LOCUS       sequence-491774-        7214 bp    DNA     circular SYN 04-NOV-2025
DEFINITION  3' AAVLINK plasmid for NRXN1 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7214)
  AUTHORS   Zhonghua Lu Lab
  TITLE     AAVLINK: A Potent DNA-Recombination Method for Large Cargo Delivery 
            in Gene Therapy
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7214)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 4, 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            "Unpublished"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'-NRXN1-FLAG
FEATURES             Location/Qualifiers
     source          1..7214
                     /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"
     CDS             3934..3999
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(4064..4084)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   4477..4617
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4477..4606
                     /label=AAV2 ITR
     rep_origin      4692..5147
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4779..4798)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4989..5010
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5164..5183)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5283..5305
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5343..5361)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5429..5533
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5534..5602,5603..6394)
                     /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(5752..5771)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6565..7153
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7054..7073
                     /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 gggatgggtg gaacagatat
     1501 gtctgtgatt gttccggaac aggctatctt ggcaggtcct gtgagagaga ggcaacggtt
     1561 ttgagctatg atgggagcat gtttatgaaa attcagctcc ccgtagtcat gcatacggag
     1621 gctgaggatg tttccttacg gttccgatcc cagcgtgcat atggcattct gatggcaacc
     1681 acttctagag actctgctga caccctccgc ctggagctag acgcaggacg tgtgaaactg
     1741 acggtcaatc tagattgtat caggattaac tgtaattcca gcaaaggtcc cgagactctt
     1801 tttgctggct ataacctcaa tgataacgag tggcacacag tgcgtgtagt tcggcgtgga
     1861 aaaagtttaa agttaacagt ggatgaccaa caggccatga caggtcaaat ggcaggtgat
     1921 catactaggc tggagttcca taacatagag actggcatca tcacagaacg acggtatctt
     1981 tcttctgtcc cctccaactt cattggacac ctgcagagct tgacatttaa tggaatggca
     2041 tacattgacc tgtgtaaaaa tggcgacata gattactgtg agcttaatgc cagatttggc
     2101 ttcaggaaca tcatagcaga tcctgtcacc ttcaagacca aatcgagcta tgttgcctta
     2161 gctaccttgc aagcctacac ttctatgcat ctttttttcc agttcaagac aacatcccta
     2221 gatggattaa ttctatataa cagtggggat ggaaatgact ttattgtggt tgaattagtt
     2281 aaagggtact tacattacgt gtttgatttg ggaaatggtg ctaacctcat caaaggaagc
     2341 tcaaataaac ctctcaatga caatcagtgg cacaacgtga tgatatcaag ggacaccagc
     2401 aacctccaca ctgtaaagat tgacacaaaa atcacaacgc aaatcaccgc cggagccagg
     2461 aacttagacc tcaagagtga cttatatata ggaggagtag ctaaagaaac atacaaatcc
     2521 ttaccaaaac ttgtacatgc caaagaaggc tttcaaggct gcctggcatc agttgattta
     2581 aatggacggc ttccggacct catctccgat gctcttttct gcaacggaca gatcgagaga
     2641 ggatgtgaag ggcccagcac aacctgccaa gaggactcat gttccaatca aggtgtgtgc
     2701 ttgcaacaat gggatggctt cagctgtgac tgtagtatga cttccttcag tggaccactc
     2761 tgcaatgacc ctgggacgac atatatcttt agcaaaggtg gtggacaaat cacgtataag
     2821 tggcctccta atgaccgacc cagtacacga gcagacagac tggccatagg ttttagcact
     2881 gttcagaaag aagccgtatt ggtgcgagtg gacagttctt caggcttggg tgactaccta
     2941 gaactgcata tacaccaggg aaaaattgga gttaagttta atgttgggac agatgacatc
     3001 gccattgaag aatccaatgc aatcattaat gatgggaaat accatgtagt tcgtttcacg
     3061 aggagtggtg gcaatgccac gttgcaggtg gacagctggc cagtgatcga gcgctaccct
     3121 gcaggaaaca atgataacga gcgcctggcg attgctagac agcgaattcc atatcgactt
     3181 ggtcgagtag ttgatgaatg gctactcgac aaagggcgtc agctcacaat cttcaatagc
     3241 caagcaacca taataattgg cgggaaagag cagggccagc ccttccaggg ccagctctct
     3301 gggctgtact acaatggctt gaaagttctg aatatggcag ccgaaaacga tgccaacatc
     3361 gccatagtgg gaaatgtgag actggttggt gaagtgcctt cctctatgac aactgagtca
     3421 acagccactg ccatgcaatc agagatgtcc acatcaatta tggagactac cacgaccctg
     3481 gctactagca cagccagaag aggaaagccc ccgacaaaag aacccattag ccagaccaca
     3541 gatgacatcc ttgtggcctc agcagagtgt cccagcgatg atgaggacat tgacccctgt
     3601 gagccgagct caggtgggtt agccaaccca acccgagcag gcggcagaga gccgtatcca
     3661 ggctcagcag aagtgatccg ggagtccagc agcaccacgg gtatggtcgt tgggatagta
     3721 gccgctgccg ccctgtgcat ccttatcctc ctctatgcca tgtacaagta cagaaaccgg
     3781 gatgaaggct cataccatgt ggacgagagt cgaaactaca tcagtaactc agcacagtcc
     3841 aatggggctg ttgtaaagga gaaacaaccc agcagtgcga aaagctccaa caaaaataag
     3901 aaaaacaagg ataaagagta ttatgtctgt acagactaca aagaccatga cggtgattat
     3961 aaagatcatg acatcgacta caaggatgac gatgacaagt aactcgagat aatcaacctc
     4021 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     4081 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     4141 ttttctcctc cttgtataaa tcctggttag ttcttgccac ggcggaactc atcgccgcct
     4201 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttta
     4261 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat ctagctttat ttgtgaaatt
     4321 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     4381 aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagatct
     4441 ggagaccggt gtggcacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg
     4501 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     4561 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc
     4621 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     4681 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4741 cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc
     4801 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4861 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     4921 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4981 ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc
     5041 ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta
     5101 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     5161 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     5221 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     5281 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     5341 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     5401 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     5461 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     5521 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     5581 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     5641 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     5701 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     5761 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     5821 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     5881 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     5941 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     6001 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     6061 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     6121 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     6181 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     6241 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     6301 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     6361 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     6421 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     6481 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     6541 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     6601 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6661 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     6721 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6781 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6841 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6901 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6961 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     7021 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     7081 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     7141 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     7201 ttttgctcac atgt
//
