LOCUS       sequence-292569-        7044 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For packaging and expression of Chronos-mT-Sapphire featuring a 
            neuritin (NRN) 3' UTR in neurons via AAV.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7044)
  AUTHORS   Christoffel DJ, Walsh JJ, Hoerbelt P, Heifets BD, Llorach P, Lopez 
            RC, Ramakrishnan C, Deisseroth K, Malenka RC
  TITLE     Selective filtering of excitatory inputs to nucleus accumbens by 
            dopamine and serotonin.
  JOURNAL   Proc Natl Acad Sci U S A. 2021 Jun 15;118(24). pii: 2106648118. doi:
            10.1073/pnas.2106648118.
  PUBMED    34103400
REFERENCE   2  (bases 1 to 7044)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2021-06-15"; volume: "118(24). pii: 
            2106648118. doi"; pages: " 10.1073/pnas.2106648118"
COMMENT     Sequence Label: pAAV-hSyn-Chronos-mT-Sapphire-NRN
FEATURES             Location/Qualifiers
     source          1..7044
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        236..683
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      696..705
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1692..1694,1695..2405)
                     /codon_start=1
                     /product="strongly enhanced green fluorescent protein 
                     (Kremers et al., 2007)"
                     /label=SGFP2
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVMVFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYIMADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSIQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1734..1755)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1995..2014)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2342..2363
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2440..3028
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2493..2513)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2911..2922)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4054..4071)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4060..4267
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4307..4447
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4307..4436
                     /label=AAV2 ITR
     rep_origin      4522..4977
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4609..4628)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4819..4840
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4994..5013)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5113..5135
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5173..5191)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5259..5363
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5364..5432,5433..6224)
                     /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(5582..5601)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6395..6983
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6884..6903
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtttaac attatggcct taggtcactt
      181 catctccatg gggttcttct tctgattttc tagactgcag agggccctgc gtatgagtgc
      241 aagtgggttt taggaccagg atgaggcggg gtgggggtgc ctacctgacg accgaccccg
      301 acccactgga caagcaccca acccccattc cccaaattgc gcatccccta tcagagaggg
      361 ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca ccgcggacag
      421 tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa ggcgcgctga
      481 cgtcactcgc cggtcccccg caaactcccc ttcccggcca ccttggtcgc gtccgcgccg
      541 ccgccggccc agccggaccg caccacgcga ggcgcgagat aggggggcac gggcgcgacc
      601 atctgcgctg cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc agcggaggag
      661 tcgtgtcgtg cctgagagcg cagtcgagag gatccgccac catggaaaca gccgccacaa
      721 tgacccacgc ctttatctca gccgtgccta gcgccgaagc cacaattaga ggcctgctga
      781 gcgccgcagc agtggtgaca ccagcagcag acgctcacgg agaaacctct aacgccacaa
      841 cagccggagc cgatcacggt tgcttccccc acatcaacca cggaaccgag ctgcagcaca
      901 agatcgcagt gggactccag tggttcaccg tgatcgtggc tatcgtgcag ctcatcttct
      961 acggttggca cagcttcaag gccacaaccg gctgggagga ggtctacgtc tgcgtgatcg
     1021 agctcgtcaa gtgcttcatc gagctgttcc acgaggtcga cagcccagcc acagtgtacc
     1081 agaccaacgg aggagccgtg atttggctgc ggtacagcat gtggctcctg acttgccccg
     1141 tgatcctgat ccacctgagc aacctgaccg gactgcacga agagtacagc aagcggacca
     1201 tgaccatcct ggtgaccgac atcggcaaca tcgtgtgggg gatcacagcc gcctttacaa
     1261 agggccccct gaagatcctg ttcttcatga tcggcctgtt ctacggcgtg acttgcttct
     1321 tccagatcgc caaggtgtat atcgagagct accacaccct gcccaaaggc gtctgccgga
     1381 agatttgcaa gatcatggcc tacgtcttct tctgctcttg gctgatgttc cccgtgatgt
     1441 tcatcgccgg acacgaggga ctgggcctga tcacacctta caccagcgga atcggccacc
     1501 tgatcctgga tctgatcagc aagaacactt ggggcttcct gggccaccac ctgagagtga
     1561 agatccacga gcacatcctg atccacggcg acatccggaa gacaaccacc atcaacgtgg
     1621 ccggcgagaa catggagatc gagaccttcg tcgacgagga ggagggagga gtggcggcac
     1681 cggtagtagc agtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     1741 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     1801 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     1861 ggcccaccct cgtgaccacc ttctcctacg gcgtgatggt gttcgcccgc taccccgacc
     1921 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     1981 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     2041 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     2101 tggggcacaa gctggagtac aacttcaaca gccacaacgt ctatatcatg gccgacaagc
