LOCUS       sequence-364869-        7023 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of a fluorescent sensor for vasopressin in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7023)
  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 7023)
  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-AVP
FEATURES             Location/Qualifiers
     source          1..7023
                     /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     2717..2738
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2844..2865)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3655..3674)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3720..3775
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3774..3793)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     promoter        complement(4130..4148)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4169..4185)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4169..4185)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4182..4204)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4193..4209
                     /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(4217..4223,4224..4241,4242..4247))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4262..4283
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4400..4417)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4571..5159)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4651..4670)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5330..6121,6122..6190))
                     /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     5953..5972
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6191..6295)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6322..6777
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6409..6428)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6619..6640
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6903..6925
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6917..6934
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6918..6934
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6941..6960
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6941..6959
                     /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 caccatggaa agcatcaatg tggagagaga ctgggacggt ttaagccttt
     1741 catcactgaa cagcgcagac agaaacaact cttcctcctt tttcgtgtct gacctgaact
     1801 ccttcaacgg ttctcacagt ggaggatccc tgtttgggat ctttgcagaa aatggctcca
     1861 acactactcc tcacgccctg ccccagccca ggataaggga cctgaccctg gctcgagctg
     1921 agattgcagt tctcggcata gtgctggccc tcaccactct tgggaacagc tttgtcttgt
     1981 gggtgctttt gaggagaagg aaacacaatg cacctatgca cctgttcatg gtgaacctgt
     2041 gtgtggccga cctggtggtg gcactgtttc aggttcttcc ccagttgata tgggacatta
     2101 cggaacgttt tcaggggccg gactttctct gccgatccgt caagtattta caaattgtgg
     2161 ggatgttcgc ctcgtcttat atgatagtcg ccatgacggt ggaccggcac cgtgccatct
     2221 gttgcccctt gcaggcctac cgtgtcgggg caatgtcccg ctggaacacc ccagtcatgg
     2281 tggcctgggt gctggcacta gtgctcagca tacctcaggt gttcatcttc tctcgatctg
     2341 aagtggctcc tggagagttc gagtgctggg gacacttcac agagccgtgg gggctcaaag
     2401 cctacgtcac ctggatgacc atggctgtgt tcctgctgcc tgccctcatc attaccatct
     2461 gccagataag gatttgctac aaaatatggc gaaatttcaa agggaaatta aacgtctata
     2521 tcaaggccga caagcagaag aacggcatca aggcgaactt ccacatccgc cacaacatcg
     2581 aggacggcgg cgtgcagctc gcctaccact accagcagaa cacccccatc ggcgacggcc
     2641 ccgtgctgct gcccgacaac cactacctga gcgtgcagac caaactttcg aaagacccca
     2701 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     2761 gcatggacga gctgtacaag ggcggtaccg gagggatgat ggtgagaaag ggcgaggagc
     2821 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     2881 tcagcgtgtc cggcgagggt gagggcgatg ccacctacgg caagctgacc ctgaagttca
     2941 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     3001 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     3061 ccatgcccga aggctacatc caggagcgca ccatcttctt caaggacgac ggccactaca
     3121 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     3181 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactcagtaa
     3241 tctccaaggc aaagatacgc acagtgaaaa tgacgcttgt catcgtgttg gtgtacacaa
     3301 tctgctggtc accgttcttc atagtccagc tgtgggcagc ctgggatccc aaccctccag
     3361 accaaggagt ggccttcacc atcctgatgc tgctggccag tctgaactcg tgcactaacc
     3421 cttggatcta cacagctttt tccagcagtg tgtccagaga gcttcagaac cttctacact
     3481 gccggtcacg acttggccga aggggttccc tgcacgatga ctctactacc acacacacct
     3541 tcaccaccaa ggacaaccta tactgagcgg ccgcttcact cctcaggtgc aggctgccta
     3601 tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact gagatctttt
     3661 tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga cttctggcta
     3721 ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg
     3781 aaggacatat gggagggcaa atcatttaaa acatcagaat gagtatttgg tttagagttt
     3841 ggcaacatat gcccatatgc tggctgccat gaacaaaggt tggctataaa gaggtcatca
     3901 gtatatgaaa cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg
     3961 ttagattttt tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc
     4021 cttacatgtt ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt
     4081 ccctcttctc ttatgaagat ccctcgacct ggagctccag cttttgttcc ctttagtgag
     4141 ggttaatttc gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     4201 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     4261 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4321 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4381 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4441 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4501 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4561 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4621 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4681 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4741 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4801 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4861 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4921 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4981 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5041 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5101 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5161 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5221 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5281 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5341 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5401 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5461 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5521 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5581 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5641 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5701 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5761 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5821 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     5881 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5941 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6001 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6061 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6121 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6181 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6241 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6301 ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     6361 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     6421 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     6481 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     6541 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     6601 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     6661 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     6721 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcca
     6781 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     6841 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     6901 ttcccagtca cgacgttgta aaacgacggc cagtgaattg taatacgact cactataggg
     6961 cgaattgggt acctagtaat caattacggg gtcattagtt catagcccat atatggagtt
     7021 ccg
//
