LOCUS       sequence-489079-        5647 bp    DNA     circular SYN 09-OCT-2025
DEFINITION  5' AAVLINK plasmid for NRXN1 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5647)
  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  JOURNAL   Unpublished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 5647)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 9, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pAAVLINK-EF1alpha-5'-NRXN1
FEATURES             Location/Qualifiers
     source          1..5647
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   94..234
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   105..234
                     /label=AAV2 ITR
     regulatory      2428..2437
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     LTR             2465..2733
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(2746..2957)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     repeat_region   3014..3143
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3205..3793)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3285..3304)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3964..4755,4756..4824))
                     /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     4587..4606
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4825..4929)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4997..5015
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5053..5075)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5175..5194
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(5211..5647,1..19))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5348..5369)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5560..5579
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gcgccgctac agggcgcgta ctatggttgc tttgacgtat gcggtgtgaa ataccgcaca
       61 gatgcgtaag gagaaaatac cgcatcaggc gcccctgcag gcagctgcgc gctcgctcgc
      121 tcactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag
      181 tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctgcggcc
      241 gctcggtccg cacgtgccac accggtctcc ataacttaat aacttcgtat aaagtatcct
      301 atacgaataa tagcttattg gtctccttaa acctgtcttg taaccttgat acttacctgc
      361 acatgccatt gtttttgcaa gggttgctaa ggcacggttt tgctgtttcc tttgagcagg
      421 aaggcttcac tccagcagta ctttgaactt cagccatttg ccggatatct ttgctttggc
      481 catcgatgaa caaatccctg atgcagccca cgtagccata gttgagcaga gcagtccaca
      541 cctcggtggg gaagacaagg ccagctttat tttctggcag cccccccagg tacaactcat
      601 catccaggtc cagaatctca ctctcaccag gagcagtgta gggagtacgc aacgtgttga
      661 cagaaatggt acctgaccgt ccgtctctct ggaagtccac atgataccat tctccatcat
      721 tcactttctt caacagggct tttattttta tagtacctga ccccatgtcc aggaggaggt
      781 agaggtggcc atctagcatc tcaatagcaa agaagtccac ctttatcatc tgtgggtgct
      841 tggcatcttt ctgatgtctt ggcttgccat ggctaaataa gatgaggcca tttggctctg
      901 ttgtacggaa atcaaatgat atggagccag ttttctttgc attccattta ggcaaagaga
      961 tgaaagactc tggggtttca aaggtgattg ggtctaaagt tgcaacattc tcacatttaa
     1021 atgccaccac tccatggatc ttcatcttag gatctccttg cttggcaagt cgagataatt
     1081 ccagcctcac atcattattt ttatatacaa cctctttgag acagcccata aagttgttac
     1141 tgactggtga ccctggaagg tcggctgtgc tgggactgcc tccaacatag aaaaagtcat
     1201 cagaccccag catggtataa tcttcttgcg tgtagcccgt tgtggtaaga atcccatcca
     1261 ctgatattgt caccatagcg tgtccaatgc ctgagtgctg acgcagattc ctggtgactt
     1321 tcacatcatg ccaggcatta tcattaaact ttccattcac aggctccact agtgcttcaa
     1381 aggcccctga tcccaaatta atgaccagag agacagctcc atttttcagg gcaagattga
     1441 cataatcagc cgatttccca gtgtgaagca tcagtccatt cctctgaagg gttttaaatg
     1501 acagagttat ttcatcactg ctgctttgaa tggggttttg agacaagtcg tagcagaagt
     1561 attcagatcc tttgaacgtg gcaatatatt cttcttttcc tttactttta ccttggtcgc
     1621 ccatcatcag gtgcgccaga ccttccacat tgttgtcttc ttggctgcag tccttgccgc
     1681 ggaagccggt tcgcgagcag tcgcacacgg cctggtcgtc caccacggag cacacacctc
     1741 cgttgaggca caccccgccc tcgccctcct cgcccgcctc gcacgggctt cccccgccgc
     1801 tgttgggcgg ctcatcgtcc agcttcacct cgccgctgtc cacgggcagg acctgcgagg
     1861 agttgaccct cacgtcacga atccacccct tgaagggctc ccgctccctc accgaggcca
     1921 gggtgagctt gagcgccgcg gcgcgcagtt ccgggggcag ccccccgacg aaaaggccgc
     1981 tgaacaccgt catgtccctg cgcttggact tgacctccac ccacttggcc tccacctggt
     2041 cgatgaagag cgtggtgttg cggaactggc ggcggatgcg cacgctgtgc caggcgccgt
     2101 cgttaaccgg cgtgtcggcc aggagcgtcg caggctcagc gcagaagatg gagaagctga
     2161 gctgcaggcg gccgccgcgc gtcagaatca gctccaggaa gtcgcagaag ccctcgtcgt
     2221 cgaagtagag cacgaggccg cgggcgctgc gagtcttgag ctggaagctc atctcgctct
     2281 cgcagcaggc gttccacttg gggaagcgcg tccattggcc ctcggcgccc ggaaactcca
     2341 gcccgctgcc cagctccgcc cagcagccca ggagcagcag cgagaggcac agaagaaaac
     2401 agcccccgcg ctggagcagc gccgtcccca tggtggccgt acggtaggcg ccggtcacag
     2461 cttggatctg taacggcgca gaacagaaaa cgaaacaaag acgtagagtt gagcaagcag
     2521 ggtcaggcaa agcgtggaga gccggctgag tctaggtagg ctccaaggga gcgccggaca
     2581 aaggcccggt ctcgacctga gctttaaact tacctagacg gcggacgcag ttcaggaggc
     2641 accacaggcg ggaggcggca gaacgcgact caaccggcgt ggatggcggc ctcaggtagg
     2701 gcggcgggcg cgtgaaggag agatgcgagc ccctcgaagc ttcagctgtg ttctggcggc
     2761 aaacccgttg cgaaaaagaa cgttcacggc gactactgca cttatatacg gttctccccc
     2821 accctcggga aaaaggcgga gccagtacac gacatcactt tcccagttta ccccgcgcca
     2881 ccttctctag gcacccgttc aattgccgac ccctcccccc aacttctcgg ggactgtggg
     2941 cgatgtgcgc tctgcccact gacgggcacc ggagcgatcg cagatcctct agagtcgaca
     3001 cgcgtgcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     3061 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggcggcct cagtgagcga
     3121 gcgagcgcgc agctgcctgc aggacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3181 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3241 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3301 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3361 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3421 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3481 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3541 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3601 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     3661 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3721 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3781 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     3841 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     3901 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     3961 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4021 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4081 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4141 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4201 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4261 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4321 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4381 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     4441 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     4501 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4561 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     4621 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     4681 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     4741 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     4801 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     4861 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     4921 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     4981 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     5041 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     5101 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     5161 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata aaattgtaaa
     5221 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     5281 atagaccgaa atcggcaaaa tcccttataa atcaaaagaa tagcccgaga tagagttgag
     5341 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     5401 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccca aatcaagttt
     5461 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     5521 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     5581 gggcgctaag gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     5641 gcttaat
//
