LOCUS       sequence-396588-        6647 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses full-length mouse HAPLN1 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 6647)
  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 6647)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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..6647
                     /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(1748..1750,1751..2461)
                     /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(1790..1811)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2051..2070)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2398..2419
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2495..2519)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2523..3111
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2576..2596)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2994..3005)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(3117..3137)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     polyA_signal    3165..3286
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3176..3195)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3230..3249
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3390..3530
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3390..3519
                     /label=AAV2 ITR
     primer_bind     complement(3540..3557)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3540..3556)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3549..3571)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3769..4224
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3856..3875)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4066..4087
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4241..4260)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4360..4382
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4420..4438)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4506..4610
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4611..4679,4680..5471)
                     /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(4829..4848)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5642..6230
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6131..6150
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6518..6539
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6554..6559,6560..6577,6578..6584)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6592..6608
                     /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     6597..6619
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6616..6632
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6616..6632
                     /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 ccccgtctac ttgtggaagc agaacaagcc aaggtcttct ctcaccgagg
      841 tggcaacgtg acactgccat gcaaatttta tcgagaccct acagcatttg gctcaggaat
      901 ccacaaaatc cgcatcaagt ggaccaagct aacttcagat tacctcaggg aagtagatgt
      961 cttcgtttcc atgggctatc acaagaagac ctatggaggc tatcaaggtc gagtgtttct
     1021 gaagggaggc agtgataatg atgcctccct ggtcatcacg gatcttaccc tggaggatta
     1081 tgggagatat aaatgtgagg tgattgaagg gctagaagat gatactgctg tggtggcatt
     1141 ggagttacaa ggtgtggtgt tcccttactt tccacgactg ggacgctaca atcttaactt
     1201 tcacgaggca cgccaggctt gtctggacca ggacgcagtg attgcttcct ttgaccagct
     1261 gtacgatgcc tggcggggtg ggctggactg gtgcaatgct ggctggctca gtgatggatc
     1321 tgtgcagtac ccaatcacca aaccacgaga gccctgcggg ggccaaaaca cggtgcctgg
     1381 agtcaggaac tacgggtttt gggacaagga taaaagcaga tatgacgttt tctgttttac
     1441 atccaacttc aacggccgat tttactacct gatccacccc accaaactca cctacgatga
     1501 ggcggtgcaa gcttgtctca atgacggtgc tcagatcgcg aaagtgggcc agatatttgc
     1561 tgcctggaag cttctgggct atgaccgctg cgatgccggc tggctagcgg atggcagcgt
     1621 ccgctatcct atttctcgac caagaaggcg ctgcagtccg actgaggctg cagtgcgctt
     1681 tgtaggtttc ccagataaaa agcataagct atacggggtc tattgcttca gagcatacaa
     1741 cctcgaggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     1801 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     1861 ctacggcaag ctgaccctga agttcatctc caccaccggc aagctgcccg tgccctggcc
     1921 caccctcgtg accaccctga cctacggcgt gcagatgttc gcccgctacc ccgaccacat
     1981 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     2041 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     2101 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     2161 gcacaagctg gagtacaact acaacagcca caacgtctat atcacggccg acaagcagaa
     2221 gaacggcatc aaggccaact tcaagatccg ccacaacatc gaggacggcg gcgtgcagct
     2281 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     2341 ccactacctg agcacccagt ccaaactgag caaagacccc aacgagaagc gcgatcacat
     2401 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     2461 gtaaggtacc gagctcgtcg accatatgtc tagaacccag ctttcttgta caaagtggcg
     2521 ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     2581 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     2641 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     2701 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     2761 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     2821 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     2881 tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc
     2941 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     3001 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     3061 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg catcggcaac
     3121 tttattatac atagttgatg gccggccgct tcgagcagac atgataagat acattgatga
     3181 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     3241 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     3301 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     3361 cctctacaaa tgtggtaatc gatagatcta ggaaccccta gtgatggagt tggccactcc
     3421 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     3481 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg cagcttggca
     3541 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     3601 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     3661 ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt
     3721 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     3781 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3841 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3901 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3961 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     4021 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4081 aaactggaac aacactcaac cctatctcgg gctattcttt tgatttataa gggattttgc
     4141 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     4201 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     4261 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     4321 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     4381 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     4441 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     4501 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4561 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     4621 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4681 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     4741 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     4801 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     4861 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     4921 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     4981 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     5041 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     5101 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     5161 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     5221 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     5281 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     5341 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     5401 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     5461 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     5521 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     5581 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5641 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5701 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     5761 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     5821 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5881 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     5941 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6001 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6061 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6121 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6181 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6241 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     6301 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     6361 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     6421 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     6481 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     6541 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     6601 gataacaatt tcacacagga aacagctatg accatgatta cgaattg
//
