LOCUS       sequence-212231-        6633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An AAV vector that expresses double floxed cyan pre-eGRASP(p30) 
            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(p30)
FEATURES             Location/Qualifiers
     source          1..6633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1179
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          232..1170
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1127..1147
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1210..1243)
                     /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(1294..1327)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     2547..2568
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2682..2691
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2696..2729
                     /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    2780..2813
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2833..3421
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2886..2906)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3304..3315)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3434..3658
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3666..3806
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3666..3795
                     /label=AAV2 ITR
     rep_origin      3881..4336
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3968..3987)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4178..4199
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4353..4372)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4472..4494
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4532..4550)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4618..4722
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4723..4791,4792..5583)
                     /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(4941..4960)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5754..6342
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6243..6262
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6404..6533
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc
     1201 gactccggaa taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga
     1261 ttgtagctgc tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg
     1321 aagttatggc gcgccttaga cataatactc cttatccttg ttcttcttgt ttttgttggc
     1381 gcttttcgca ctgctgggct gcttctcctt gaccacagcc ccattggact gtgctgagtt
     1441 actgatgtag tttcgactct catccacgtg atatgagcct tcatcccggt ttctgtactt
     1501 gtacatggca tagaggagga tgaggatgca cagagctgct gctgcgacaa tccccaccac
     1561 catgccagtg gtactgctag actcccggat cacctctgcc gagcctgggt atggttcgcg
     1621 gccgcccact cgggtggggt tggctaaccc acctgagctc ggctcacagg ggtcaatgtc
     1681 ctcatcgtcg ctgggacact ctgccgaggc cacaaggata tcatccgtgg tctggctgat
     1741 aggctccttt gtggggggct ttcctcgtcg agctgtgctg gtagccaggg ttgtggtggt
     1801 ctccatgatt gaggtggaca tctcggactg catggcagtg gctgtcgact cagttgtcat
     1861 agaggaaggc acttcggtac cgcttccgcc tccggaacct cctccaggtg gcagtggtgg
     1921 aggctttgta ggagctctag atccgccact gcctccagat ccaccggtct tctcgttggg
     1981 gtctttgctc agcacgctct gggtgctcag gtagtggttg tcgggcagca gcacggggcc
     2041 gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc tgcacgctgc cgtcctccac
     2101 gttgtggcgc acggtgaagt tggccttgat gccgttcttc tgcttgtcgg cggtgatata
     2161 gacgttgccg ctgttgaagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt
     2221 gaagtcggtg cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcaccac
     2281 ggcgcgggtc ttgtacttgc cgtcgtcctt gaaggagatg gtgcgctcct ggacgtagcc
     2341 ttcgggcatg gcggacttga agaagtcgtg ccgcttcatg tggtcggggt agcgggcgaa
     2401 gcactgcacg ccccagctca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc
     2461 ggtggtgcag atgaacttca gggtcagctt gccgatggtg gcatcgccct cgccctcgcc
     2521 cctcacgctg aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac
     2581 cccggtgaac agctcctcgc ccttgctgcc tccgaccggt gcgtcgccgg tgctgccggg
     2641 cacccagagc agcagcaccc atagcaggag ggtgtcggtc tccatggtgg ctagaataac
     2701 ttcgtataaa gtatcctata cgaagttatt tgccttaacc cagaaattat cactgttatt
     2761 ctttagaatg gtgcaaagaa taacttcgta taatgtatgc tatacgaagt tatgaattcc
     2821 tgcagccccg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     2881 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     2941 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     3001 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     3061 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     3121 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     3181 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     3241 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     3301 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     3361 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     3421 catcgggggg atcctgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     3481 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     3541 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     3601 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggcg
     3661 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     3721 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     3781 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct
     3841 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
     3901 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta
     3961 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     4021 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     4081 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     4141 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4201 acaacactca actctatctc gggctattct tttgatttat aagggatttt gccgatttcg
     4261 gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     4321 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     4381 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     4441 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     4501 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     4561 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     4621 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4681 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     4741 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     4801 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     4861 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     4921 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     4981 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     5041 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     5101 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     5161 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     5221 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     5281 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     5341 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     5401 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     5461 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     5521 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     5581 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     5641 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5701 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5761 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5821 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5881 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     5941 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6001 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6061 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6121 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6181 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6241 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6301 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6361 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtcctgcag gcagctgcgc
     6421 gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc tttggtcgcc cggcctcagt
     6481 gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt cctgcggccg
     6541 cacgcgtaag ctttgcaaag atggataaag ttttaaacag agaggaatct ttgcagctaa
     6601 tggaccttct aggtcttgaa aggagtggga att
//
