc ccttctccct ctccagcctc ggggctgtcc gcggggggac ggctgccttc
     1501 gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggc tctagagcct
     1561 ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac gtgctggtta
     1621 ttgtgctgtc tcatcatttt ggcaaaggat tcctcgaggt cgacggtatc gataagcttg
     1681 atatcgaatt caccatgggg ccacacggga acgacagcga cttcttgctg gcacccaacg
     1741 gaagccgagc gccagaccac gacgtcactc aggaacggga cgaagcgtgg gttgtgggca
     1801 tggccatcct catgtcggtt atcgtcctgg ccatcgtgtt tgacaacgtg ctggtcatca
     1861 cagccattgc caagttcgag cgactacaaa ccgtcaccaa ctacttcata atctccttgg
     1921 cgtgtgctga tctagtcatg ggcctagcgg tggtgccgtt tggggccagt cacatcctta
     1981 tgaaaatgtg gaattttggc aacttctggt gcgagttctg gacttccatt gatgtgttgt
     2041 gcgtcacagc cagcatcgag accctgtgcg tgattgcagt ggatcgctat gttgctatca
     2101 catcgccctt caagtaccag agcctgctga ccaagaataa ggcccgagtg gtcatcctga
     2161 tggtatggat tgtatctggc cttacctcct ttttgcctat ccagatgcac tggtaccgtg
     2221 ccacccacaa gaaagctatc gattgttaca ccgaggagac ttgctgtgac ttcttcacga
     2281 accaggccta cgccatcgcg tcctcgattg tgtctttcta cgtgcccctg gtggtgatgg
     2341 tctttgtcta ttcccgggtc tgctacaaaa tatggcgaaa tttcaaaggg aaattaaacg
     2401 tctatatcaa ggccgacaag cagaagaacg gcatcaaggc gaacttccac atccgccaca
     2461 acatcgagga cggcggcgtg cagctcgcct accactacca gcagaacacc cccatcggcg
     2521 acggccccgt gctgctgccc gacaaccact acctgagcgt gcagaccaaa ctttcgaaag
     2581 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     2641 ctctcggcat ggacgagctg tacaagggcg gtaccggagg gatgatggtg agaaagggcg
     2701 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     2761 acaagttcag cgtgtccggc gagggtgagg gcgatgccac ctacggcaag ctgaccctga
     2821 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     2881 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     2941 agtccgccat gcccgaaggc tacatccagg agcgcaccat cttcttcaag gacgacggcc
     3001 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     3061 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     3121 cagtaatctc caaggcaaag atacgcacag tgaaaatgtt aggcatcatc atgggcacat
     3181 tcaccctctg ctggctgccc ttcttcattg tcaatatcgt gcacgttatc agggacaacc
     3241 tcatccctaa ggaagtttac attctcctta actggttggg ctacgtcaac tctgccttca
     3301 atcctcttat ctactgtcgg agtccagatt tcaggattgc ctttcaagag cttctgtgcc
     3361 ttcgcaggtc ttcttcgaaa acctatggga acggctactc tagcaatagc aacggcagaa
     3421 cggactacac aggggagcca aacacttgtc agctggggca ggagagagaa caggaactgc
     3481 tgtgtgagga tcccccaggc atggaaggct ttgtgaactg tcaaggtact gtgcctagcc
     3541 ttagcgttga ctcccaagga aggaactgta gtacaaatga ctcgccactg taaactagtt
     3601 ctagagcggc cgcttcactc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt
     3661 ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca aaaattatgg
     3721 ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat
     3781 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
     3841 tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct
     3901 ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc
     3961 tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg
     4021 ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag
     4081 atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct tatgaagatc
     4141 cctcgacctg gagctccagc ttttgttccc tttagtgagg gttaatttcg agcttggcgt
     4201 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4261 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4321 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4381 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4441 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4501 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4561 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4621 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4681 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4741 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4801 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4861 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4921 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4981 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5041 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5101 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5161 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5221 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5281 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5341 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5401 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5461 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5521 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5581 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5641 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5701 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5761 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5821 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5881 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5941 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6001 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6061 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6121 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6181 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6241 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6301 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     6361 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     6421 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     6481 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     6541 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     6601 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     6661 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     6721 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     6781 acgcgaattt taacaaaata ttaacgctta caatttccat tcgccattca ggctgcgcaa
     6841 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     6901 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     6961 aacgacggcc agtgaattgt aatacgactc actatagggc gaattgggta cctagtaatc
     7021 aattacgggg tcattagttc atagcccata tatggagttc cg
//
