LOCUS       sequence-489078-        6488 bp    DNA     circular SYN 09-OCT-2025
DEFINITION  5' AAVLINK plasmid for NRXN2 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6488)
  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  JOURNAL   Unpublished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 6488)
  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            "Unpublished"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'-NRXN2
FEATURES             Location/Qualifiers
     source          1..6488
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      2090..2099
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     LTR             2127..2395
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(2408..2619)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     repeat_region   2676..2805
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2867..3455)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2947..2966)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3626..4417,4418..4486))
                     /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     4249..4268
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4487..4591)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4659..4677
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4715..4737)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4837..4856
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4873..5328)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5010..5031)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5222..5241
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5403..5543
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5414..5543
                     /label=AAV2 ITR
ORIGIN
        1 ggggcagatg cacactgctt cagcgtctcc cgggagcaaa agggggcaac gcccacagcc
       61 ccctgagcct cagccaggcc ccggaggtct cggctacgcc catctatgaa gaggtcccgc
      121 acacagccca cgtagcctgc ccggagtgct gctgtccaca cctctggggg caggggcagg
      181 tccacccggc ccccctcagg gagaccgccc aggtacagct cactctccag gtccagaatc
      241 tcgctgtctc cagtggccaa gaacggcgtg ctgcgactat tcactgagat ggagcctttt
      301 cgcccatccc tctggaagtc cacgtgacac cactcgccat cattgacctt gcggctggat
      361 gcccgcagct tgatgccccc agatcccatg tccagcagaa gatagaggtg gccgtccaat
      421 agctccatgg caaagtagtc ggcccgctga gcagagctgt ggctgccagc tccaccccca
      481 gcccgccggc cctggctgaa gagcagcagc ccattgggct cggtggtgcg gaagtctagg
      541 gagatggagc cagtgcgctt agcgctccag cggggcagcg ccacaaaggc ctcgggactc
      601 tcaaaggtca cagggtccag ggcagccaca tcctcacagc ggaatgacaa gtccccctgc
      661 agcttcatct tggggtcccc ttcctttgcc aggcgggata gttccaattt gaagtcattg
      721 ttcttataga ccacgtcctt gaggcagccc atgaagttgt tgctgacggg cgagcccggc
      781 aggtcagctg tgttggggct gcccccaatg tagaagaagt catcagagcc cagcatggtg
      841 taatcctcct gcgtgtagcc tgtggtggtc aggatcccgt ccaccgagat ggtcaccaga
      901 taatgcagtt tgtttaccat agcgtgtcca atccctgcgt gctggcgcag gtttcgggtg
      961 acccggacgt cgtgccaggc gttgtcgttg aacttgccat tgacgggttc cacaagggcc
     1021 tcgaaggcac ctgagcctag gttgatgacc agccagacag ccccagactt gagggacagg
     1081 ttgacgtagt cggccgactt gcctgtatgc agcatcaggc cgttgcgttg cagggtgcgg
     1141 aaggccagtg tgatctcatc agtgctgctc tggatggggt tgtgtgacag gtcgtagcag
     1201 aagaactcat tgcctttgaa ggtcgccaca aactcctcct tgccttttgt tggctggtgc
     1261 acatcgccgg ctcctcctct cccggccccc ccctcggaga acagtaagga ccctacttcg
     1321 ctgtttaacg tcaggtgagc cggaccttcc atggggtgct cctcttcgct gcagaacttg
     1381 ccgccgaagc ccgtgtggct gcagtcgcag cccacctcgc cgggggccag cacggtgcag
     1441 aggccgccgt tggcgcaggg gttgcgcgcg ggcgcgcaca gcgggtcggc ggtggcgccg
     1501 cgcaggccct ggctgcccag cagcgcgggg ggccgctcgc ccagcttcag gttggccaag
     1561 aggccgcgga agggcggctc gtacttgacg gtgctcagcg taagcgccga gaggcgcacg
     1621 tcgggcggga tgccgcccac gaacaggtcg ctggccacct gcatctcgcg ccgcttggag
     1681 cgcacctcgg cggcgcgggc ctcgccgtcc accgccagcg ccgtgcggcg cgcgtcgcgg
     1741 gtcagcagca ccatgtgcca gcggtcgtcg gccaccggcg tgtccagctg cagcgtggcc
     1801 ggctcggcgc acgaaagcgt gaagcgcagc cgcaggcggc cgtccaccag cagcagctcc
     1861 aggaagtcgc agtcgccgcc gtcgtccagg tagagcagca gcgcgcgcgt ggcgttggtg
     1921 cgcaggctga agctgagctc gccgctgctc gccgcgcccg cccagcgcgc gtagcgagcc
     1981 cactgcccgg ggccgccgcc gaactccagg ccgtccgcgc gcgccgccag cgccagcagc
     2041 agcagcaaca gcagcggcgg cggtgtcggc cgccaccggc tcccggacgc catggtggcc
     2101 gtacggtagg cgccggtcac agcttggatc tgtaacggcg cagaacagaa aacgaaacaa
     2161 agacgtagag ttgagcaagc agggtcaggc aaagcgtgga gagccggctg agtctaggta
     2221 ggctccaagg gagcgccgga caaaggcccg gtctcgacct gagctttaaa cttacctaga
     2281 cggcggacgc agttcaggag gcaccacagg cgggaggcgg cagaacgcga ctcaaccggc
