LOCUS       sequence-240666-        6482 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the genetically-encoded fluorescent norepinephrine(NE) 
            sensor GRAB_NE1m in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6482)
  AUTHORS   Feng J, Zhang C, Lischinsky JE, Jing M, Zhou J, Wang H, Zhang Y, 
            Dong A, Wu Z, Wu H, Chen W, Zhang P, Zou J, Hires SA, Zhu JJ, Cui G,
            Lin D, Du J, Li Y
  TITLE     A Genetically Encoded Fluorescent Sensor for Rapid and Specific In 
            Vivo Detection of Norepinephrine.
  JOURNAL   Neuron. 2019 May 22;102(4):745-761.e8. doi: 
            10.1016/j.neuron.2019.02.037. Epub 2019 Mar 25.
  PUBMED    30922875
REFERENCE   2  (bases 1 to 6482)
  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"; doi: 
            "10.1016/j.neuron.2019.02.037"; journalName: "Neuron"; date: 
            "2019-05-22- 22"; volume: "102"; issue: "4"; pages: "745-761.e8"
COMMENT     Sequence Label: pAAV-hSyn-GRAB_NE1m
FEATURES             Location/Qualifiers
     source          1..6482
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   74..203
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      654..663
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     660..719
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     protein_bind    723..747
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     1850..1871
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1977..1998)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2238..2257)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(2735..2755)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3153..3164)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3302..3778
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3501..3520)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3818..3958
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3818..3947
                     /label=AAV2 ITR
     rep_origin      4033..4488
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4120..4139)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4330..4351
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4505..4524)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4624..4646
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4684..4702)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4770..4874
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4875..4943,4944..5735)
                     /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(5093..5112)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5906..6482,1..12)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6395..6414
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
       61 tttgctcaca tgtcctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg
      121 tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc
      181 caactccatc actaggggtt cctgcggccg cacgcgtatg aggcggggtg ggggtgccta
      241 cctgacgacc gaccccgacc cactggacaa gcacccaacc cccattcccc aaattgcgca
      301 tcccctatca gagaggggga ggggaaacag gatgcggcga ggcgcgtgcg cactgccagc
      361 ttcagcaccg cggacagtgc cttcgccccc gcctggcggc gcgcgccacc gccgcctcag
      421 cactgaaggc gcgctgacgt cactcgccgg tcccccgcaa actccccttc ccggccacct
      481 tggtcgcgtc cgcgccgccg ccggcccagc cggaccgcac cacgcgaggc gcgagatagg
      541 ggggcacggg cgcgaccatc tgcgctgcgg cgccggcgac tcagcgctgc ctcagtctgc
      601 ggtgggcagc ggaggagtcg tgtcgtgcct gagagcgcag tcgagaaacc ggtgccacca
      661 tggagacaga cacactcctg ctatgggtac tgctgctctg ggttccaggt tccactggtg
      721 acacaagttt gtacaaaaaa gttggcacca ccggtggctc cctgcagccg gacgcgggca
      781 acgcgagctg gaacgggacc gaggcgccgg ggggcggcgc ccgggccacc ccttactccc
      841 tgcaggtgac gctgacgctg gtgtgcctgg ccggcctgct catgctgctc accgtgttcg
      901 gcaacgtgct cgtcatcatc gccgtgttca cgagccgcgc gctcaaggcg ccccaaaacc
      961 tcttcctggt gtctctggcc tcggccgaca tcctggtggc cacgctcgtc atccctttct
     1021 cgctggccaa cgaggtcatg ggctactggt acttcggcaa ggcttggtgc gagatctacc
     1081 tggcgctcga cgtgctcttc tgcacgtcgt ccatcgtgca cctgtgcgcc atcagcctgg
     1141 accgctactg gtccatcaca caggccatcg agtacaacct gaagcgcacg ccgcgccgca
     1201 tcaaggccat catcatcacc gtgtgggtca tctcggccgt catctccttc ccgccgctca
     1261 tctccatcga gaagaagggc ggcggcggcg gcccgcagcc ggccgagccg cgctgcgaga
     1321 tcaacgacca gaagtggtac gtcatctcgt cgtgcatcgg ctccttcttc gctccctgcc
     1381 tcatcatgat cctggtctac gtgcgcatct accagatcgc caagcgtcgc acccgcgtgc
     1441 cacccagccg ccggggtccg gacgccgtcg ccgcgccgcc ggggggcacc gagcgcaggc
     1501 ccaacggtct gggccccgag cgcagcgcgg gcccgggggg cgcagaggcc gaaccgctgc
     1561 ccacccagct caacggcgcc cctggcgagc ccgcgccggc cgggccgcgc gacaccgacg
     1621 cgctggacct ggaggaggga ggaaacgtct atatcaaggc cgacaagcag aagaacggca
     1681 tcaaggcgaa cttccacatc cgccacaaca tcgaggacgg cggcgtgcag ctcgcctacc
     1741 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     1801 tgagcgtgca gtccaaactt tcgaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     1861 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagggcggta
     1921 ccggagggag catggtgaga aagggcgagg agctgttcac cggggtggtg cccatcctgg
     1981 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggtgagggcg
     2041 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     2101 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     2161 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac atccaggagc
     2221 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     2281 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     2341 tcctggggca caagctggag tacaacaccg gagcagcagc acgctggcgc gggcggcaga
     2401 accgcgagaa gcgcttcacg ttcgtgctgg ccgtggtcat cggagtgttc gtggtgtgct
     2461 ggttcccctt cttcttcacc tacacgctca cggccgtcgg gtgctccgtg ccacgcacgc
     2521 tcttcaaatt cttcttctgg ttcggctact gcaacagctc gttgaacccg gtcatctaca
     2581 ccatcttcaa ccacgatttc cgccgcgcct tcaagaagat cctctgtcgg ggggacagga
     2641 agcggatcgt gttgtgagaa ttcgatatca agcttatcga tcatcaacct ctggattaca
     2701 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     2761 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     2821 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     2881 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     2941 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     3001 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     3061 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctatgtt gccacctgga
     3121 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     3181 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     3241 gtcggatctc cctttgggcc gcctccccgc atcgataccg agcgctgctc gagagatcta
     3301 cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc
     3361 agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc
     3421 cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac
     3481 aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg
     3541 gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt
     3601 gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg
     3661 gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc
     3721 ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct
     3781 gattttgtag gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg
     3841 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     3901 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
     3961 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     4021 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     4081 gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt
     4141 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     4201 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     4261 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     4321 tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt
     4381 cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt
     4441 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     4501 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     4561 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     4621 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     4681 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     4741 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4801 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4861 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     4921 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     4981 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5041 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     5101 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5161 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     5221 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5281 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5341 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5401 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     5461 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     5521 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     5581 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5641 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     5701 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     5761 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5821 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5881 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     5941 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6001 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     6061 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     6121 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6181 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6241 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6301 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6361 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6421 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6481 gg
//
