LOCUS       sequence-364921-        6939 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of a fluorescent sensor for Prolactin-releasing 
            peptide factor in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6939)
  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 6939)
  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-PRLH
FEATURES             Location/Qualifiers
     source          1..6939
                     /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     2633..2654
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2760..2781)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3466..3482)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3466..3482)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3571..3590)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3636..3691
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3690..3709)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     promoter        complement(4046..4064)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4085..4101)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4085..4101)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4098..4120)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4109..4125
                     /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(4133..4139,4140..4157,4158..4163))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4178..4199
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4316..4333)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4487..5075)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4567..4586)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5246..6037,6038..6106))
                     /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     5869..5888
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6107..6211)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6238..6693
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6325..6344)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6535..6556
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6819..6841
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6833..6850
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6834..6850
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6857..6876
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6857..6875
                     /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 caccatggag gcctctggtt ggccgggtgg agaggattct ccagacagtc
     1741 gagtatatga ggtgacggtg gtccaaaact ccacggctcc tcacggtcac ccgttcgcag
     1801 atgtggcctt gctgcagagc ttcaagcccc ttatcatccc ctgttacgtg ctggtggtcc
     1861 tggtgggcgt tttcggcaac tacctgctga tctacgtcat ctgccgcacc aggaagatgc
     1921 acaacgtcac caacttcttc attgggaatc tggcattctc cgacatgctg atgtgtgtga
     1981 cctgtgtccc gttcaccctg gcctacgcct tcagtgccca tggctggacg ttcggccgct
     2041 tcatgtgtta tctggtgttc ctgatccagc cagtcactgt atacgtgtct gttttcacgc
     2101 tcaccgccat cgctgtggac aggtactacg ccacagtgca ccctctgaag aagcgcatct
     2161 cgatggcggc ctgtgcgtat gtgctgtccg gcatctggct gctgtcctgt gtgctggtgg
     2221 ctccggcggt ggcccacacc taccatgtgg agttcagaga tgagggtttg accatctgcg
     2281 aggagttctg gcttggtcag gagacccagc ggctggtgta cgcttacagc acgctattac
     2341 tgacctacat cctcccttta tccgctgtct gcgtctcgta tttgtgcatc tgctacaaaa
     2401 tatggcgaaa tttcaaaggg aaattaaacg tctatatcaa ggccgacaag cagaagaacg
     2461 gcatcaaggc gaacttccac atccgccaca acatcgagga cggcggcgtg cagctcgcct
     2521 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     2581 acctgagcgt gcagaccaaa ctttcgaaag accccaacga gaagcgcgat cacatggtcc
     2641 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagggcg
     2701 gtaccggagg gatgatggtg agaaagggcg aggagctgtt caccggggtg gtgcccatcc
     2761 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggtgagg
     2821 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     2881 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     2941 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacatccagg
     3001 agcgcaccat cttcttcaag gacgacggcc actacaagac ccgcgccgag gtgaagttcg
     3061 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     3121 acatcctggg gcacaagctg gagtacaact cagtaatctc caaggcaaag atacgcacag
     3181 tgaaaatggt gtctctggtg gtggcggcgt tcgctgtttg ctggctgccc atccatgttt
     3241 ttaatgttct gcgcgacatc gatatccgcc tgatcaacaa gcggcacttt ctgctgatcc
     3301 agctgttgtg tcatctgtgc gctatgagct cgtcctgctg caacccgttt ctgtacgcat
     3361 ggcttcatga ccgctttcgt gcagagctgc gaaagatgtt cacctgccac cggcgcattg
     3421 ggatacctgc aaaccactgc gccacggcca gcgttgtact atgaggatcc actagttcta
     3481 gagcggccgc ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc
     3541 caatgccctg gctcacaaat accactgaga tctttttccc tctgccaaaa attatgggga
     3601 catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc
     3661 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca
     3721 tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc
     3781 tgccatgaac aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt
     3841 ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt
     3901 tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac tagccagatt
     3961 tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat gaagatccct
     4021 cgacctggag ctccagcttt tgttcccttt agtgagggtt aatttcgagc ttggcgtaat
     4081 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     4141 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     4201 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     4261 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     4321 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     4381 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4441 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4501 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4561 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4621 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4681 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4741 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4801 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4861 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4921 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4981 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5041 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     5101 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5161 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5221 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5281 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5341 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5401 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5461 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5521 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5581 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5641 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5701 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5761 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5821 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5881 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5941 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6001 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6061 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6121 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6181 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     6241 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     6301 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     6361 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     6421 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     6481 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     6541 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     6601 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     6661 cgaattttaa caaaatatta acgcttacaa tttccattcg ccattcaggc tgcgcaactg
     6721 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     6781 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     6841 gacggccagt gaattgtaat acgactcact atagggcgaa ttgggtacct agtaatcaat
     6901 tacggggtca ttagttcata gcccatatat ggagttccg
//
