LOCUS       sequence-379794-        7283 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For production of AAV for a fluorescent sensor for oxytocin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7283)
  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 7283)
  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: pAAV.EF1a.FLEX.MTRIA-OT
FEATURES             Location/Qualifiers
     source          1..7283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   19..148
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        171..382
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          383..1321
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1278..1298
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1330..1363)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(1414..1447)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     1456..1472
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1456..1472
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     2229..2250
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2356..2377)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3330..3346)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3331..3347)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     protein_bind    3349..3382
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    3433..3466
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3481..4069
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3534..3554)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3952..3963)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4293..4312)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4564..4704
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4564..4693
                     /label=AAV2 ITR
     rep_origin      4779..5234
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4866..4885)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5076..5097
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5251..5270)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5370..5392
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5430..5448)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5516..5620
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5621..5689,5690..6481)
                     /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     complement(5839..5858)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6652..7240
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7141..7160
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggccttttgc tcacatgtcc tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc
       61 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
      121 gtggccaact ccatcactag gggttcctgc ggccgcaatt gggtaccagt gggcagagcg
      181 cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa ccggtgccta
      241 gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc
      301 cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa
      361 cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc ctggcctctt
      421 tacgggttat ggcccttgcg tgccttgaat tacttccacc tggctgcagt acgtgattct
      481 tgatcccgag cttcgggttg gaagtgggtg ggagagttcg aggccttgcg cttaaggagc
      541 cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc tggggccgcc gcgtgcgaat
      601 ctggtggcac cttcgcgcct gtctcgctgc tttcgataag tctctagcca tttaaaattt
      661 ttgatgacct gctgcgacgc tttttttctg gcaagatagt cttgtaaatg cgggccaaga
      721 tctgcacact ggtatttcgg tttttggggc cgcgggcggc gacggggccc gtgcgtccca
      781 gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg gacgggggta
      841 gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta tcgccccgcc
      901 ctgggcggca aggctggccc ggtcggcacc agttgcgtga gcggaaagat ggccgcttcc
      961 cggccctgct gcagggagct caaaatggag gacgcggcgc tcgggagagc gggcgggtga
     1021 gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat gtgactccac
     1081 ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg agcttttgga gtacgtcgtc
     1141 tttaggttgg ggggaggggt tttatgcgat ggagtttccc cacactgagt gggtggagac
     1201 tgaagttagg ccagcttggc acttgatgta attctccttg gaatttgccc tttttgagtt
     1261 tggatcttgg ttcattctca agcctcagac agtggttcaa agtttttttc ttccatttca
     1321 ggtgtcgaga taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga
     1381 ttgtagctgc tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg
     1441 aagttatgcg gccgctctag aactagtgga tcctcatgtg gtggaagtct gagtaatgct
     1501 tctgctgctg cttgtgctct taatggcgaa gtttgaggag ttgcttttgt gactagaatt
     1561 gaaatctcgc tcgcagcggc actcgggtga cttgaggtag cgagtggagc agcacaggag
     1621 gttctgccga aggtcatgga agaggtggcc ggcaaagcac atgtagatcc aggggttgca
     1681 gcagctgttc aggctcgcca gcagcatgga gatgatgaag ggcatggcct cacggggagc
     1741 cgcttgatcc caggcagacc acatctggac agagaaaaac ggggtccagc agacgatgta
     1801 tgccacgact atcacaaaag tcattttcac tgtgcgtatc tttgccttgg agattactga
     1861 gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag
     1921 ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtg
     1981 gccgtcgtcc ttgaagaaga tggtgcgctc ctggatgtag ccttcgggca tggcggactt
     2041 gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg aagcactgca cgccgtaggt
     2101 cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatgaactt
     2161 cagggtcagc ttgccgtagg tggcatcgcc ctcaccctcg ccggacacgc tgaacttgtg
     2221 gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc
     2281 gccctttctc accatcatcc ctccggtacc gcccttgtac agctcgtcca tgccgagagt
     2341 gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct cgttggggtc
     2401 tttcgaaagt ttggtctgca cgctcaggta gtggttgtcg ggcagcagca cggggccgtc
     2461 gccgatgggg gtgttctgct ggtagtggta ggcgagctgc acgccgccgt cctcgatgtt
     2521 gtggcggatg tggaagttcg ccttgatgcc gttcttctgc ttgtcggcct tgatatagac
     2581 gtttaatttc cctttgaaat ttcgccatat tttgtagcag atcagcccgt aacaaatgct
     2641 caggatggcc actgggatga tgtagatggt gagactgatc catgtgatgt aagctttggc
     2701 cccccagggc ttcacaaagt cgccccagca gtcgtacact cctgacccgg ccgagccaac
     2761 ctctctgagg gagaagatga acatctgcgg gatgctgaag atcaaactca gcaaccatga
     2821 caatatgaca taaattcgat cttttctccg atgcaaggag tgcagaggct gacagatggc
     2881 caaacatctg tcgatggaca ttaaaactaa catgtacgtg gaggcgaaca ttccaaccac
     2941 ctgcaggtat ttcaccaacc tgcacaaaat gtcaggtcca taaaaccgga atgtgatgtc
     3001 ccagataagt tgggggagaa cttgaaatat tgccacaact agatcggcga tgctcaggtg
     3061 cttcatgaag taatacatgc gggactggct gtgtttggtg gtgtggatgg ccagcaggac
     3121 gcacaggttc cccgccagcg ccagaaacag caccagcgcc agcaccgtca cctccacctt
     3181 cgccacctcc tcgttccgct tcagggggtt cgtctgattc agagcgccgg tgccgttacc
     3241 gaggcttgag ttcctccacg atccgttgaa ctgccagatc tcactttcat tggaaatgat
     3301 ctccatggtg aattcgatat caagcttatc gataccgtcg acctcgagat aacttcgtat
     3361 aaagtatcct atacgaagtt atttgcctta acccagaaat tatcactgtt attctttaga
     3421 atggtgcaaa gaataacttc gtataatgta tgctatacga agttatggct agttctagat
     3481 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     3541 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     3601 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     3661 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     3721 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     3781 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     3841 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc
     3901 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     3961 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     4021 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgct agctttctcg
     4081 ctagcagatc tacgggtggc atccctgtga cccctcccca gtgcctctcc tggccctgga
     4141 agttgccact ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc
     4201 tgactaggtg tccttctata atattatggg gtggaggggg gtggtatgga gcaaggggca
     4261 agttgggaag acaacctgta gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt
     4321 ggcacaatct tggctcactg caatctccgc ctcctgggtt caagcgattc tcctgcctca
     4381 gcctcccgag ttgttgggat tccaggcatg catgaccagg ctcagctaat ttttgttttt
     4441 ttggtagaga cggggtttca ccatattggc caggctggtc tccaactcct aatctcaggt
     4501 gatctaccca ccttggcctc ccaaattgct gggattacag gcgtgaacca ctgctgcggc
     4561 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     4621 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     4681 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt
     4741 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     4801 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc
     4861 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4921 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4981 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     5041 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     5101 aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt
     5161 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     5221 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     5281 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     5341 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     5401 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     5461 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     5521 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     5581 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     5641 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5701 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     5761 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     5821 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     5881 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     5941 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     6001 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     6061 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     6121 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     6181 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     6241 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     6301 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     6361 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     6421 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     6481 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     6541 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6601 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6661 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6721 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6781 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6841 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6901 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     6961 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     7021 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     7081 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     7141 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     7201 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     7261 tttacggttc ctggcctttt gct
//
