LOCUS       sequence-480231-        7043 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5' AAVLINK plasmid for PREX1 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7043)
  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  JOURNAL   Unpublished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 7043)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 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            "Unpublished"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' PREX1
FEATURES             Location/Qualifiers
     source          1..7043
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        187..398
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             411..679
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             716..742
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     repeat_region   4306..4446
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4306..4435
                     /label=AAV2 ITR
     rep_origin      4521..4976
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4608..4627)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4818..4839
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4993..5012)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5112..5134
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5172..5190)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5258..5362
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5363..5431,5432..6223)
                     /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(5581..5600)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6394..6982
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6883..6902
                     /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 gcgtgtcgac tctagaggat ctgcgatcgc tccggtgccc
      181 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
      241 attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
      301 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
      361 acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca
      421 tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg
      481 ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa
      541 agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc
      601 ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg
      661 ttctgcgccg ttacagatcc aagctgtgac cggcgcctac cgtacggcca ccatgtaccc
      721 ttatgacgta cctgactatg ctggatctga ggcgcccagc ggcagcgagc ccggcggcga
      781 cggggccggg gactgcgccc acccggaccc ccgggcccct ggcgccgcgg cgcccagctc
      841 cggccccggc ccgtgcgcgg ccgcccggga gtccgagcgc cagctgcgcc tccgcctctg
      901 cgtcctcaac gagatcttgg gcaccgagag ggactacgtg ggcaccttgc gcttcttgca
      961 gtcggcattc ctgcatcgca tccggcagaa cgtggccgac tcagtggaga agggcctcac
     1021 ggaggagaat gtcaaggtcc tgttctcgaa catcgaagac atcctggaag ttcataagga
     1081 tttcttggcc gccttggagt attgtttaca cccggagccg cagtctcagc atgaacttgg
     1141 gaatgttttc ttaaaattca aggacaagtt ctgcgtgtac gaggagtatt gcagcaacca
     1201 tgagaaagcc ctgaggctgc tggtggagct gaacaagatc cctaccgtgc gcgccttcct
     1261 tttgagctgc atgcttctgg gaggccggaa gaccacggac atccctttgg aaggctacct
     1321 gttgtctccg atccagagga tctgcaagta cccgctcctc cttaaggagc tggccaagag
     1381 gactcccggc aagcacccag accaccccgc ggtccagagt gccctgcagg ccatgaagac
     1441 cgtttgctcc aacatcaatg agaccaagcg gcagatggag aagctggaag ccctggagca
     1501 gctgcagtcc cacatcgaag gctgggaggg ttccaacctc acagacatct gcactcagct
     1561 cctcctgcaa gggactttgt taaagatctc tgcgggcaac atccaggaaa gggccttctt
     1621 cctcttcgac aaccttctcg tctactgcaa gcggaaatcc agggtcaccg ggagcaagaa
     1681 gtccaccaag aggaccaaat ccatcaacgg ctccctctac atcttcaggg gtcgaatcaa
     1741 cactgaagtc atggaggtgg agaatgtgga agatgggaca gcggattacc atagcaacgg
     1801 ctataccgtc accaacggct ggaagatcca caacacggcc aagaataagt ggtttgtctg
     1861 catggccaag acggcagagg agaagcagaa gtggctggat gccatcatcc gcgagcggga
     1921 gcagcgcgag agcctgaagc tgggcatgga gcgtgatgcc tacgtcatga ttgcggagaa
     1981 gggggagaag ctgtaccaca tgatgatgaa caagaaggtg aacctcatca aggaccgccg
     2041 gagaaagctg agcactgtcc ccaagtgctt tcttggcaat gagttcgttg cctggctcct
     2101 agaaattggt gaaatcagca agacggaaga aggagtcaac ttgggccaag ccctgttgga
     2161 gaatggcatc atccaccatg tttccgacaa gcaccagttc aagaatgagc aggtgatgta
     2221 tcgcttccgc tacgacgatg gcacctacaa ggcccgaagt gagctggagg acatcatgtc
     2281 caagggtgtg aggctttact gccgtcttca cagcctctac accccggtga tcaaagaccg
     2341 tgattaccac ctgaagacct acaagtcagt gcttcccggg agcaagctgg tggactggct
     2401 gctggctcag ggagactgcc agactcggga ggaggcagtg gcgctcggcg tgggtctgtg
     2461 caacaatggc ttcatgcacc acgtgctgga gaagagcgag ttcagggatg agtcccagta
     2521 cttccgcttt catgctgacg aggagatgga ggggaccagc agcaagaaca aacagcttcg
     2581 caacgacttc aagctggtgg agaacattct ggccaagcgc ctgctgatcc tgccccagga
     2641 ggaggactat ggctttgaca tcgaggagaa gaacaaggct gtggtggtga agtccgtcca
