LOCUS       sequence-412172-        6482 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the genetically-encoded fluorescent norepinephrine (NE) 
            sensor GRAB_NE2m in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6482)
  AUTHORS   Feng J, Dong H, Lischinsky JE, Zhou J, Deng F, Zhuang C, Miao X, 
            Wang H, Li G, Cai R, Xie H, Cui G, Lin D, Li Y
  TITLE     Monitoring norepinephrine release in vivo using next-generation 
            GRAB(NE) sensors.
  JOURNAL   Neuron. 2024 Jun 19;112(12):1930-1942.e6. doi: 
            10.1016/j.neuron.2024.03.001. Epub 2024 Mar 27.
  PUBMED    38547869
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.2024.03.001"; journalName: "Neuron"; date: 
            "2024-06-19- 19"; volume: "112"; issue: "12"; pages: "1930-1942.e6"
COMMENT     Sequence Label: pAAV-hsyn-NE2m
FEATURES             Location/Qualifiers
     source          1..6482
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   15..144
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      595..604
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     601..660
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     protein_bind    664..688
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     1791..1812
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1918..1939)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2179..2198)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2623..3211
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2676..2696)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3094..3105)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3243..3719
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3442..3461)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3759..3899
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3759..3888
                     /label=AAV2 ITR
     rep_origin      3974..4429
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4061..4080)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4271..4292
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4446..4465)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4565..4587
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4625..4643)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4711..4815
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4816..4884,4885..5676)
                     /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(5034..5053)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5847..6435
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6336..6355
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttttgctcac atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc
       61 gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg
      121 ccaactccat cactaggggt tcctgcggcc gcacgcgtat gaggcggggt gggggtgcct
      181 acctgacgac cgaccccgac ccactggaca agcacccaac ccccattccc caaattgcgc
      241 atcccctatc agagaggggg aggggaaaca ggatgcggcg aggcgcgtgc gcactgccag
      301 cttcagcacc gcggacagtg ccttcgcccc cgcctggcgg cgcgcgccac cgccgcctca
      361 gcactgaagg cgcgctgacg tcactcgccg gtcccccgca aactcccctt cccggccacc
      421 ttggtcgcgt ccgcgccgcc gccggcccag ccggaccgca ccacgcgagg cgcgagatag
      481 gggggcacgg gcgcgaccat ctgcgctgcg gcgccggcga ctcagcgctg cctcagtctg
      541 cggtgggcag cggaggagtc gtgtcgtgcc tgagagcgca gtcgagaaac cggtgccacc
      601 atggagacag acacactcct gctatgggta ctgctgctct gggttccagg ttccactggt
      661 gacacaagtt tgtacaaaaa agttggcacc accggtggct ccctgcagcc ggacgcgggc
      721 aacgcgagct ggaacgggac cgaggcgccg gggggcggcg cccgggccac cccttactcc
      781 ctgcaggtga cgctgacgct ggtgtgcctg gccggcctgc tcatgctgct caccgtgttc
      841 ggcaacgtgc tcgtcatcat cgccgtgttc acgagccgcg cgctcaaggc gccccaaaac
      901 ctcttcctgg tgtctctggc ctcggccgac atcctggtgg ccacgctcgt catccctttc
      961 tcgctggcca acgaggtcat gggctactgg tacttcggca aggcttggtg cgagatctac
     1021 ctggcgctcg acgtgctctt ctgcacgtcg tccatcgtgc acctgtgcgc catcagcctg
     1081 gaccgctact ggtccatcac acaggccaaa gagtacaacc tgaagcgcac gccgcgccgc
     1141 atcaaggcca tcatcatcac cgtgtgggtc atctcggccg tcatctcctt cccgccgctc
     1201 atctccatcg agaagaaggg cggcggcggc ggcccgcagc cggccgagcc gcgctgcgag
     1261 atcaacgacc agaagtggta cgtcatctcg tcgtgcatcg gctccttctt cgctccctgc
     1321 ctcatcatga tcctggtcta cgtgcgcatc taccagatcg ccaagcgtcg cacccgcgtg
     1381 ccacccagcc gccggggtcc ggacgccgtc gccgcgccgc cggggggcac cgagcgcagg
     1441 cccaacggtc tgggccccga gcgcagcgcg ggcccggggg gcgcagaggc cgaaccgctg
     1501 cccacccagc tcaacggcgc ccctggcgag cccgcgccgg ccgggccgcg cgacaccgac
     1561 gcgctggacc tggaggaggg aggaaacgtc tatatcaagg ccgacaagca gaagaacggc
     1621 atcaaggcga acttccacat ccgccacaac atcgaggacg gcggcgtgca gctcgcctac
     1681 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     1741 ctgagcgtgc agtccaaact ttcgaaagac cccaacgaga agcgcgatca catggtcctg
     1801 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagggcggt
     1861 accggaggga gcatggtgag aaagggcgag gagctgttca ccggggtggt gcccatcctg
     1921 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggtgagggc
     1981 gatgccaccg agggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     2041 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     2101 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta catccaggag
     2161 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     2221 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     2281 atcctggggc acaagctgga gtacaacacc ggagcagcag cacgctggcg cgggcggcag
     2341 aaccgcgaga agcgcttctt cttcgtgctg gccgtggtca tcggagtgtt cgtggtgtgc
     2401 tggttcccct tcttcttcac ctacacgctc acggccgtcg ggtgctccgt gccacgcacg
     2461 ctcttcaaat tcttcttctg gttcggctac tgcaacagct cgttgaaccc ggtcatctac
     2521 accatcttca accacgattt ccgccgcgcc ttcaagaaga tcctctgtcg gggggacagg
     2581 aagcggatcg tgttgtgaga attcgatatc aagcttatcg ataatcaacc tctggattac
     2641 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     2701 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     2761 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     2821 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     2881 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     2941 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     3001 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctatgt tgccacctgg
     3061 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     3121 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     3181 agtcggatct ccctttgggc cgcctccccg catcgatacc gagcgctgct cgagagatct
     3241 acgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc
     3301 cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt
     3361 ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga
     3421 caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt
     3481 ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt
     3541 tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac
     3601 ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg atctacccac
     3661 cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc
     3721 tgattttgta ggtaaccacg tgcggaccga gcggccgcag gaacccctag tgatggagtt
     3781 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     3841 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg
     3901 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca
     3961 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     4021 cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct
     4081 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4141 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     4201 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4261 ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat
     4321 tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt
     4381 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact
     4441 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     4501 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     4561 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     4621 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     4681 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4741 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4801 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     4861 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     4921 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     4981 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     5041 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     5101 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     5161 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5221 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5281 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5341 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     5401 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     5461 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     5521 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     5581 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     5641 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     5701 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5761 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5821 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5881 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5941 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     6001 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     6061 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6121 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6181 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6241 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6301 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6361 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6421 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6481 cc
//
