LOCUS       sequence-479557-        6504 bp    DNA     circular SYN 24-JUL-2025
DEFINITION  3' AAVLINK plasmid for ARHGEF10 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6504)
  AUTHORS   Zhonghua Lu Lab
  TITLE     Zhonghua Lu AAV Plasmids 2025
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6504)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 24, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pAAVLINKv1-pCALM1-Cre-3' ARHGEF10
FEATURES             Location/Qualifiers
     source          1..6504
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             277..1308
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     primer_bind     1464..1483
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             3224..3289
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3354..3374)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   3767..3907
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3767..3896
                     /label=AAV2 ITR
     rep_origin      3982..4437
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4069..4088)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4279..4300
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4454..4473)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4573..4595
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4633..4651)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4719..4823
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4824..4892,4893..5684)
                     /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(5042..5061)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5855..6443
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6344..6363
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtcgctct gcgcgaggcg gctccgccgc ggcggaggga
      181 tacggcgcac catatatata tcgcggggcg cagactcgcg ctccggcagt ggtgctggga
      241 gtgtcgtgga cgccgtgccg ttactcgtac gccaccatgt ccaatttact gaccgtacac
      301 caaaatttgc ctgcattacc ggtcgatgca acgagtgatg aggttcgcaa gaacctgatg
      361 gacatgttca gggatcgcca ggcgttttct gagcatacct ggaaaatgct tctgtccgtt
      421 tgccggtcgt gggcggcatg gtgcaagttg aataaccgga aatggtttcc cgcagaacct
      481 gaagatgttc gcgattatct tctatatctt caggcgcgcg gtctggcagt aaaaaccatc
      541 cagcaacatt tgggccagct aaacatgctt catcgtcggt ccgggctgcc acgaccaagt
      601 gacagcaatg ctgtttcact ggttatgcgg cggatccgaa aagaaaacgt tgatgccggt
      661 gaacgtgcaa aacaggctct agcgttcgaa cgcactgatt tcgaccaggt tcgttcactc
      721 atggaaaata gcgatcgctg ccaggatata cgtaatctgg catttctggg gattgcttat
      781 aacaccctgt tacgtatagc cgaaattgcc aggatcaggg ttaaagatat ctcacgtact
      841 gacggtggga gaatgttaat ccatattggc agaacgaaaa cgctggttag caccgcaggt
      901 gtagagaagg cacttagcct gggggtaact aaactggtcg agcgatggat ttccgtctct
      961 ggtgtagctg atgatccgaa taactacctg ttttgccggg tcagaaaaaa tggtgttgcc
     1021 gcgccatctg ccaccagcca gctatcaact cgcgccctgg aagggatttt tgaagcaact
     1081 catcgattga tttacggcgc taaggatgac tctggtcaga gatacctggc ctggtctgga
     1141 cacagtgccc gtgtcggagc cgcgcgagat atggcccgcg ctggagtttc aataccggag
     1201 atcatgcaag ctggtggctg gaccaatgta aatattgtaa tgaactatat ccgtaacctg
     1261 gatagtgaaa caggggcaat ggtgcgcctg ctggaagatg gcgattaggt aagtatcaag
     1321 gttacaagac aggtttaagg agaccaataa ctagtataac ttcgtatagg atactttata
     1381 gcaatttatg aaactgggct tgtcgagaca gagaagactc ttgcgtttct gataggcacc
     1441 tattggtctt actgacatcc actttgcctt tctctccaca ggtttacaaa ggcttatttt
     1501 gaagaaagaa gatgaaatca gagctgcgga ctgctgcaga attcagttac agcttcccgg
     1561 gaagcaggac aaatctgggc gaccgacgtt ctttacagct gtgttcaata cgttcactcc
     1621 tgccatcaag gagtcctggg tcaacagctt acagatggcc aagctcgccc tagaagagga
     1681 gaaccacatg ggctggttct gtgtggaaga cgatgggaat cacattaaaa aggagaagca
     1741 tcctctcctc gtcggacaca tgcccgtgat ggtggccaag cagcaggagt tcaagattga
     1801 atgtgctgct tataaccctg aaccttacct aaataatgaa agccagccag attcattttc
     1861 cacggcacat ggtttcctgt ggatcggaag ttgcacccat caaatgggtc agattgccat
     1921 cgtctcgttt caaaattcca ctcccaaagt cattgagtgc ttcaacgtgg aatctcgcat
     1981 cctgtgcatg ctgtacgttc ccgtcgagga gaagcgcaga gagcctgggg cacccccgga
     2041 ccccgagacc ccggccgtga gagcttctga tgtccccacg atctgtgtag ggacggagga
     2101 gggaagcatt tccatttata aaagcagtca aggctccaag aaagtgagac ttcagcactt
     2161 tttcactcct gagaagtcca cagtcatgag cctggcttgc acgtctcaga gcctgtacgc
     2221 tggcctggtc aacggggcag tcgccagcta cgccagagcc ccagatggat cctgggattc
     2281 agaacctcaa aaagtgatca agttaggcgt cctaccagtt agaagtctac tcatgatgga