     2701 gaggggctcg ctggctgagg tggctggcct gcaggtgggg aggaagatct actccatcaa
     2761 tgaggacctg gtgttcctgc ggccgttttc agaggtggag tccatcctca accagtcctt
     2821 ctgctcccgc cgccctctgc gcctcctggt ggccacgaag gccaaagaga tcatcaaaat
     2881 ccccgaccag ccggacacac tgtgcttcca gattcgtgga gctgccccac cgtacgtcta
     2941 tgctgtgggg agaggctctg aggccatggc tgcagggctc tgtgctggtc agtgcattct
     3001 gaaggtcaat ggcagcaacg tgatgaacga tggtgcccct gaggtcctgg agcacttcca
     3061 ggcattccgg agtcggcgcg aagaggccct gggcctgtac cagtggatct accacaccca
     3121 tgaggatgcc caggaagcac gagccagtca ggaggcctcc actgaggacc ccagtggcga
     3181 gcaggcccag gaggaagacc aggctgattc agccttccca ctgctgtccc tgggtccccg
     3241 gctgagcctg tgtgaggaca gccccatggt caccctgact gtggacaacg tgcacctgga
     3301 acacggcgtg gtgtatgagt atgtgagcac ggcaggcgtc aggtgccatg tgctggagaa
     3361 gatcgtggag ccccgcggct gcttcggcct caccgccaag atcctcgagg cctttgctgc
     3421 caatgacagc gtcttcgtgg agaactgcag gcggctcatg gccctgagca gcgccatcgt
     3481 gaccatgccc cactttgagt tccgcaacat ctgtgacacc aagctggaga gcattggcca
     3541 gaggattgcc tgctaccagg agtttgcagc ccaactgaag agcagggtca gcccaccctt
     3601 caaacaagcc cccctggagc cccacccgct gtgtggcctg gacttctgcc ccaccaattg
     3661 ccacatcaac ctcatggaag tgtcctaccc caagaccacc ccctcagtgg gcaggtcctt
     3721 cagcatccgc tttggacgca aaccctccct catcggcctt gacccggagc aaggccacct
     3781 gaaccccatg tcgtacaccc agcactgcat caccaccatg gctgctccct cctggaagtg
     3841 cttgcctgct gcagagggtg atccccaagg ccagggtctc catgatggca gcttcgggcc
     3901 agccagtggg acccttggtc aggaagaccg gggcctcagc ttcctactca agcaggagga
     3961 ccgtgagatc caggatgcct acctgcagct cttcaccaag ctggatgtgg ccctgaagga
     4021 gatgaagcaa tatgtcaccc agatcaacag gctgctgtcc accatcacag agcccacctc
     4081 gggtgggtcc tgcgacgcat ccttggctga ggaggcctcc tccctgcccc tggtcagtga
     4141 agagagcgag atggacagga gtgaccatgg gggcatcaag aaggtaagta tcaaggttac
     4201 aagacaggtt taaggagacc aataagctat tattcgtata ggatacttta tacgaagtta
     4261 ttaagttatg gagaccggtg tggcacgtgc ggaccgagcg gccgcaggaa cccctagtga
     4321 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     4381 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     4441 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     4501 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4561 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
     4621 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4681 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     4741 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4801 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
     4861 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
     4921 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     4981 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     5041 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     5101 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     5161 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     5221 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     5281 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     5341 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     5401 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     5461 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     5521 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     5581 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     5641 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     5701 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     5761 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     5821 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     5881 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     5941 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     6001 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     6061 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     6121 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     6181 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     6241 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     6301 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     6361 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6421 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6481 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6541 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6601 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6661 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6721 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6781 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6841 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6901 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6961 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     7021 ttgctggcct tttgctcaca tgt
//