     2161 agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcggcgtgc
     2221 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     2281 acaaccacta cctgagcatc cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc
     2341 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctgggcatg gacgagctgt
     2401 acaagtccgg actcagatct cgagctcaag cttatcgata atcaacctct ggattacaaa
     2461 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     2521 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     2581 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     2641 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     2701 tgtcagctcc tttccggaac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     2761 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     2821 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctatgttgc cacctggatt
     2881 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     2941 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     3001 cggatctccc tttgggccgc ctccccgcat cgataccgag cgctgctcga gagatctgcg
     3061 tggggccagc tccccccgcg cgcccaccca cactcactcc atgctcccgg aaatcgagag
     3121 gaagatccat tagttctttg gggacgttgt gattctctgt gatgctgaaa acactcatat
     3181 aggattgtgg gaaatcctga ttctcttttt tatttcgttt gatttcttgt gttttatttg
     3241 ccaaatgtta ccaatcagtg agcaagcaag cacagccaaa atcggacctc agctttagtc
     3301 cgtcttcaca cacaaataag aaaacggcaa acccacccca ttttttaatt ttattattat
     3361 taattttttt tgttggcaaa agaatctcag gaacggccct gggccaccta ctatattaat
     3421 catgctagta acatgaaaaa tgatgggctc ctcctaatag gaaggcgagg agaggagaag
     3481 gccaggggaa tgaattcaag agagatgtcc acggacgaaa catacggtga ataattcacg
     3541 ctcacgtcgt tcttccacag tatcttgttt tgatcatttc cactgcacat ttctcctcaa
     3601 gaaaagcgaa aggacagact gttggctttg tgtttggagg ataggaggga gagagggaag
     3661 gggctgagga aatctctggg gtaagagtaa aggcttccag aagacatgct gctatggtca
     3721 ctgaggggtt agctttatct gctgttgttg atgcatccgt ccaagttcac tgcctttatt
     3781 ttccctcctc cctcttgttt tagctgttac acacacagta atacctgaat atccaacggt
     3841 atagatcaca agggggggat gttaaatgtt aatttaaaat atagctaaaa aaagattttg
     3901 acataaaaga gccttgattt taaaaaaaaa agagagagag atgtaattta aaaagtttat
     3961 tataaattaa attcagcaaa aaaagatttg ctacaaagta tagagaagta taaaataaaa
     4021 gttattgttt gaaaaaaaaa aaaaaaaaat aagcctcgac tgtgccttct agttgccagc
     4081 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     4141 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     4201 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     4261 ctggggactg attttgtagg taaccacgtg cggaccgagc ggccgcagga acccctagtg
     4321 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     4381 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
     4441 ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     4501 atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4561 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4621 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4681 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     4741 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     4801 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     4861 cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     4921 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat
     4981 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     5041 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     5101 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     5161 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     5221 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     5281 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     5341 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     5401 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     5461 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     5521 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     5581 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     5641 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     5701 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5761 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     5821 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5881 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     5941 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     6001 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     6061 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     6121 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     6181 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     6241 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6301 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6361 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6421 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6481 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6541 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6601 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6661 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6721 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6781 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6841 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6901 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6961 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     7021 tttgctggcc ttttgctcac atgt
//
