LOCUS       sequence-412174-        6476 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses the genetically-encoded fluorescent norepinephrine (NE) 
            sensor GRAB_NE2h in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6476)
  AUTHORS   Feng J, Dong H, Lischinsky J, Zhou J, Deng F, Zhuang C, Miao X, Wang
            H, Xie H, Cui G, Lin D, Li Y
  TITLE     Monitoring norepinephrine release in vivo using next-generation 
            GRABNE sensors
  JOURNAL   bioRxiv 2023.06.22.546075
REFERENCE   2  (bases 1 to 6476)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2023.06.22.546075"
COMMENT     Sequence Label: pAAV-hsyn-NE2h
FEATURES             Location/Qualifiers
     source          1..6476
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      581..590
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     587..649
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTGD"
     protein_bind    650..674
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     1771..1792
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1898..1919)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2159..2178)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2603..3191
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2656..2676)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3074..3085)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3223..3699
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3422..3441)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3739..3879
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3739..3868
                     /label=AAV2 ITR
     rep_origin      3954..4409
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4041..4060)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4251..4272
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4426..4445)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4545..4567
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4605..4623)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4691..4795
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4796..4864,4865..5656)
                     /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(5014..5033)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5827..6415
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6316..6335
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtatgagg cggggtgggg gtgcctacct gacgaccgac
      181 cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag
      241 agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg
      301 acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg
      361 ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc
      421 gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc
      481 gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt gggcagcgga
      541 ggagtcgtgt cgtgcctgag agcgcagtcg agaaaccggt gccaccatgg agacagacac
      601 actcctgcta tgggtactgc tgctctgggt tccaggttcc actggtgaca caagtttgta
      661 caaaaaagtt ggcaccggct ccctgcagcc ggacgcgggc aacgcgagct ggaacgggac
      721 cgaggcgccg gggggcggcg cccgggccac cccttactcc ctgcaggtga cgctgacgct
      781 ggtgtgcctg gccggcctgc tcatgctgct caccgtgttc ggcaacgtgc tcgtcatcat
      841 cgccgtgttc acgagccgcg cgctcaaggc gccccaaaac ctcttcctgg tgtctctggc
      901 ctcggccgac atcctggtgg ccacgctcgt catccctttc tcgctggcca acgaggtcat
      961 gggctactgg tacttcggca aggcttggtg cgagatctac ctggcgctcg acgtgctctt
     1021 ctgcacgtcg tccatcgtgc acctgtgcgc catcagcctg gaccgctact ggtccatcac
     1081 acaggccaaa gagtacaacc tgaagcgcac gccgcgccgc atcaaggcca tcatcatcac
     1141 cgtgtgggtc atctcggccg tcatctcctt cccgccgctc atctccatcg agaagaaggg
     1201 cggcggcggc ggcccgcagc cggccgagcc gcgctgcgag atcaacgacc agaagtggta
     1261 cgtcatctcg tcgtgcatcg gctccttctt cgctccctgc ctcatcatga tcctggtcta
     1321 cgtgcgcatc taccagatcg ccaagcgtcg cacccgcgtg ccacccagcc gccggggtcc
     1381 ggacgccgtc gccgcgccgc cggggggcac cgagcgcagg cccaacggtc tgggccccga
     1441 gcgcagcgcg ggcccggggg gcgcagaggc cgaaccgctg cccacccagc tcaacggcgc
     1501 ccctggcgag cccgcgccgg ccgggccgcg cgacaccgac gcgctggacc tggaggaggg
     1561 aggaaacgtc tatatcaagg ccgacaagca gaagaacggc atcaaggcga acttccacat
     1621 ccgccacaac atcgaggacg gcggcgtgca gctcgcctac cactaccagc agaacacccc
     1681 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcgtgc agtccaaact
     1741 ttcgaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     1801 cgggatcact ctcggcatgg acgagctgta caagggcggt accggaggga gcatggtgag
     1861 aaagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     1921 aaacggccac aagttcagcg tgtccggcga gggtgagggc gatgccaccg agggcaagct
     1981 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     2041 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     2101 cttcttcaag tccgccatgc ccgaaggcta catccaggag cgcaccatct tcttcaagga
     2161 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     2221 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     2281 gtacaacacc ggagcagcag cacgctggcg cgggcggcag aaccgcgaga agcgcttcat
     2341 tttcgtgctg gccgtggtca tcggagtgtt cgtggtgtgc tggttcccct tcttcttcac
     2401 ctacacgctc acggccgtcg ggtgctccgt gccacgcacg ctcttcaaat tcttcttctg
     2461 gttcggctac tgcaacagct cgttgaaccc ggtcatctac accatcttca accacgattt
     2521 ccgccgcgcc ttcaagaaga tcctctgtcg gggggacagg aagcggatcg tgttgtgaga
     2581 attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     2641 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     2701 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     2761 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     2821 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     2881 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     2941 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     3001 atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt
     3061 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     3121 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     3181 cgcctccccg catcgatacc gagcgctgct cgagagatct acgggtggca tccctgtgac
     3241 ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc
     3301 ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg
     3361 tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg
     3421 gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc
     3481 tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc
     3541 atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc
     3601 aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg
     3661 ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg
     3721 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     3781 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     3841 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc
     3901 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     3961 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4021 tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4081 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4141 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     4201 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4261 gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat
     4321 tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4381 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     4441 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     4501 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     4561 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     4621 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     4681 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4741 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4801 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4861 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4921 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4981 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     5041 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5101 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5161 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5221 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5281 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5341 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5401 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5461 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5521 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5581 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5641 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     5701 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5761 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5821 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5881 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5941 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     6001 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6061 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6121 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6181 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6241 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6301 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6361 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6421 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
