LOCUS       sequence-379303-        4837 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-vector for fluorescent labeling of noradrenergic neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4837)
  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 4837)
  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-eGFP
FEATURES             Location/Qualifiers
     source          1..4837
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   3..132
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      415..424
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(421..423,424..426,427..1140)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(421..423,424..426,427..1137)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(466..487)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(727..746)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1074..1095
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1185..1773
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1238..1258)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1656..1667)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1942..2023
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1979..1998)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2033..2052
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   2102..2242
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2102..2231
                     /label=AAV2 ITR
     rep_origin      2317..2772
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2404..2423)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2614..2635
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2789..2808)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2908..2930
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2968..2986)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3054..3158
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3159..3227,3228..4019)
                     /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(3377..3396)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4190..4778
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4679..4698
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtcctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcgt cgggcgacct
       61 ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca
      121 ctaggggttc ctgcggccgc acgcgtttaa ttaaaaacgc gtattagctt ccgctagaca
      181 aatgtgatta cccccgctag acaaatgtga ttacccgcgc tagacaaatg tgattacccc
      241 gctagacaaa tgtgattacc ccccgctaga caaatgtgat tacccccgct agacaaatgt
      301 gattacccgc gctagacaaa tgtgattacc ccgctagaca aatgtgatta cccccgacca
      361 gggcataaat ggccaggtgg gaccagagat atcgctagca aggatccact agtagccacc
      421 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
      481 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      541 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
      601 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
      661 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      721 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
      781 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
      841 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
      901 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
      961 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     1021 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     1081 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     1141 aggcgcgcca cccctgcagg gaattcgata tcaagcttat cgataatcaa cctctggatt
     1201 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     1261 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     1321 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     1381 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     1441 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     1501 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     1561 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctat gttgccacct
     1621 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     1681 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     1741 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgagcgctg ctcgagagat
     1801 ctgtgatagc ggccatcaag ctggccgcga ctctagatca taatcagcca taccacattt
     1861 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     1921 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     1981 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     2041 tccaaactca tcaatgtatc agcttatcga taccgcatgc acgtgcggac cgagcggccg
     2101 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     2161 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     2221 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc
     2281 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
     2341 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc
     2401 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     2461 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     2521 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     2581 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     2641 cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct
     2701 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     2761 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     2821 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     2881 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     2941 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     3001 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     3061 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3121 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     3181 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     3241 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     3301 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     3361 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     3421 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     3481 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     3541 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     3601 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     3661 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     3721 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     3781 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     3841 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     3901 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     3961 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     4021 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     4081 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     4141 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     4201 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4261 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     4321 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     4381 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     4441 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     4501 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     4561 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     4621 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     4681 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     4741 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     4801 tacggttcct ggccttttgc tggccttttg ctcacat
//
