LOCUS       sequence-212227-        5493 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An AAV vector that expresses green pre-eGRASP(p32) under the CaMKIIa
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5493)
  AUTHORS   Choi JH, Sim SE, Kim JI, Choi DI, Oh J, Ye S, Lee J, Kim T, Ko HG, 
            Lim CS, Kaang BK
  TITLE     Interregional synaptic maps among engram cells underlie memory 
            formation.
  JOURNAL   Science. 2018 Apr 27;360(6387):430-435. doi: 
            10.1126/science.aas9204.
  PUBMED    29700265
REFERENCE   2  (bases 1 to 5493)
  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: "10.1126/science"; 
            journalName: "Science"; date: "2018-04-27- 27"; volume: "360"; 
            issue: "6387"; pages: "430-435"
COMMENT     Sequence Label: pAAV-CWB-pre-eGRASP(p32)
FEATURES             Location/Qualifiers
     source          1..5493
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             79..537
                     /codon_start=1
                     /product="improved N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN155(I152L)
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATIGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKRHDFFKSAMPEGYVQERTISFKDDGK
                     YKTRAVVKFEGDTLVNRIELKGTDFKEDGNILGHKLEYNFNSHNVYITA"
     primer_bind     complement(118..139)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    1368..1956
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1421..1441)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1839..1850)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1969..2193
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2201..2341
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2201..2330
                     /label=AAV2 ITR
     rep_origin      2416..2871
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2503..2522)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2713..2734
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2888..2907)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3007..3029
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3067..3085)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3153..3257
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3258..3326,3327..4118)
                     /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(3476..3495)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4289..4877
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4778..4797
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4939..5068
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      join(5488..5493,1..4)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atggagaccg acaccctcct gctatgggtg ctgctgctct gggtgcccgg cagcaccggc
       61 gacgcaccgg tcggaggcag caagggcgag gagctgttca ccggggtggt gcccatcctg
      121 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgaggggcga gggcgagggc
      181 gatgccacca tcggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      241 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcgc ccgctacccc
      301 gaccacatga agcggcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      361 cgcaccatct ccttcaagga cgacggcaag tacaagaccc gcgccgtggt gaagttcgag
      421 ggcgacaccc tggtgaaccg catcgagctg aagggcaccg acttcaagga ggacggcaac
      481 atcctggggc acaagctgga gtacaacttc aacagccaca acgtctatat caccgccgac
      541 aagcagaaga acggcatcaa ggccaacttc accgtgcgcc acaacgtgga ggacggcagc
      601 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
      661 cccgacaacc actacctgag cacccagacc gtgctgagca aagaccccaa cgagaagacc
      721 ggtggatctg gaggcagtgg cggatctaga tctcctagct actctccacc acctccacct
      781 ggaggaggtt ccggaggcgg aagcggtacc gaagtgcctt cctctatgac aactgagtcg
      841 acagccactg ccatgcagtc cgagatgtcc acctcaatca tggagaccac cacaaccctg
      901 gctaccagca cagctcgacg aggaaagccc cccacaaagg agcctatcag ccagaccacg
      961 gatgatatcc ttgtggcctc ggcagagtgt cccagcgacg atgaggacat tgacccctgt
     1021 gagccgagct caggtgggtt agccaacccc acccgagtgg gcggccgcga accataccca
     1081 ggctcggcag aggtgatccg ggagtctagc agtaccactg gcatggtggt ggggattgtc
     1141 gcagcagcag ctctgtgcat cctcatcctc ctctatgcca tgtacaagta cagaaaccgg
     1201 gatgaaggct catatcacgt ggatgagagt cgaaactaca tcagtaactc agcacagtcc
     1261 aatggggctg tggtcaagga gaagcagccc agcagtgcga aaagcgccaa caaaaacaag
     1321 aagaacaagg ataaggagta ttatgtctga attcctgcag ccccgataat caacctctgg
     1381 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     1441 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     1501 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     1561 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     1621 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     1681 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     1741 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca
     1801 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     1861 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     1921 agacgagtcg gatctccctt tgggccgcct ccccgcatcg gggggatcct gtgccttcta
     1981 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     2041 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     2101 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     2161 gcaggcatgc tggggatgcg gtgggctcta tggcggccgc aggaacccct agtgatggag
     2221 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     2281 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     2341 gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
     2401 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     2461 cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc
     2521 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     2581 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     2641 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     2701 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct
     2761 attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga
     2821 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
     2881 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     2941 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     3001 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     3061 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     3121 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3181 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3241 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     3301 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     3361 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     3421 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     3481 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     3541 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     3601 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     3661 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     3721 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     3781 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     3841 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     3901 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     3961 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     4021 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     4081 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     4141 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4201 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4261 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4321 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4381 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     4441 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4501 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4561 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4621 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4681 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4741 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4801 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4861 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4921 ggccttttgc tcacatgtcc tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc
     4981 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
     5041 gtggccaact ccatcactag gggttcctgc ggccgcagat gctgcacact tgtggactaa
     5101 gtttgttcgc atccccttct ccaaccccct cagtacatca ccctggggga acagggtcca
     5161 cttgctcctg ggcccacaca gtcctgcagt attgtgtata taaggccagg gcaaagagga
     5221 gcaggtttta aagtgaaagg caggcaggtg ttggggaggc agttaccggg gcaacgggaa
     5281 cagggcgttt cggaggtggt tgccatgggg acctggatgc tgacgaaggc tcgcgaggct
     5341 gtgagcagcc acagtgccct gctcagaagc cccaagctcg tcagtcaagc cggttctccg
     5401 tttgcactca ggagcacggg caggcgagtg gcccctagtt ctgggggcag cgtgaaaaga
     5461 cggtatcgat aagcttgatt cgagctagcc acc
//
