LOCUS       sequence-212596-        6633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An AAV vector that expresses double floxed cyan pre-eGRASP(p32) 
            under the Ef1a promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6633)
  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 6633)
  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-EWB-DIO-cyan pre-eGRASP(p32)
FEATURES             Location/Qualifiers
     source          1..6633
                     /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        231..1409
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          462..1400
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1357..1377
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1440..1473)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(1524..1557)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     2777..2798
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2912..2921
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2926..2959
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    3010..3043
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3063..3651
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3116..3136)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3534..3545)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3664..3888
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3896..4036
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3896..4025
                     /label=AAV2 ITR
     rep_origin      4111..4566
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4198..4217)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4408..4429
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4583..4602)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4702..4724
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4762..4780)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4848..4952
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4953..5021,5022..5813)
                     /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(5171..5190)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5984..6572
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6473..6492
                     /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 gcgtaagctt tgcaaagatg gataaagttt taaacagaga
      181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
      241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
      541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
      601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
      661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
      721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
      781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
      841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
      901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
      961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     1381 gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc gactccggaa
     1441 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc
     1501 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc
     1561 gcgccttaga cataatactc cttatccttg ttcttcttgt ttttgttggc gcttttcgca
     1621 ctgctgggct gcttctcctt gaccacagcc ccattggact gtgctgagtt actgatgtag
     1681 tttcgactct catccacgtg atatgagcct tcatcccggt ttctgtactt gtacatggca
     1741 tagaggagga tgaggatgca cagagctgct gctgcgacaa tccccaccac catgccagtg
     1801 gtactgctag actcccggat cacctctgcc gagcctgggt atggttcgcg gccgcccact
     1861 cgggtggggt tggctaaccc acctgagctc ggctcacagg ggtcaatgtc ctcatcgtcg
     1921 ctgggacact ctgccgaggc cacaaggata tcatccgtgg tctggctgat aggctccttt
     1981 gtggggggct ttcctcgtcg agctgtgctg gtagccaggg ttgtggtggt ctccatgatt
     2041 gaggtggaca tctcggactg catggcagtg gctgtcgact cagttgtcat agaggaaggc
     2101 acttcggtac cgcttccgcc tccggaacct cctccaggtg gaggtggtgg agagtagcta
     2161 ggagatctag atccgccact gcctccagat ccaccggtct tctcgttggg gtctttgctc
     2221 agcacgctct gggtgctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg
     2281 ggggtgttct gctggtagtg gtcggcgagc tgcacgctgc cgtcctccac gttgtggcgc
     2341 acggtgaagt tggccttgat gccgttcttc tgcttgtcgg cggtgatata gacgttgccg
     2401 ctgttgaagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcggtg
     2461 cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcaccac ggcgcgggtc
     2521 ttgtacttgc cgtcgtcctt gaaggagatg gtgcgctcct ggacgtagcc ttcgggcatg
     2581 gcggacttga agaagtcgtg ccgcttcatg tggtcggggt agcgggcgaa gcactgcacg
     2641 ccccagctca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag
     2701 atgaacttca gggtcagctt gccgatggtg gcatcgccct cgccctcgcc cctcacgctg
     2761 aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac
     2821 agctcctcgc ccttgctgcc tccgaccggt gcgtcgccgg tgctgccggg cacccagagc
     2881 agcagcaccc atagcaggag ggtgtcggtc tccatggtgg ctagaataac ttcgtataaa
     2941 gtatcctata cgaagttatt tgccttaacc cagaaattat cactgttatt ctttagaatg
     3001 gtgcaaagaa taacttcgta taatgtatgc tatacgaagt tatgaattcc tgcagccccg
     3061 ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     3121 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     3181 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     3241 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     3301 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     3361 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     3421 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     3481 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     3541 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     3601 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgggggg
     3661 atcctgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     3721 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     3781 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     3841 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggcg gccgcaggaa
     3901 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     3961 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     4021 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     4081 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg
     4141 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc
     4201 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     4261 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4321 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4381 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     4441 actctatctc gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt
     4501 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     4561 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     4621 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     4681 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     4741 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     4801 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4861 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4921 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     4981 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     5041 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     5101 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     5161 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     5221 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     5281 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     5341 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     5401 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     5461 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     5521 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     5581 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5641 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     5701 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     5761 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     5821 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5881 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5941 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6001 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6061 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6121 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6181 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6241 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6301 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6361 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6421 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6481 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6541 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6601 tcctggcctt ttgctggcct tttgctcaca tgt
//
