LOCUS       sequence-240667-        6482 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the genetically-encoded fluorescent norepinephrine(NE) 
            control sensor GRAB_NEmut 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_NEmut
FEATURES             Location/Qualifiers
     source          1..6482
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   68..197
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      648..657
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     654..713
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     protein_bind    717..741
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     1844..1865
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1971..1992)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2232..2251)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2676..3264
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2729..2749)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3147..3158)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3296..3772
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3495..3514)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3812..3952
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3812..3941
                     /label=AAV2 ITR
     rep_origin      4027..4482
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4114..4133)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4324..4345
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4499..4518)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4618..4640
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4678..4696)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4764..4868
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4869..4937,4938..5729)
                     /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(5087..5106)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5900..6482,1..6)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6389..6408
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
       61 cacatgtcct gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg ggcgtcgggc
      121 gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc
      181 catcactagg ggttcctgcg gccgcacgcg tatgaggcgg ggtgggggtg cctacctgac
      241 gaccgacccc gacccactgg acaagcaccc aacccccatt ccccaaattg cgcatcccct
      301 atcagagagg gggaggggaa acaggatgcg gcgaggcgcg tgcgcactgc cagcttcagc
      361 accgcggaca gtgccttcgc ccccgcctgg cggcgcgcgc caccgccgcc tcagcactga
      421 aggcgcgctg acgtcactcg ccggtccccc gcaaactccc cttcccggcc accttggtcg
      481 cgtccgcgcc gccgccggcc cagccggacc gcaccacgcg aggcgcgaga taggggggca
      541 cgggcgcgac catctgcgct gcggcgccgg cgactcagcg ctgcctcagt ctgcggtggg
      601 cagcggagga gtcgtgtcgt gcctgagagc gcagtcgaga aaccggtgcc accatggaga
      661 cagacacact cctgctatgg gtactgctgc tctgggttcc aggttccact ggtgacacaa
      721 gtttgtacaa aaaagttggc accaccggtg gctccctgca gccggacgcg ggcaacgcga
      781 gctggaacgg gaccgaggcg ccggggggcg gcgcccgggc caccccttac tccctgcagg
      841 tgacgctgac gctggtgtgc ctggccggcc tgctcatgct gctcaccgtg ttcggcaacg
      901 tgctcgtcat catcgccgtg ttcacgagcc gcgcgctcaa ggcgccccaa aacctcttcc
      961 tggtgtctct ggcctcggcc gacatcctgg tggccacgct cgtcatccct ttctcgctgg
     1021 ccaacgaggt catgggctac tggtacttcg gcaaggcttg gtgcgagatc tacctggcgc
     1081 tcgacgtgct cttctgcacg tcgtccatcg tgcacctgtg cgccatcagc ctggaccgct
     1141 actggtccat cacacaggcc atcgagtaca acctgaagcg cacgccgcgc cgcatcaagg
     1201 ccatcatcat caccgtgtgg gtcatctcgg ccgtcatctc cttcccgccg ctcatctcca
     1261 tcgagaagaa gggcggcggc ggcggcccgc agccggccga gccgcgctgc gagatcaacg
     1321 accagaagtg gtacgtcatc tcgtcgtgca tcggcgcctt cttcgctccc tgcctcatca
     1381 tgatcctggt ctacgtgcgc atctaccaga tcgccaagcg tcgcacccgc gtgccaccca
     1441 gccgccgggg tccggacgcc gtcgccgcgc cgccgggggg caccgagcgc aggcccaacg
     1501 gtctgggccc cgagcgcagc gcgggcccgg ggggcgcaga ggccgaaccg ctgcccaccc
     1561 agctcaacgg cgcccctggc gagcccgcgc cggccgggcc gcgcgacacc gacgcgctgg
     1621 acctggagga gggaggaaac gtctatatca aggccgacaa gcagaagaac ggcatcaagg
     1681 cgaacttcca catccgccac aacatcgagg acggcggcgt gcagctcgcc taccactacc
     1741 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagcg
     1801 tgcagtccaa actttcgaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     1861 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggc ggtaccggag
     1921 ggagcatggt gagaaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     1981 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggtgag ggcgatgcca
     2041 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     2101 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     2161 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacatccag gagcgcacca
     2221 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     2281 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     2341 ggcacaagct ggagtacaac accggagcag cagcacgctg gcgcgggcgg cagaaccgcg
     2401 agaagcgctt cacgttcgtg ctggccgtgg tcatcggagt gttcgtggtg tgctggttcc
     2461 ccttcttctt cacctacacg ctcacggccg tcgggtgctc cgtgccacgc acgctcttca
     2521 aattcttctt ctggttcggc tactgcaaca gctcgttgaa cccggtcatc tacaccatct
     2581 tcaaccacga tttccgccgc gccttcaaga agatcctctg tcggggggac aggaagcgga
     2641 tcgtgttgtg agaattcgat atcaagctta tcgataatca acctctggat tacaaaattt
     2701 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     2761 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     2821 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     2881 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     2941 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     3001 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     3061 tgtcggggaa atcatcgtcc tttccttggc tgctcgccta tgttgccacc tggattctgc
     3121 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     3181 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     3241 tctccctttg ggccgcctcc ccgcatcgat accgagcgct gctcgagaga tctacgggtg
     3301 gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc
     3361 caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta
     3421 taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg
     3481 tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac
     3541 tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg
     3601 attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt
     3661 caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc
     3721 tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt
     3781 gtaggtaacc acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact
     3841 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     3901 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg
     3961 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
     4021 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     4081 tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc
     4141 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     4201 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     4261 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     4321 atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg
     4381 atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa
     4441 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
     4501 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     4561 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     4621 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     4681 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     4741 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4801 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4861 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     4921 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     4981 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     5041 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     5101 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     5161 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     5221 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     5281 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     5341 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     5401 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     5461 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     5521 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     5581 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     5641 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     5701 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     5761 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5821 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5881 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5941 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     6001 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     6061 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     6121 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     6181 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     6241 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     6301 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     6361 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     6421 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6481 ta
//
