LOCUS       sequence-369597-        6388 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses green norepinephrine indicator nLightG in neuronal cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6388)
  AUTHORS   Kagiampaki Z, Rohner V, Kiss C, Curreli S, Dieter A, Wilhelm M, 
            Harada M, Duss SN, Dernic J, Bhat MA, Zhou X, Ravotto L, Ziebarth T,
            Wasielewski LM, Sonmez L, Benke D, Weber B, Bohacek J, Reiner A, 
            Wiegert JS, Fellin T, Patriarchi T
  TITLE     Sensitive multicolor indicators for monitoring norepinephrine in 
            vivo.
  JOURNAL   Nat Methods. 2023 Sep;20(9):1426-1436. doi: 
            10.1038/s41592-023-01959-z.
  PUBMED    37474807
REFERENCE   2  (bases 1 to 6388)
  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"; journalName: 
            "Nat Methods"; date: "2023-09"; volume: "20"; issue: "9"; pages: 
            "1426-1436"
COMMENT     Sequence Label: pAAV_hSyn1_nLightG
FEATURES             Location/Qualifiers
     source          1..6388
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        164..611
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     intron          635..767
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     750..769
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1737..1758
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1864..1885)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2125..2144)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2767..3355
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2820..2840)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3238..3249)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3373..3597
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3640..3780
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3640..3769
                     /label=AAV2 ITR
     rep_origin      3855..4310
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3942..3961)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4152..4173
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4327..4346)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4446..4468
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4506..4524)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4592..4696
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4697..4765,4766..5557)
                     /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(4915..4934)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5728..6316
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6217..6236
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(6378..6388,1..119)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcgtcgg gcgacctttg gtcgcccggc
       61 ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctg
      121 cggccgcacg cgtgtgtcta gactgcagag ggccctgcgt atgagtgcaa gtgggtttta
      181 ggaccaggat gaggcggggt gggggtgcct acctgacgac cgaccccgac ccactggaca
      241 agcacccaac ccccattccc caaattgcgc atcccctatc agagaggggg aggggaaaca
      301 ggatgcggcg aggcgcgtgc gcactgccag cttcagcacc gcggacagtg ccttcgcccc
      361 cgcctggcgg cgcgcgccac cgccgcctca gcactgaagg cgcgctgacg tcactcgccg
      421 gtcccccgca aactcccctt cccggccacc ttggtcgcgt ccgcgccgcc gccggcccag
      481 ccggaccgca ccacgcgagg cgcgagatag gggggcacgg gcgcgaccat ctgcgctgcg
      541 gcgccggcga ctcagcgctg cctcagtctg cggtgggcag cggaggagtc gtgtcgtgcc
      601 tgagagcgca gtcgagatta attaactcga gcaggtaagt atcaaggtta caagacaggt
      661 ttaaggagac caatagaaac tgggcttgtc gagacagaga agactcttgc gtttctgata
      721 ggcacctatt ggtcttactg acatccactt tgcctttctc tccacaggtg ctagcgttta
      781 aacggtaccg gatcctccgc caccatgaag acgatcatcg ccctgagcta catcttctgc
      841 ctggtgttcg ccgactacaa ggacgatgat gacgcagtct ttctcagtgg gaacgctagt
      901 gactcttcaa attgtaccca cccaccacct cctgttaata tcagcaaggc tattttgctg
      961 ggtgtcatct tgggaggcct gattctgttc ggagtgcttg gtaacattct cgtaattctc
     1021 tcagtagcgt gtcaccgcca tctccacagc gttacccact actatattgt gaacctcgct
     1081 gtagcagacc tgcttctcac gagtacggta ctgccatttt ccgctatctt cgaaattttg
     1141 gggtattggg cctttggacg ggttttttgt aacatctggg ccgctgtgga cgtattgtgt
     1201 tgtacggcat caatcatggg cctgtgcctt atctcaatag atcgatactg ggctatctcc
     1261 agccctgccc ggtatgagag aaagatgacc cccaaggcag ccttcatcgc tctcctttgt
     1321 gtctgggctc tttccttggt tatatccatt gggcccttgt tcggctggcg ccagcccgcc
     1381 ccggaggacg aaacaatatg tcagatcaac gaagagcctg gatacgtcct tttcagcgcc
     1441 ctcggatctt tttatgtccc gttgacaatc atcctggtca tgtattgcag agtttatagg
     1501 attgctcaga aacagctgag ctcactcatt aacgtctata tcaaggccga caagcagaag
     1561 aacggcatca aggcgaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc
     1621 gcctaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     1681 cactacctga gcgtgcagtc caaactttcg aaagacccca acgagaagcg cgatcacatg
     1741 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     1801 ggcggtaccg gagggagcat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     1861 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggt
     1921 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     1981 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     2041 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacatc
     2101 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     2161 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     2221 ggcaacatcc tggggcacaa gctggagtac aacaatcatg accaactgaa aagagaaact
     2281 aaagtcgcta agacgctcgg gatagttgtt ggttgtttcg tactctgctg gttgcccttt
     2341 ttcttggtga tgccgatagg tagctttttt cccgacttcc gaccgagtga gacagtattc
     2401 aaaatagctt tttggcttgg ttatttgaac tcctgtatta atcccattat atatccctgt
     2461 tcatctcaag agtttaaaaa ggcttttcaa aacgtgttgc gcatacaatg cctgcgcaga
     2521 aagcagtctt ctaagcacgc actcggttat acgctccacc ccccatccca tgctctcgag
     2581 ggacagtata gaaacctcgt gcgaatacct gtgggtagtg gcgaaacatt ttgcaaaatc
     2641 agtaaaacag acggggtatg tgaatggaaa tttttttcat ccttgccacg cggatctgcc
     2701 agaataacag ttcctaagga ccagtccgct tgcacaactg cccgggactt ttaaaagctt
     2761 atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     2821 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     2881 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     2941 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     3001 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     3061 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     3121 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     3181 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     3241 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     3301 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgctctcg
     3361 agatacgtac gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     3421 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     3481 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     3541 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     3601 ctgaggcgga aagatctcac gtgcggaccg agcggccgca ggaaccccta gtgatggagt
     3661 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     3721 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
     3781 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     3841 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3901 gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc
     3961 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4021 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     4081 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4141 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta
     4201 ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat
     4261 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     4321 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     4381 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     4441 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     4501 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     4561 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4621 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4681 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4741 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4801 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     4861 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     4921 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     4981 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     5041 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     5101 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     5161 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     5221 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     5281 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     5341 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     5401 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     5461 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     5521 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     5581 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5641 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5701 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5761 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5821 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     5881 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5941 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6001 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6061 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6121 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6181 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6241 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6301 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6361 gccttttgct cacatgtcct gcaggcag
//
