LOCUS       sequence-346694-        5793 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector for Flpo recombinase dependent jGCaMP7s.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5793)
  AUTHORS   Jin L, Sullivan HA, Zhu M, Lavin TK, Matsuyama M, Fu X, Lea NE, Xu 
            R, Hou Y, Rutigliani L, Pruner M, Babcock KR, Ip JPK, Hu M, Daigle 
            TL, Zeng H, Sur M, Feng G, Wickersham IR
  TITLE     Long-term labeling and imaging of synaptically connected neuronal 
            networks in vivo using double-deletion-mutant rabies viruses.
  JOURNAL   Nat Neurosci. 2024 Jan 11. doi: 10.1038/s41593-023-01545-8.
  PUBMED    38212587
REFERENCE   2  (bases 1 to 5793)
  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: "Nat 
            Neurosci. 2024 Jan 11. doi: 10.1038/s41593-023-01545-8."
COMMENT     Sequence Label: pAAV-syn-F14F15S-jGCaMP7s
FEATURES             Location/Qualifiers
     source          1..5793
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        164..611
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             complement(join(744..1187,1188..1193,1194..1628,1629..1646,
                     1647..1916,1917..1922,1923..1979,1980..2006,2007..2030,
                     2031..2033,2034..2066,2067..2069,2070..2087,2088..2096))
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6s
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRVIGRLSSLENVYIKADKQKNGIKANFHIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKELFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     primer_bind     1303..1322
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1562..1583
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1689..1710)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      1991..2000
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2093..2102
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2234..2822
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2287..2307)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2705..2716)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2830..3054
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3056..3196
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3056..3185
                     /label=AAV2 ITR
     rep_origin      3271..3726
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3358..3377)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3568..3589
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3743..3762)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3862..3884
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3922..3940)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4008..4112
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4113..4181,4182..4973)
                     /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(4331..4350)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5144..5732
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5633..5652
                     /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 gtcgacgcta 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 gtcgaattcg aagttcctat tctatcagaa gtataggaac ttcgcagaat
      661 ggtagctgga ttgtagctgc tattagcaat atgaaacctc ttagaagttc ctattcttat
      721 aggagtatag gaacttcgct agctcacttc gctgtcatca tttgtacaaa ctcttcgtag
      781 tttacctgac catccccatc gatgtctgct tccctgatca tttcatcaac ctcttcatct
      841 gttaacttct ctccaaggtt tgtcatcacg tggcgaagct ctgctgcact gatgtagcca
      901 ttgccatcct tatcaaacac accgaacgct tctctaattt cttcttccgt gtccctgtat
      961 ttcatttttc ttgccatcat tgtcaggaac tcagggaagt cgattgtgcc gtcaccgtcg
     1021 gcatctactt cattgatcat gtcctgcagc tctgcttctg tggggttctg ccccagagac
     1081 cgcatcaccg tccccagctc cttggttgtt attgtcccat ccccgtcctt gtcaaatagg
     1141 gagaaaagct ctttaaattc tgcgatctgc tcttcagtca gttggtcagg aaggttgtac
     1201 tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt aagctcgatg
     1261 cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg
     1321 tccttgaaga agatggtgcg ctcctggatg tagccttcgg gcatggcgga cttgaagaag
     1381 tcgtgctgct tcatgtggtc ggggtagcgg ctgaagcact gcacgccgta ggtcagggtg
     1441 gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc
     1501 agcttgccgt aggtggcatc gccctcaccc tcgccggaca cgctgaactt gtggccgttt
     1561 acgtcgccgt ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg
     1621 ctcaccatgc tccctccggt accgcccttg tacagctcgt ccatgccgag agtgatcccg
     1681 gcggcggtca cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttcgaa
     1741 agtttggact gcacgctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg
     1801 ggggtgttct gctggtagtg gtaggcgagc tgcacgccgc cgtcctcgat gttgtggcgg
     1861 atgtggaagt tcgccttgat gccgttcttc tgcttgtcgg ccttgatata gacgttctcg
     1921 agtgagctca gccgacctat cactctgact gcgtgacctg tcttattcca cttacgacgt
     1981 gatgagtcga ccatggtggc gagatcctta tcgtcatcgt cgtacagatc ccgacccatt
     2041 tgctgtccac cagtcatgct agccatacca tgatgatgat gatgatgaga acccatggtg
     2101 gcggcgcgcc gaagttccta tacttctgat agaataggaa cttcttgcct taacccagaa
     2161 attatcactg ttattcttta gaatggtgca aagagaagtt cctatactcc tataagaata
     2221 ggaacttctc cggaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     2281 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     2341 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     2401 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     2461 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     2521 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     2581 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     2641 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     2701 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     2761 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     2821 gcctggatcc tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     2881 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     2941 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     3001 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcaggaa
     3061 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     3121 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     3181 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     3241 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg
     3301 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3361 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3421 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3481 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3541 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3601 accctatctc gggctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     3661 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     3721 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     3781 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     3841 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     3901 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     3961 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4021 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4081 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     4141 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     4201 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     4261 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     4321 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     4381 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     4441 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     4501 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     4561 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     4621 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     4681 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     4741 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     4801 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     4861 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     4921 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     4981 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5041 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5101 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5161 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5221 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5281 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5341 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5401 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5461 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5521 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5581 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5641 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5701 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5761 tcctggcctt ttgctggcct tttgctcaca tgt
//
