LOCUS       sequence-415628-        5351 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-vector for fluorescent calcium measurements in the green 
            spectrum from noradrenergic neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5351)
  AUTHORS   Wissing C, Eschholz LS, Maheu M, Sauter K, Morellini F, Wiegert JS, 
            Dieter A
  TITLE     A comparison of viral strategies and model systems to target 
            norepinephrine neurons in the locus coeruleus reveals high 
            variability in transgene expression patterns.
  JOURNAL   PLoS Biol. 2025 Jul 7;23(7):e3003228. doi: 
            10.1371/journal.pbio.3003228. eCollection 2025 Jul.
  PUBMED    40623068
REFERENCE   2  (bases 1 to 5351)
  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"; journalName: "PLoS 
            Biol."; date: "2025-07-7"; volume: "23(7):e3003228. doi"; pages: " 
            10.1371/journal.pbio.3003228. eCollection 2025 Jul"
COMMENT     Sequence Label: PRS_8-GCaMP8m
FEATURES             Location/Qualifiers
     source          1..5351
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             419..436
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     718..739
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(845..866)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1106..1125)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    1697..2285
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1750..1770)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2168..2179)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2454..2535
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2491..2510)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2545..2564
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   2614..2754
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2614..2743
                     /label=AAV2 ITR
     rep_origin      2829..3284
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2916..2935)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3126..3147
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3301..3320)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3420..3442
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3480..3498)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3566..3670
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3671..3739,3740..4531)
                     /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(3889..3908)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4702..5290
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5191..5210
                     /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 gcgtttaatt aaaaacgcgt attagcttcc gctagacaaa
      181 tgtgattacc cccgctagac aaatgtgatt acccgcgcta gacaaatgtg attaccccgc
      241 tagacaaatg tgattacccc ccgctagaca aatgtgatta cccccgctag acaaatgtga
      301 ttacccgcgc tagacaaatg tgattacccc gctagacaaa tgtgattacc cccgaccagg
      361 gcataaatgg ccaggtggga ccagagatat cgctagcaag gatccacccg ccaccatgca
      421 tcatcaccat catcacacgc gtcgcaagaa gaccttcaag gaggtggcca ccgccgtgaa
      481 gatcatcggc cgcctgatgg gcctgaagat caacgtctat atcaaggccg acaagcagaa
      541 gaacggcatc aaggcgaact tccacatccg ccacaacatc gaggacggcg gcgtgcagct
      601 cgcctaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
      661 ccactacctg agcgtggagt ccaaactttc gaaagacccc aacgagaagc gcgatcacat
      721 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
      781 gggcggtacc ggagggagca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
      841 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
      901 tgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
      961 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     1021 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacat
     1081 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     1141 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     1201 cggcaacatc ctggggcaca agctggagta caacctgccg gaccaactga ctgaagagca
     1261 gatcgcagaa tacaaagagg ctttctccct atttgacaag gacggggatg ggacaataac
     1321 aaccaaggag ctggggacgg tgatgcggtc tctggggcag aaccccacag aagcagagct
     1381 gcaggacatg atcaatgaag tagatgccga cggtgacggc acaatcgact tccctgagtt
     1441 cctgacaatg atggcaagaa aaatgaaata cagggacacg gaagaagaaa ttagagaagc
     1501 gttcggtgtg tttgataagg atggcaatgg ctacatcagt gcagcagagc ttcgccacgt
     1561 gatgacaaac cttggagaga agttaacaga tgaagaggtt gatgaaatga tcagggaagc
     1621 agacatcgat ggggatggtc aggtaaacta cgaagagttt gtacaaatga tgacagcgaa
     1681 gtaaaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     1741 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     1801 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     1861 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     1921 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     1981 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     2041 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     2101 ctttccttgg ctgctcgcct atgttgccac ctggattctg cgcgggacgt ccttctgcta
     2161 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     2221 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     2281 cccgcatcga taccgagcgc tgctcgagag atctgtgata gcggccatca agctggccgc
     2341 gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     2401 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     2461 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     2521 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcagcttatc
     2581 gataccgcat gcacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca
     2641 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     2701 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc
     2761 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
     2821 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     2881 cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt
     2941 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3001 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     3061 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3121 taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt
     3181 tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca
     3241 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
     3301 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     3361 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     3421 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     3481 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     3541 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3601 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3661 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     3721 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     3781 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     3841 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     3901 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     3961 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     4021 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     4081 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     4141 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     4201 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     4261 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     4321 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     4381 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     4441 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     4501 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     4561 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     4621 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     4681 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     4741 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4801 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     4861 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4921 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4981 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     5041 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     5101 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     5161 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     5221 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     5281 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     5341 tgctcacatg t
//
