LOCUS       sequence-209575-        6643 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 6643)
  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 6643)
  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..6643
                     /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        242..1419
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          473..1410
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1367..1387
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1450..1483)
                     /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(1534..1567)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     2787..2808
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2922..2931
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2936..2969
                     /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    3020..3053
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3073..3661
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3126..3146)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3544..3555)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3674..3898
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3906..4046
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3906..4035
                     /label=AAV2 ITR
     rep_origin      4121..4576
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4208..4227)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4418..4439
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4593..4612)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4712..4734
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4772..4790)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4858..4962
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4963..5031,5032..5823)
                     /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(5181..5200)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5994..6582
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6483..6502
                     /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 cgcgtaagct ttgcaaagat ggataaagtt
      181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat
      241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
      301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
      361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
      421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
      481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
      541 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc
     1441 gactccggaa taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga
     1501 ttgtagctgc tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg
     1561 aagttatggc gcgccttaga cataatactc cttatccttg ttcttcttgt ttttgttggc
     1621 gcttttcgca ctgctgggct gcttctcctt gaccacagcc ccattggact gtgctgagtt
     1681 actgatgtag tttcgactct catccacgtg atatgagcct tcatcccggt ttctgtactt
     1741 gtacatggca tagaggagga tgaggatgca cagagctgct gctgcgacaa tccccaccac
     1801 catgccagtg gtactgctag actcccggat cacctctgcc gagcctgggt atggttcgcg
     1861 gccgcccact cgggtggggt tggctaaccc acctgagctc ggctcacagg ggtcaatgtc
     1921 ctcatcgtcg ctgggacact ctgccgaggc cacaaggata tcatccgtgg tctggctgat
     1981 aggctccttt gtggggggct ttcctcgtcg agctgtgctg gtagccaggg ttgtggtggt
     2041 ctccatgatt gaggtggaca tctcggactg catggcagtg gctgtcgact cagttgtcat
     2101 agaggaaggc acttcggtac cgcttccgcc tccggaacct cctccaggtg gaggtggtgg
     2161 agagtagcta ggagatctag atccgccact gcctccagat ccaccggtct tctcgttggg
     2221 gtctttgctc agcacgctct gggtgctcag gtagtggttg tcgggcagca gcacggggcc
     2281 gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc tgcacgctgc cgtcctccac
     2341 gttgtggcgc acggtgaagt tggccttgat gccgttcttc tgcttgtcgg cggtgatata
     2401 gacgttgccg ctgttgaagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt
     2461 gaagtcggtg cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcaccac
     2521 ggcgcgggtc ttgtacttgc cgtcgtcctt gaaggagatg gtgcgctcct ggacgtagcc
     2581 ttcgggcatg gcggacttga agaagtcgtg ccgcttcatg tggtcggggt agcgggcgaa
     2641 gcactgcacg ccccagctca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc
     2701 ggtggtgcag atgaacttca gggtcagctt gccgatggtg gcatcgccct cgccctcgcc
     2761 cctcacgctg aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac
     2821 cccggtgaac agctcctcgc ccttgctgcc tccgaccggt gcgtcgccgg tgctgccggg
     2881 cacccagagc agcagcaccc atagcaggag ggtgtcggtc tccatggtgg ctagaataac
     2941 ttcgtataaa gtatcctata cgaagttatt tgccttaacc cagaaattat cactgttatt
     3001 ctttagaatg gtgcaaagaa taacttcgta taatgtatgc tatacgaagt tatgaattcc
     3061 tgcagccccg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     3121 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     3181 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     3241 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     3301 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     3361 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     3421 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     3481 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     3541 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     3601 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     3661 catcgggggg atcctgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     3721 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     3781 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     3841 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggcg
     3901 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     3961 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     4021 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct
     4081 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
     4141 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     4201 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     4261 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     4321 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     4381 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4441 acaacactca accctatctc gggctattct tttgatttat aagggatttt gccgatttcg
     4501 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     4561 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     4621 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     4681 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     4741 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     4801 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     4861 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4921 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     4981 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     5041 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     5101 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     5161 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     5221 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     5281 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     5341 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     5401 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     5461 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     5521 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     5581 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     5641 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     5701 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     5761 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     5821 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     5881 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5941 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     6001 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     6061 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     6121 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     6181 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6241 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6301 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6361 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6421 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6481 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6541 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6601 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt
//
