LOCUS       sequence-364394-        6975 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of a fluorescent sensor for neuropeptide Y in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6975)
  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 6975)
  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-NPY
FEATURES             Location/Qualifiers
     source          1..6975
                     /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     2648..2669
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2775..2796)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3607..3626)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3672..3727
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3726..3745)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     promoter        complement(4082..4100)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4121..4137)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4121..4137)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4134..4156)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4145..4161
                     /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(4169..4175,4176..4193,4194..4199))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4214..4235
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4352..4369)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4523..5111)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4603..4622)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5282..6073,6074..6142))
                     /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     5905..5924
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6143..6247)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6274..6729
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6361..6380)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6571..6592
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6855..6877
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6869..6886
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6870..6886
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6893..6912
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6893..6911
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata 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 caccatggat ccagaagatc aactaaacat gagtcaactg gaagagtctt
     1741 accttgatac tctgaactgc tgctccactg cactctcatt gaatgacgag agtcttctga
     1801 agctggatga cagtacaaaa ctggttggag ttcaggttat tctcatcctg gcttacagca
     1861 ccatcatcct gtttggagtc accggaaact ccttggtggt ttacgtcgtc tacaagttta
     1921 ggaatcttcg aactgtgaca aacttcttta tcataaacct ggctgtggct gacctgctgg
     1981 ttaacactct gtgtttgccc ttcaccctta tctacacact ctatgatgag tggatgtttg
     2041 ggcaggtttt gtgctacctg ctgccctttg ctcaaggcat gaccgtgcag gtctccacca
     2101 tcactttaaa catcattgcc ctggaccggc acaggagcat cgtgcaccac atggagacta
     2161 agatgtccaa agacatgtgt gccatggtta ttgtcttctc ctgggtgatc agcgctttgc
     2221 tggcaagtcc tcttgccatt ttcagggagt acgggtcatt tgacttttca ccagaggagt
     2281 ccattcaggt gtgcacagag aagtggccag gaagcagcat gaacggacgg ctctacagca
     2341 tgtcaatgct ggtggtgcaa gtcgccatac ctttggccat caactctgtt gcatacattc
     2401 gcatctgcta caaaatatgg cgaaatttca aagggaaatt aaacgtctat atcaaggccg
     2461 acaagcagaa gaacggcatc aaggcgaact tccacatccg ccacaacatc gaggacggcg
     2521 gcgtgcagct cgcctaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     2581 tgcccgacaa ccactacctg agcgtgcaga ccaaactttc gaaagacccc aacgagaagc
     2641 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     2701 agctgtacaa gggcggtacc ggagggatga tggtgagaaa gggcgaggag ctgttcaccg
     2761 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     2821 ccggcgaggg tgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     2881 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     2941 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     3001 aaggctacat ccaggagcgc accatcttct tcaaggacga cggccactac aagacccgcg
     3061 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     3121 tcaaggagga cggcaacatc ctggggcaca agctggagta caactcagta atctccaagg
     3181 caaagatacg cacagtgaaa atgatgctga ccatggtggt ggtttttgcc gtctgctggt
     3241 tgcctttcca cgcctttcag ctggcaattg acattgataa caccgttctg cacatgaagg
     3301 actttaagct ggtcttcact gttttccaca tcttggcgat gtgctcaacg tgcgtcaatc
     3361 ccatcctgta tggatggatg aacaacaact acaggacggc ttttttgacc gtgtgcaaat
     3421 gttgtggctc cctcactgtg aactccacaa gcagagaggc acaaaaagac agggtttgta
     3481 ctgatgataa atcgacaaac gtctaaacta gttctagagc ggccgcttca ctcctcaggt
     3541 gcaggctgcc tatcagaagg tggtggctgg tgtggccaat gccctggctc acaaatacca
     3601 ctgagatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct
     3661 gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg
     3721 tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt
     3781 ggtttagagt ttggcaacat atgcccatat gctggctgcc atgaacaaag gttggctata
     3841 aagaggtcat cagtatatga aacagccccc tgctgtccat tccttattcc atagaaaagc
     3901 cttgacttga ggttagattt tttttatatt ttgttttgtg ttattttttt ctttaacatc
     3961 cctaaaattt tccttacatg ttttactagc cagatttttc ctcctctcct gactactccc
     4021 agtcatagct gtccctcttc tcttatgaag atccctcgac ctggagctcc agcttttgtt
     4081 ccctttagtg agggttaatt tcgagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     4141 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     4201 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     4261 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     4321 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     4381 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4441 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4501 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4561 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4621 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4681 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4741 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4801 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4861 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4921 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     4981 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5041 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5101 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5161 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5221 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5281 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5341 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5401 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5461 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5521 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5581 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5641 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5701 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5761 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5821 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5881 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5941 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6001 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6061 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6121 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6181 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6241 ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg
     6301 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     6361 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     6421 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     6481 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     6541 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     6601 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     6661 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     6721 ttacaatttc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     6781 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     6841 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat tgtaatacga
     6901 ctcactatag ggcgaattgg gtacctagta atcaattacg gggtcattag ttcatagccc
     6961 atatatggag ttccg
//
