LOCUS       sequence-476351-        7021 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the hyaluronan-binding domain of Neurocan fused to eGFP 
            under the CMV promoter. It contains a sequence for secretory 
            pathway, so the probe is exported to the extracellular space.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7021)
  AUTHORS   Fernandez-Ballester M, Ardaya M, Dutheil N, Largitte L, Martin A, 
            Soria FN
  TITLE     Viral-mediated fluorescent labeling of hyaluronan reveals 
            extracellular matrix dynamics in the mouse brain in vivo
  JOURNAL   bioRxiv 2025.01.31.635882; doi: 10.1101/2025.01.31.635882
REFERENCE   2  (bases 1 to 7021)
  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"; journalName: "bioRxiv 
            2025.01.31.635882; doi: 10.1101/2025.01.31.635882"
COMMENT     Sequence Label: pAAV-CMVie-NcanGFP
FEATURES             Location/Qualifiers
     source          1..7021
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        217..1201
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1236..1254
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1236..1253
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     sig_peptide     1304..1363
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             1388..1405
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(2445..2469)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      2468..2477
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2474..2476,2477..2479,2480..3193)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2474..2476,2477..2479,2480..3190)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2519..2540)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2780..2799)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3127..3148
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3285..3305)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3703..3714)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3781..3798)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3787..3994
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4082..4211
                     /label=AAV2 ITR
     primer_bind     complement(4230..4247)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4230..4246)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4239..4261)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4388..4843
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4475..4494)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4685..4706
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4869..4973
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4974..5042,5043..5834)
                     /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(5192..5211)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6005..6593
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6494..6513
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6881..6902
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6917..6922,6923..6940,6941..6947)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6955..6971
                     /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     6960..6982
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6979..6995
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6979..6995
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ccttaattag gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc ttgtagttaa tgattaaccc gccatgctac ttatctacgt
      181 agccatgctc taggaagatc ggaattcgcc cttaaggtcg accgccaatt caatatggcg
      241 tatatggact catgccaatt caatatggtg gatctggacc tgtgccaatt caatatggcg
      301 tatatggact cgtgccaatt caatatggtg gatctggacc ccagccaatt caatatggcg
      361 gacttggcac catgccaatt caatatggcg gacctggcac tgtgccaact ggggaggggt
      421 ctacttggca cggtgccaag tttgaggagg ggtcttggcc ctgtgccaag tccgccatat
      481 tgaattggca tggtgccaat aatggcggcc atattggcta tatgccagga tcaatatata
      541 ggcaatatcc aatatggccc tatgccaata tggctattgg ccaggttcaa tactatgtat
      601 tggccctatg ccatatagta ttccatatat gggttttcct attgacgtag atagcccctc
      661 ccaatgggcg gtcccatata ccatatatgg ggcttcctaa taccgcccat agccactccc
      721 ccattgacgt caatggtctc tatatatggt ctttcctatt gacgtcatat gggcggtcct
      781 attgacgtat atggcgcctc ccccattgac gtcaattacg gtaaatggcc cgcctggctc
      841 aatgcccatt gacgtcaata ggaccaccca ccattgacgt caatgggatg gctcattgcc
      901 cattcatatc cgttctcacg ccccctattg acgtcaatga cggtaaatgg cccacttggc
      961 agtacatcaa tatctattaa tagtaacttg gcaagtacat tactattgga agtacgccag
     1021 ggtacattgg cagtactccc attgacgtca atggcggtaa atggcccgcg atggctgcca
     1081 agtacatccc cattgacgtc aatggggagg ggcaatgacg caaatgggcg ttccattgac
     1141 gtaaatgggc ggtaggcgtg cctaatggga ggtctatata agcaatgctc gtttagggaa
     1201 ccgccattct gcctggggac gtcggagcaa gcttgattta ggtgacacta tagaatacaa
     1261 gctacttgtt ctttttgcag gatcccatcg attcgaattc accatggaga cagacacact
     1321 cctgctatgg gtactgctgc tctgggttcc aggttccact ggtgacgctc cactgggaag
     1381 aggatctcac caccaccacc accacggagg actggctgat caggacacac aggacaccac
     1441 cgccacggaa aaggggcttc gcatgctgaa gtcagggtca ggacccgtcc gtgctgccct
     1501 ggcagagcta gtggccctgc cctgcttctt taccctgcaa ccacggctaa gctccctgcg
     1561 agacattcct cggatcaagt ggactaaggt tcagactgca tcaggccagc gacaggattt
     1621 gccaatcttg gtggccaaag acaacgtggt gcgtgtggcc aagggctggc agggacgggt
     1681 gtcattgcct gcttatcccc ggcacagagc caatgctacc cttctgctgg ggccacttcg
     1741 agcaagcgac tctgggctgt atcgctgcca agtggtaaag ggtatcgaag atgagcagga
     1801 cctggtaacc ctggaagtga caggcgtcgt gttccattat cgggcggccc gggaccgcta
     1861 tgcgctgacc ttcgctgagg cccaggaggc ttgtcgccta agctctgcta ccatcgctgc
     1921 cccacggcac ctgcaggctg cctttgaaga tggctttgac aactgcgacg cgggctggct
     1981 ctcagaccgc acggttcggt acccgatcac tcagtcgcgt cctggttgct atggtgaccg
     2041 cagcagtctc ccgggtgttc ggagctacgg gagacgcgac ccgcaggaac tctacgatgt
     2101 ctactgcttt gcccgcgagc tagggggtga agtcttttac gtgggcccgg cccgccgact
     2161 gaccctggcg ggcgcgcggg cacaatgtca gcggcagggt gcagcgctgg cctcggtggg
     2221 gcagttgcac ctggcctggc acgagggcct ggaccagtgc gacccgggct ggctggcaga
     2281 cggcagcgtg cgctacccca tccagactcc gcgccggcgt tgcgggggcc ccgccccagg
     2341 tgtgcgcact gtgtaccgct tcgccaaccg caccggcttt cctgcgccag gagcacgctt
     2401 cgacgcctac tgcttccgag ctcatcacgg atcttctgga gcttacccag ctttcttgta
     2461 caaagtggcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     2521 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     2581 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     2641 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     2701 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     2761 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     2821 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     2881 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     2941 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     3001 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     3061 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     3121 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     3181 gctgtacaag taggaattca aggcctctcg agcctctaga acgcgtgatc caatcaacct
     3241 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     3301 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     3361 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     3421 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     3481 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     3541 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     3601 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
     3661 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     3721 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgagatctg
     3781 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     3841 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     3901 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     3961 aggattggga agacaatagc aggcatgctg gggactcgag ttaagggcga attcccgata
     4021 aggatcttcc tagagcatgg ctacgtagat aagtagcatg gcgggttaat cattaactac
     4081 aaggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     4141 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     4201 cgagcgcgca gccttaatta acctaattca ctggccgtcg ttttacaacg tcgtgactgg
     4261 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     4321 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     4381 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4441 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     4501 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     4561 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     4621 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4681 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     4741 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     4801 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
     4861 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4921 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     4981 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     5041 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     5101 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     5161 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     5221 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5281 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5341 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     5401 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5461 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     5521 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     5581 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     5641 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     5701 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     5761 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     5821 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     5881 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5941 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6001 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6061 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6121 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     6181 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6241 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6301 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6361 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6421 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6481 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6541 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6601 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     6661 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     6721 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     6781 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     6841 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     6901 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     6961 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaga tttaattaag
     7021 g
//
