LOCUS       sequence-394043-        6302 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse HAPLN1 mutant lacking the lectican binding domain 
            and fused with cysteine-free GFP (cfGFP) under synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6302)
  AUTHORS   Ramsaran AI, Wang Y, Golbabaei A, Aleshin S, de Snoo ML, Yeung BA, 
            Rashid AJ, Awasthi A, Lau J, Tran LM, Ko SY, Abegg A, Duan LC, 
            McKenzie C, Gallucci J, Ahmed M, Kaushik R, Dityatev A, Josselyn SA,
            Frankland PW
  TITLE     A shift in the mechanisms controlling hippocampal engram formation 
            during brain maturation.
  JOURNAL   Science. 2023 May 5;380(6644):543-551. doi: 10.1126/science.ade6530.
            Epub 2023 May 4.
  PUBMED    37141366
REFERENCE   2  (bases 1 to 6302)
  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.ade6530"; journalName: "Science"; date: "2023-05-5-
            5"; volume: "380"; issue: "6644"; pages: "543-551"
COMMENT     Sequence Label: AAV-∆Hapln1
FEATURES             Location/Qualifiers
     source          1..6302
                     /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
     protein_bind    148..168
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        190..637
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    637..661
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(1403..1405,1406..2116)
                     /codon_start=1
                     /product="strongly enhanced green fluorescent protein 
                     (Kremers et al., 2007)"
                     /label=SGFP2
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FISTTGKLPVPWPTLVTTLTYGVQMFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1445..1466)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1706..1725)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2053..2074
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2150..2174)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2178..2766
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2231..2251)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2649..2660)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(2772..2792)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     polyA_signal    2820..2941
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2831..2850)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2885..2904
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3045..3185
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3045..3174
                     /label=AAV2 ITR
     primer_bind     complement(3195..3212)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3195..3211)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3204..3226)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3424..3879
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3511..3530)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3721..3742
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3896..3915)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4015..4037
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4075..4093)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4161..4265
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4266..4334,4335..5126)
                     /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(4484..4503)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5297..5885
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5786..5805
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6173..6194
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6209..6214,6215..6232,6233..6239)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6247..6263
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6252..6274
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6271..6287
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6271..6287
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tatcgatcaa ctttgtatag aaaagttgct gcagagggcc
      181 ctgcgtatga gtgcaagtgg gttttaggac caggatgagg cggggtgggg gtgcctacct
      241 gacgaccgac cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc
      301 cctatcagag agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc
      361 agcaccgcgg acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac
      421 tgaaggcgcg ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg
      481 tcgcgtccgc gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg
      541 gcacgggcgc gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt
      601 gggcagcgga ggagtcgtgt cgtgcctgag agcgcagcaa gtttgtacaa aaaagcaggc
      661 tgaattcgcc accatgagaa gtctgcttct cctggtgctg atttcagtct gttgggctga
      721 ccaccacctt tcagacagct acactcctcc agatcaagac agagttattc acatccaagc
      781 agaaaatggc gcattggagt tacaaggtgt ggtgttccct tactttccac gactgggacg
      841 ctacaatctt aactttcacg aggcacgcca ggcttgtctg gaccaggacg cagtgattgc
      901 ttcctttgac cagctgtacg atgcctggcg gggtgggctg gactggtgca atgctggctg
      961 gctcagtgat ggatctgtgc agtacccaat caccaaacca cgagagccct gcgggggcca
     1021 aaacacggtg cctggagtca ggaactacgg gttttgggac aaggataaaa gcagatatga
     1081 cgttttctgt tttacatcca acttcaacgg ccgattttac tacctgatcc accccaccaa
     1141 actcacctac gatgaggcgg tgcaagcttg tctcaatgac ggtgctcaga tcgcgaaagt
     1201 gggccagata tttgctgcct ggaagcttct gggctatgac cgctgcgatg ccggctggct
     1261 agcggatggc agcgtccgct atcctatttc tcgaccaaga aggcgctgca gtccgactga
     1321 ggctgcagtg cgctttgtag gtttcccaga taaaaagcat aagctatacg gggtctattg
     1381 cttcagagca tacaacctcg aggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     1441 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     1501 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctccacca ccggcaagct
     1561 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcaga tgttcgcccg
     1621 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     1681 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     1741 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     1801 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcac
     1861 ggccgacaag cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga
     1921 cggcggcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     1981 gctgctgccc gacaaccact acctgagcac ccagtccaaa ctgagcaaag accccaacga
     2041 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     2101 ggacgagctg tacaagtaag gtaccgagct cgtcgaccat atgtctagaa cccagctttc
     2161 ttgtacaaag tggcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     2221 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     2281 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     2341 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     2401 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     2461 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     2521 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt
     2581 cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     2641 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     2701 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     2761 ccccgcatcg gcaactttat tatacatagt tgatggccgg ccgcttcgag cagacatgat
     2821 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     2881 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     2941 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     3001 ttaaagcaag taaaacctct acaaatgtgg taatcgatag atctaggaac ccctagtgat
     3061 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     3121 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct
     3181 gcaggcagct tggcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     3241 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     3301 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg
     3361 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     3421 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     3481 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     3541 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     3601 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     3661 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     3721 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt
     3781 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     3841 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     3901 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     3961 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     4021 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     4081 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     4141 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4201 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4261 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     4321 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     4381 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     4441 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     4501 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     4561 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     4621 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     4681 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     4741 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     4801 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     4861 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     4921 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     4981 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5041 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5101 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     5161 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5221 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5281 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5341 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5401 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     5461 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5521 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5581 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5641 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5701 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5761 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5821 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5881 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5941 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     6001 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     6061 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     6121 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     6181 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     6241 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgaat
     6301 tg
//