     2341 gtggatggcg gcctcaggta gggcggcggg cgcgtgaagg agagatgcga gcccctcgaa
     2401 gcttcagctg tgttctggcg gcaaacccgt tgcgaaaaag aacgttcacg gcgactactg
     2461 cacttatata cggttctccc ccaccctcgg gaaaaaggcg gagccagtac acgacatcac
     2521 tttcccagtt taccccgcgc caccttctct aggcacccgt tcaattgccg acccctcccc
     2581 ccaacttctc ggggactgtg ggcgatgtgc gctctgccca ctgacgggca ccggagcgat
     2641 cgcagatcct ctagagtcga cacgcgtgcg gccgcaggaa cccctagtga tggagttggc
     2701 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     2761 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggacatg tgagcaaaag
     2821 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2881 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2941 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3001 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3061 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3121 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3181 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3241 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3301 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3361 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3421 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3481 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3541 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3601 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     3661 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     3721 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     3781 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     3841 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     3901 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     3961 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     4021 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     4081 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4141 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4201 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4261 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     4321 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     4381 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     4441 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     4501 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4561 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     4621 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     4681 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     4741 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     4801 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     4861 gagtgcacca taaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt
     4921 aaatcagctc attttttaac caatagaccg aaatcggcaa aatcccttat aaatcaaaag
     4981 aatagcccga gatagagttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     5041 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     5101 aaccatcacc caaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     5161 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     5221 aagggaagaa agcgaaagga gcgggcgcta aggcgctggc aagtgtagcg gtcacgctgc
     5281 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtac tatggttgct
     5341 ttgacgtatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     5401 cccctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg
     5461 tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc
     5521 caactccatc actaggggtt cctgcggccg ctcggtccgc acgtgccaca ccggtctcca
     5581 taacttaata acttcgtata aagtatccta tacgaataat agcttattgg tctccttaaa
     5641 cctgtcttgt aaccttgata cttaccttga ccagctcgat gacaatgaag tcattgccgt
     5701 tgcccgagtt gaacagaaga agcccatcag gggccgtggt cttgaactgg aagaagaggt
     5761 gcatggaagc ataggcttgg agcgtggcga gtgccaggta gctgctgcga ctcttgaagg
     5821 tgacgggatc ggccacgatg gcacgcaggc caaagcgagc attgagctca cagtaggtga
     5881 tgtcaccatc cttgcactgg tccatgtagg gctggccatt gaacacgagc ccactcagat
     5941 gcccgatgaa gttggagggc accacggaga taaaccgccg ctccgtcatg atgcccgtct
     6001 caatgttgtg gaactccagc cgcatatggg ctcctgccat ctgtccctcc acagtcacgt
     6061 tgtccacaga cagctgcagg ctcttgccac gccggaccac cctcaccgtg tgccactcat
     6121 tgtcattgag cttgtgcccc gcaaacagcg tttcggggcc tttactgggt gcgcagccga
     6181 cgcgcaggca gtcgaggttg acagtgagct tcatctgccc cccatccagc tccaggcgta
     6241 gggtgtcggc agactccctg gaagtggtgg ccatcatgag tccgtaggcc cgctgggaca
     6301 tgaaacgcag ggacacatcc tctgcctccg tgtgcatggc gttaggcagc atgatcttca
     6361 tgtacatgga gccatcgtag ctcaggaccg tggcctctct ctcacagacc cgcccaagaa
     6421 agccggtccc gatgcagtca cagatgaagc ggttccagcc ttctcgacag acgcccccat
     6481 tgcgacag
//
