LOCUS       sequence-489082-        6077 bp    DNA     circular SYN 09-OCT-2025
DEFINITION  5' AAVLINK plasmid for TRPM1 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6077)
  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  JOURNAL   Unpublished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 6077)
  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            "Unpublished"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-HA-5'-TRPM1
FEATURES             Location/Qualifiers
     source          1..6077
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   2312..2452
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2312..2441
                     /label=AAV2 ITR
     rep_origin      2527..2982
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2614..2633)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2824..2845
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2999..3018)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3118..3140
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3178..3196)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3264..3368
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3369..3437,3438..4229)
                     /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(3587..3606)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4400..4988
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4889..4908
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5050..5179
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        5236..5447
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             5460..5728
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             5765..5791
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
ORIGIN
        1 atatccgtgt atcctatgac accaagccag actcactgct ccatctcatg gtgaaagatt
       61 ggcagctgga actccccaag ctcttaatat ctgtgcatgg aggcctccag aactttgaga
      121 tgcagcccaa gctgaaacaa gtctttggga aaggcctgat caaggctgct atgaccaccg
      181 gggcctggat cttcaccggg ggtgtcagca caggtgttat cagccacgta ggggatgcct
      241 tgaaagacca ctcctccaag tccagaggcc gggtttgtgc tataggaatt gctccatggg
      301 gcatcgtgga gaataaggaa gacctggttg gaaaggatgt aacaagagtg taccagacca
      361 tgtccaaccc tctaagtaag ctctctgtgc tcaacaactc ccacacccac ttcatcctgg
      421 ctgacaatgg caccctgggc aagtatggcg ccgaggtgaa gctgcgaagg ctgctggaaa
      481 agcacatctc cctgcagaag atcaacacaa gactggggca gggcgtgccc ctcgtgggtc
      541 tcgtggtgga ggggggccct aacgtggtgt ccatcgtctt ggaatacctg caagaagagc
      601 ctcccatccc tgtggtgatt tgtgatggca gcggacgtgc ctcggacatc ctgtcctttg
      661 cgcacaagta ctgtgaagaa ggcggaataa taaatgagtc cctcagggag cagcttctag
      721 ttaccattca gaaaacattt aattataata aggcacaatc acatcagctg tttgcaatta
      781 taatggagtg catgaagaag aaagaactcg tcactgtgtt cagaatgggt tctgagggcc
      841 agcaggacat cgagatggca attttaactg ccctgctgaa aggaacaaac gtatctgctc
      901 cagatcagct gagcttggca ctggcttgga accgcgtgga catagcacga agccagatct
      961 ttgtctttgg gccccactgg ccgcccctgg gaagcctggc acccccgacg gacagcaaag
     1021 ccacggagaa ggagaagaag ccacccatgg ccaccaccaa gggaggaaga ggaaaaggga
     1081 aaggcaagaa gaaagggaaa gtgaaagagg aagtggagga agaaactgac ccccggaaga
     1141 tagagctgct gaactgggtg aatgctttgg agcaagcgat gctagatgct ttagtcttag
     1201 atcgtgtcga ctttgtgaag ctcctgattg aaaacggagt gaacatgcaa cactttctga
     1261 ccattccgag gctggaggag ctttataaca caagactggg tccaccaaac acacttcatc
     1321 tgctggtgag ggatgtgaaa aagagcaacc ttccgcctga ttaccacatc agcctcatag
     1381 acatcgggct cgtgctggag tacctcatgg gaggagccta ccgctgcaac tacactcgga
     1441 aaaactttcg gaccctttac aacaacttgt ttggaccaaa gaggcctaaa gctcttaaac
     1501 ttctgggaat ggaagatgat gagcctccag ctaaagggaa gaaaaagaaa aagaagaaaa
     1561 aggaggaaga gatcgacatt gatgtggacg accctgccgt gagtcggttc cagtatccct
     1621 tccacgagct gatggtgtgg gcagtgctga tgaaacgcca gaaaatggca gtgttcctct
     1681 ggcagcgagg ggaagagagc atggccaagg ccctggtggc ctgcaagctc tacaaggcca
     1741 tggcccacga gtcctccgag agtgatctgg tggatgacat ctcccaggac ttggataaca
     1801 attccaaaga cttcggccag cttgctttgg agttattaga ccagtcctat aagcatgacg
     1861 agcagatcgc tatgaaactc ctgacctacg agctgaaaaa ctggagcaac tcgacctgcc
     1921 tcaaactggc cgtggcagcc aaacaccggg acttcattgc tcacacctgc agccagatgc
     1981 tgctgaccga tatgtggatg ggaagactgc ggatgcggaa gaaccccggc ctgaaggtta
     2041 tcatggggat tcttctaccc cccaccatct tgtttttgga atttcgcaca tatgatgatt
     2101 tctcgtatca aacatccaag gaaaatgagg atggcaaaga aaaagaagag gaaaatacgg
     2161 atgcaaatgc agatgctggc tcaagaaagg taagtatcaa ggttacaaga caggtttaag
     2221 gagaccaata agctattatt cgtataggat actttatacg aagttattaa gttatggaga
     2281 ccggtgtggc acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact
     2341 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     2401 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg
     2461 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
     2521 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     2581 tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc
     2641 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     2701 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     2761 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     2821 atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg
     2881 atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa
     2941 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
     3001 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     3061 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     3121 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     3181 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     3241 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3301 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3361 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     3421 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     3481 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     3541 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     3601 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     3661 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     3721 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     3781 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     3841 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     3901 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     3961 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     4021 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     4081 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     4141 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     4201 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     4261 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     4321 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     4381 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4441 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4501 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     4561 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4621 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4681 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4741 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4801 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4861 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     4921 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     4981 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     5041 ctcacatgtc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc cgggcgtcgg
     5101 gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac
     5161 tccatcacta ggggttcctg cggccgcacg cgtgtcgact ctagaggatc tgcgatcgct
     5221 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag
     5281 gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg
     5341 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag
     5401 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagctg aagcttcgag
     5461 gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat ccacgccggt
     5521 tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc gccgtctagg
     5581 taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg agcctaccta
     5641 gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctac gtctttgttt
     5701 cgttttctgt tctgcgccgt tacagatcca agctgtgacc ggcgcctacc gtacggccac
     5761 catgtaccct tatgacgtac ctgactatgc tggatctatg ggtcagaaat cttggataga
     5821 gaaaaccttt tgcaaacggg aatgtatctt tgtaattcct agcatgaaag actctaacag
     5881 gtgttgctgt ggccagttca ccaaccagca tatcccccct ctgccaagtg caacacccag
     5941 caaaaatgaa gaggaaagca aacaggtgga gactcagcct gagaaatggt ctgttgccaa
     6001 gcacacccag agctacccaa cagattccta tggagttctt gaattccagg gtggcggata
     6061 ttccaataaa gccatgt
//
