LOCUS       sequence-364387-        7125 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of a fluorescent sensor for neuropeptide FF in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7125)
  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 7125)
  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-NPFF
FEATURES             Location/Qualifiers
     source          1..7125
                     /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     2663..2684
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2790..2811)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3652..3668)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3652..3668)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3757..3776)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3822..3877
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3876..3895)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     promoter        complement(4232..4250)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4271..4287)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4271..4287)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4284..4306)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4295..4311
                     /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(4319..4325,4326..4343,4344..4349))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4364..4385
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4502..4519)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4673..5261)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4753..4772)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5432..6223,6224..6292))
                     /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     6055..6074
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6293..6397)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6424..6879
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6511..6530)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6721..6742
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7005..7027
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7019..7036
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7020..7036
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7043..7062
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7043..7061
                     /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 ggaggggcgt 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 caccatgagc gagaaatggg actcaaactc ttcagaaagc tggaatcaca
     1741 tctggagtgg caatgataca cagcatcact ggtattcaga tatcaacatt acctatgtga
     1801 actactatct ccaccagccc caagtggcag ctgtcttcat cagctcctac ctcctgatct
     1861 ttgtcttgtg catggtggga aatactgtcg tttgctttat tgtgataagg aatagacaca
     1921 tgcacacagt cactaatttc ttcatcttaa accttgccat aagtgattta ctggttggaa
     1981 tattctgtat gcctatcaca ttgctggaca acatcatagc aggatggcca ttcggaagca
     2041 gcatgtgcaa gatcagtggg ctagtgcaag ggatatcagt tgcggcttcc gtcttcacct
     2101 tggttgcaat agctgtggac agattccgct gtgtggtcta cccctttaag ccaaagctca
     2161 ctgtcaagac agcctttgtc acgattgtga tcatctgggg cctggccatc gccattatga
     2221 ctccatctgc aataatgtta catgtacaag aagaaaaata ctaccgtgtg agactcagct
     2281 cccacaataa aaccagcaca gtctactggt gtcgggagga ctggccaaga cacgaaatga
     2341 ggaggatcta taccacggtg ctatttgcca ccatctatct tgctcctctc tcactcattg
     2401 ttatcatgta tgcaaggatt tgctacaaaa tatggcgaaa tttcaaaggg aaattaaacg
     2461 tctatatcaa ggccgacaag cagaagaacg gcatcaaggc gaacttccac atccgccaca
     2521 acatcgagga cggcggcgtg cagctcgcct accactacca gcagaacacc cccatcggcg
     2581 acggccccgt gctgctgccc gacaaccact acctgagcgt gcagaccaaa ctttcgaaag
     2641 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     2701 ctctcggcat ggacgagctg tacaagggcg gtaccggagg gatgatggtg agaaagggcg
     2761 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     2821 acaagttcag cgtgtccggc gagggtgagg gcgatgccac ctacggcaag ctgaccctga
     2881 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     2941 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     3001 agtccgccat gcccgaaggc tacatccagg agcgcaccat cttcttcaag gacgacggcc
     3061 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     3121 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     3181 cagtaatctc caaggcaaag atacgcacag tgaaaatgct gctgactgtg gccctccttt
     3241 tcatcctttc ctggcttccc ctgtggaccc tgatgatgct ctcagactat actgacctgt
     3301 ctcctaacaa actgcgtatc atcaacatct acatctaccc tttcgcccac tggctcgcct
     3361 tctgcaacag cagtgtcaac cctattattt atggattctt taatgaaaat tttcgcaatg
     3421 gtttccaaga tgctttccag atctgccaaa agaaagccaa gccccaggaa gcctattccc
     3481 tgagagcgaa acgcaacata gtcataaaca catcgggcct gctggtgcag gaaccggtgt
     3541 ctcaaaaccc aggtggggaa aatttgggat gtggaaaaag tgcagacaat cccacacagg
     3601 aatccttgat agaggaaatg ggagaagcta ctaacagtac tgtggcttaa ggatccacta
     3661 gttctagagc ggccgcttca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg
     3721 tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta
     3781 tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt
     3841 cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc
     3901 aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat
     3961 gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc
     4021 tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt
     4081 ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc
     4141 cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc tcttatgaag
     4201 atccctcgac ctggagctcc agcttttgtt ccctttagtg agggttaatt tcgagcttgg
     4261 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     4321 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     4381 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4441 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     4501 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4561 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4621 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4681 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4741 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4801 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4861 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4921 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4981 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5041 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5101 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5161 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5221 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5281 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5341 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5401 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5461 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5521 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5581 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5641 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5701 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5761 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5821 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5881 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5941 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6001 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6061 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     6121 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6181 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6241 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6301 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6361 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6421 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     6481 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     6541 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     6601 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     6661 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     6721 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     6781 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     6841 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg
     6901 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     6961 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     7021 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg gtacctagta
     7081 atcaattacg gggtcattag ttcatagccc atatatggag ttccg
//