     2341 agacacgttg tgggcggctt ccggaggtca agtcttcatc atcagtgtgg agactcatgc
     2401 tgtagagggt cagctggagg cccaccagga ggaaggcatg gtgatctccc acatggccgt
     2461 gtccggcgtc gggatctgga ttgccttcac ctcagggtcc acgctccgcc tttttcacac
     2521 ggaaactctc aagcacctgc aggacatcaa catcgccacc cctgttcaca acatgctgcc
     2581 agggcaccag cggctgtcgg tgacgagcct gctcgtctgc cacggattgc tgatggtcgg
     2641 caccagcctg ggagtcctcg tggccctgcc ggtcccacgt ctgcaaggga ttcccaaagt
     2701 gaccggaaga ggcatggtct cctaccatgc acacaacagt cctgtcaaat tcatcgtcct
     2761 ggccacggct ctgcacgaga aagacaagga caaatccagg gacagcctgg ctcctggccc
     2821 cgagcctcag gacgaagacc agaaggacgc acttccgagt ggaggagctg gttcatctct
     2881 gagccagggt gaccctgacg cagccatctg gttgggagat tcgctgggat cgatgactca
     2941 gaaaagcgac ctgtcctcct catctgggtc cctgagcttg tctcacggct ccagctctct
     3001 agagcacaga tcagaggaca gcaccatcta tgatctcctg aaggatcctg tctcgctgag
     3061 aagcaaagca cgccgggcca agaaagccaa ggccagctcg gcgctggtgg tctgtggagg
     3121 gcagggccac cgccgggtgc acaggaaggc ccggcagccc caccaggaag agctggcgcc
     3181 gaccgtcatg gtctggcaga tccctctgct gaatatatgt acagactaca aagaccatga
     3241 cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt aactcgagat
     3301 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     3361 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     3421 atggctttca ttttctcctc cttgtataaa tcctggttag ttcttgccac ggcggaactc
     3481 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     3541 gtggtgttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat ctagctttat
     3601 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     3661 taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt
     3721 ttaaagatct ggagaccggt gtggcacgtg cggaccgagc ggccgcagga acccctagtg
     3781 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     3841 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
     3901 ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     3961 atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4021 tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt
     4081 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4141 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     4201 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     4261 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct
     4321 cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg
     4381 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat
     4441 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     4501 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     4561 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     4621 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     4681 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     4741 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4801 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     4861 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     4921 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     4981 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     5041 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     5101 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     5161 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5221 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     5281 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5341 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     5401 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     5461 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     5521 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     5581 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     5641 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     5701 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5761 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5821 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5881 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5941 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6001 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6061 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6121 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6181 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6241 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6301 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6361 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6421 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6481 tttgctggcc ttttgctcac atgt
//
