LOCUS       sequence-240668-        6482 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the genetically-encoded fluorescent norepinephrine(NE) 
            sensor GRAB_NE1h 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_NE1h
FEATURES             Location/Qualifiers
     source          1..6482
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   46..175
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      626..635
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     632..691
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     protein_bind    695..719
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     1822..1843
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1949..1970)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2210..2229)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2654..3242
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2707..2727)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3125..3136)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3274..3750
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3473..3492)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3790..3930
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3790..3919
                     /label=AAV2 ITR
     rep_origin      4005..4460
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4092..4111)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4302..4323
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4477..4496)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4596..4618
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4656..4674)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4742..4846
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4847..4915,4916..5707)
                     /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(5065..5084)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5878..6466
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6367..6386
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctttttacg gttcctggcc ttttgctggc cttttgctca catgtcctgc aggcagctgc
       61 gcgctcgctc gctcactgag gccgcccggg cgtcgggcga cctttggtcg cccggcctca
      121 gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg ttcctgcggc
      181 cgcacgcgta tgaggcgggg tgggggtgcc tacctgacga ccgaccccga cccactggac
      241 aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg gaggggaaac
      301 aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt gccttcgccc
      361 ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac gtcactcgcc
      421 ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc cgccggccca
      481 gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca tctgcgctgc
      541 ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt cgtgtcgtgc
      601 ctgagagcgc agtcgagaaa ccggtgccac catggagaca gacacactcc tgctatgggt
      661 actgctgctc tgggttccag gttccactgg tgacacaagt ttgtacaaaa aagttggcac
      721 caccggtggc tccctgcagc cggacgcggg caacgcgagc tggaacggga ccgaggcgcc
      781 ggggggcggc gcccgggcca ccccttactc cctgcaggtg acgctgacgc tggtgtgcct
      841 ggccggcctg ctcatgctgc tcaccgtgtt cggcaacgtg ctcgtcatca tcgccgtgtt
      901 cacgagccgc gcgctcaagg cgccccaaaa cctcttcctg gtgtctctgg cctcggccga
      961 catcctggtg gccacgctcg tcatcccttt ctcgctggcc aacgaggtca tgggctactg
     1021 gtacttcggc aaggcttggt gcgagatcta cctggcgctc gacgtgctct tctgcacgtc
     1081 gtccatcgtg cacctgtgcg ccatcagcct ggaccgctac tggtccatca cacaggccat
     1141 cgagtacaac ctgaagcgca cgccgcgccg catcaaggcc atcatcatca ccgtgtgggt
     1201 catctcggcc gtcatctcct tcccgccgct catctccatc gagaagaagg gcggcggcgg
     1261 cggcccgcag ccggccgagc cgcgctgcga gatcaacgac cagaagtggt acgtcatctc
     1321 gtcgtgcatc ggctccttct tcgctccctg cctcatcatg atcctggtct acgtgcgcat
     1381 ctaccagatc gccaagcgtc gcacccgcgt gccacccagc cgccggggtc cggacgccgt
     1441 cgccgcgccg ccggggggca ccgagcgcag gcccaacggt ctgggccccg agcgcagcgc
     1501 gggcccgggg ggcgcagagg ccgaaccgct gcccacccag ctcaacggcg cccctggcga
     1561 gcccgcgccg gccgggccgc gcgacaccga cgcgctggac ctggaggagg gaggaaacgt
     1621 ctatatcaag gccgacaagc agaagaacgg catcaaggcg aacttccaca tccgccacaa
     1681 catcgaggac ggcggcgtgc agctcgccta ccactaccag cagaacaccc ccatcggcga
     1741 cggccccgtg ctgctgcccg acaaccacta cctgagcgtg cagtccaaac tttcgaaaga
     1801 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     1861 tctcggcatg gacgagctgt acaagggcgg taccggaggg agcatggtga gaaagggcga
     1921 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     1981 caagttcagc gtgtccggcg agggtgaggg cgatgccacc tacggcaagc tgaccctgaa
     2041 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     2101 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     2161 gtccgccatg cccgaaggct acatccagga gcgcaccatc ttcttcaagg acgacggcaa
     2221 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     2281 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacac
     2341 cggagcagca gcacgctggc gcgggcggca gaaccgcgag aagcgcttca agttcgtgct
     2401 ggccgtggtc atcggagtgt tcgtggtgtg ctggttcccc ttcttcttca cctacacgct
     2461 cacggccgtc gggtgctccg tgccacgcac gctcttcaaa ttcttcttct ggttcggcta
     2521 ctgcaacagc tcgttgaacc cggtcatcta caccatcttc aaccacgatt tccgccgcgc
     2581 cttcaagaag atcctctgtc ggggggacag gaagcggatc gtgttgtgag aattcgatat
     2641 caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     2701 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     2761 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     2821 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     2881 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     2941 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     3001 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     3061 tccttggctg ctcgcctatg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     3121 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     3181 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     3241 gcatcgatac cgagcgctgc tcgagagatc tacgggtggc atccctgtga cccctcccca
     3301 gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa
     3361 ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg
     3421 gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg
     3481 gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt
     3541 caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg
     3601 ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc
     3661 tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag
     3721 gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg
     3781 agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     3841 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     3901 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc
     3961 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg
     4021 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4081 cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4141 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4201 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4261 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4321 tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     4381 ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     4441 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata
     4501 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     4561 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     4621 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     4681 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     4741 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     4801 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     4861 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     4921 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     4981 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     5041 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     5101 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     5161 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     5221 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     5281 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     5341 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     5401 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     5461 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     5521 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     5581 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     5641 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     5701 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     5761 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     5821 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     5881 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     5941 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6001 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     6061 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6121 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6181 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6241 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6301 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6361 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6421 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6481 gg
//
