LOCUS       sequence-372397-        7191 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of a fluorescent sensor for dopamine in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7191)
  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 7191)
  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-DA
FEATURES             Location/Qualifiers
     source          1..7191
                     /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"
     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     3350..3371
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3477..3498)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4423..4439)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4423..4439)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4528..4547)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4593..4648
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4647..4666)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     promoter        complement(5003..5021)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5042..5058)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5042..5058)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5055..5077)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5066..5082
                     /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(5090..5096,5097..5114,5115..5120))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5135..5156
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5273..5290)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5444..6032)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5524..5543)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6203..6994,6995..7063))
                     /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     6826..6845
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7064..7168)
                     /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 sgcgcgccrg
     1141 gsggggsggg gsgggggggg gggggggggg gggsgrggsg grgrggkgsg 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 cggggrgggc 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 tggagaatcc agccaagtat ctctcttcgg tgcagggggt tgattctgcg ctggcacccc
     2461 tcggagagat tatgtggaac agcactggag cggaggcaac aagcgatgcg gggaaggata
     2521 tggtggtgcg cacggtgacg ggctgtctgt tgtccctgct catcctctgg accctgctgg
     2581 ggaacatcct ggtgtgctcc gcggtgctgc gctttcggca cttgcggacc aaagtcacta
     2641 acatttttat cgtgtccctg gctctgtcgg atttatttgt cgccgtcctg gtgatgccat
     2701 ggaaagcagt ggcagaggtg gcagggtact ggccgtttgg cactttttgt aatgtctggg
     2761 ttgcttttga tatcatgtgt tccactgcct ccatccttaa cctctgcatc attagtgtgg
     2821 atagatactg ggcaatatcg agccctttcc gttacgagag gaaaatgacc cagcgagttg
     2881 cctttgttat gataagcatc acttggacgt tgtcagtgct catttcattc ataccggtcc
     2941 agctaaactg gcacaaagcc agcgctgagg atagggcgca caactccaca agttggcagt
     3001 ttgaggaaaa ctgcgactcc agcctcagta gagagtatgc tatatcctcc tccctgataa
     3061 gtttctacat tcctgtagca attatgattg taacatacac gagaatctgc tacaaaatat
     3121 ggcgaaattt caaagggaaa ttaaacgtct atatcaaggc cgacaagcag aagaacggca
     3181 tcaaggcgaa cttccacatc cgccacaaca tcgaggacgg cggcgtgcag ctcgcctacc
     3241 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     3301 tgagcgtgca gaccaaactt tcgaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     3361 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagggcggta
     3421 ccggagggat gatggtgaga aagggcgagg agctgttcac cggggtggtg cccatcctgg
     3481 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggtgagggcg
     3541 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     3601 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     3661 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac atccaggagc
     3721 gcaccatctt cttcaaggac gacggccact acaagacccg cgccgaggtg aagttcgagg
     3781 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     3841 tcctggggca caagctggag tacaactcag taatctccaa ggcaaagata cgcacagtga
     3901 aaatgttgtc ggtgattatg ggcgtctttg tttgttgctg gttacctttc ttcgttctga
     3961 actgcatggt tcctttctgc gagaggcgcg ccacagaccg ggacgcgggt ctgccgtgcg
     4021 ttagcgagac caccttcgac gtgttcgtgt ggttcggctg gaccaactcc tccctgaacc
     4081 ccattattta cgccttcaac gccgagttcc ggaaggcctt cgccagtctg ctgggctgcc
     4141 gtaatttctg ctccaacacg cccgtggaaa cggtcaacat cagcaacgag cttgtctcct
     4201 acaaccagga caccctgatc cacaaggaga tcgcgaacgc ttacgtcaac atgatcccca
     4261 acgtggttga atgtatagaa cacgaggaga cgtttgacag gatctcacag ttttcccaca
     4321 acatagacaa cgccaccgac tccgtctgcg acttggagga ctgcgaggca gacatcagtc
     4381 ttgacaggat gtcccccttc acacccaatg gattacactg aggatccact agttctagag
     4441 cggccgcttc actcctcagg tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa
     4501 tgccctggct cacaaatacc actgagatct ttttccctct gccaaaaatt atggggacat
     4561 catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt tcattgcaat
     4621 agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt
     4681 aaaacatcag aatgagtatt tggtttagag tttggcaaca tatgcccata tgctggctgc
     4741 catgaacaaa ggttggctat aaagaggtca tcagtatatg aaacagcccc ctgctgtcca
     4801 ttccttattc catagaaaag ccttgacttg aggttagatt ttttttatat tttgttttgt
     4861 gttatttttt tctttaacat ccctaaaatt ttccttacat gttttactag ccagattttt
     4921 cctcctctcc tgactactcc cagtcatagc tgtccctctt ctcttatgaa gatccctcga
     4981 cctggagctc cagcttttgt tccctttagt gagggttaat ttcgagcttg gcgtaatcat
     5041 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     5101 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     5161 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     5221 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     5281 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     5341 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5401 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5461 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5521 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5581 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5641 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5701 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5761 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     5821 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5881 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5941 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     6001 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6061 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6121 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6181 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6241 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6301 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6361 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6421 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6481 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6541 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6601 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6661 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6721 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6781 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6841 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     6901 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6961 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     7021 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     7081 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     7141 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca c
//
