LOCUS       sequence-489077-        6901 bp    DNA     circular SYN 09-OCT-2025
DEFINITION  5' AAVLINK plasmid for SETD1A expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6901)
  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  JOURNAL   Unpublished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 6901)
  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            "Unpublished"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'-SETD1A
FEATURES             Location/Qualifiers
     source          1..6901
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      723..1311
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1212..1231
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1373..1502
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        1559..1770
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             1783..2051
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             2088..2114
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     repeat_region   5536..5676
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5536..5665
                     /label=AAV2 ITR
     rep_origin      5751..6206
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5838..5857)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6048..6069
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6223..6242)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6342..6364
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6402..6420)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6488..6592
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6593..6661,6662..6901,1..552)
                     /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(6811..6830)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
       61 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
      121 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
      181 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
      241 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
      301 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
      361 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
      421 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
      481 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
      541 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
      601 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
      661 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
      721 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
      781 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
      841 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
      901 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
      961 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     1021 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     1081 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     1141 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     1201 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     1261 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     1321 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gtcctgcagg
     1381 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcgt cgggcgacct ttggtcgccc
     1441 ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc
     1501 ctgcggccgc acgcgtgtcg actctagagg atctgcgatc gctccggtgc ccgtcagtgg
     1561 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacg
     1621 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc
     1681 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt
     1741 tttcgcaacg ggtttgccgc cagaacacag ctgaagcttc gaggggctcg catctctcct
     1801 tcacgcgccc gccgccctac ctgaggccgc catccacgcc ggttgagtcg cgttctgccg
     1861 cctcccgcct gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt aaagctcagg
     1921 tcgagaccgg gcctttgtcc ggcgctccct tggagcctac ctagactcag ccggctctcc
     1981 acgctttgcc tgaccctgct tgctcaactc tacgtctttg tttcgttttc tgttctgcgc
     2041 cgttacagat ccaagctgtg accggcgcct accgtacggc caccatgtac ccttatgacg
     2101 tacctgacta tgctggatct gatcaggaag gtgggggaga tgggcagaag gccccgagct
     2161 tccagtggcg gaactacaag ctcatcgtgg atcctgcctt ggaccctgcc ctgcgcaggc
     2221 cttctcagaa ggtgtaccgc tatgatggag tccacttcag tgtcaacgac tcaaagtata
     2281 taccagtcga agacctccaa gacccccgtt gccatgtcag gtccaaaaac agagactttt
     2341 ccctcccagt ccctaagttt aagctggacg agttctatat tggacagatt ccactgaagg
     2401 aagtgacttt tgcaaggctg aatgacaacg tgcgggagac cttcctgaag gatatgtgcc
     2461 gtaagtacgg tgaggtggaa gaggtagaga tcctccttca cccccgtacg cgcaagcacc
     2521 tgggcctggc ccgtgtgctc ttcaccagca ctcggggcgc caaggaaacg gtcaaaaacc
     2581 tccaccttac ctccgtcatg ggcaacatca tccatgccca gcttgacatc aaaggacaac
     2641 aacgaatgaa atactatgaa ctaattgtca atggctccta cacccctcag actgtgccca
     2701 ctgggggcaa ggccctgagt gagaagttcc aaggctcggg tgcagccact gagacggccg
     2761 aatcccgccg ccgctcttcc tctgacacag ctgcctaccc agcaggcacc actgcggtgg
     2821 gcactcctgg caacggcacc ccctgctccc aggacacaag cttctccagc agccgacaag
     2881 ataccccatc ttcctttggc cagttcacac ctcagtcctc ccaaggaacc ccctacacgt
     2941 ctcggggcag caccccctac tctcaggact ctgcctactc cagcagcacc acttcaacct
     3001 ccttcaagcc ccggcggtca gagaacagct accaagatgc cttttcccgc cgccacttct
     3061 ctgcatcttc agcctccaca accgcctcca cggccatcgc cgccaccact gcagccactg
     3121 cctcatcctc cgcctcttcc tcctcattgt cctcgtcctc ctcgtcatcc tcttcctcct
     3181 cgtcctctca gtttcgtagt tctgatgcaa actacccagc gtattatgaa agctggaatc
     3241 gctaccagcg ccatacttcc tacccaccac gccgggccac acgggaggaa ccccctggag
     3301 ccccttttgc tgaaaataca gctgagcgct tcccaccttc ttacacctcc tacctgcccc
     3361 ccgagcccag ccggcccacc gaccaggact accggcctcc tgcctcagag gctccacccc
     3421 cggagcctcc agaacctggt ggaggcgggg gtggaggagg gcccagccct gagagagaag
     3481 aagttcggac ttccccccgc ccagcctccc ctgcccgctc tggctcccca gccccggaga
     3541 ccaccaatga gagtgtgccc ttcgcccagc acagcagcct ggattcccgc atcgagatgc
     3601 tgctgaagga gcagcgctcc aagttttcct tcttggcctc tgacacagag gaggaggaag
     3661 agaacagcag catggtcctt ggggccagag atacagggag tgaggtgcct tctgggtcag
     3721 ggcatgggcc ctgcacaccc cctccggccc cagctaattt tgaggatgtg gcacctacag
     3781 ggagcgggga gccaggggct acccgggagt ctcccaaggc aaatggacag aaccaggctt
     3841 ctccatgctc ttctggagac gacatggaga tctccgacga cgaccggggt ggctcacccc
     3901 ctccggcccc gacgccccct cagcagcctc cgccacctcc ccctcccccg ccgcctcctc
     3961 ctccctacct ggcgtccctt cctcttggtt atcctcccca ccaacctgcc tacctcctcc
     4021 cacccagacc tgatgggccg ccgccccctg agtacccccc acctcctcca ccacccccgc
     4081 acatctatga ctttgtgaac tccttggagc tcatggaccg acttggggct cagtggggag
     4141 ggatgcccat gtccttccag atgcagaccc agatgttaac tcggctccat cagctgcggc
     4201 agggcaaggg attgattgcc gcctcagctg gcccccccgg tggggccttt ggggaggcct
     4261 tcctcccgtt tccacccccg caggaggcag cctacggctt gccgtatgct ctatatgcac
     4321 aggggcagga gggcagaggg gcatactcac gggaggccta ccacctgccc atgccaatgg
     4381 cagccgagcc cctgccctcc tcctcagtct cgggagagga ggcccggctg ccacccaggg
     4441 aagaagcaga gctggcagag ggcaagaccc tcccgacagc aggcaccgtg ggccgtgtgc
     4501 tcgccatgct ggtccaggag atgaagagca tcatgcagcg agacctcaac cgcaagatgg
     4561 tggagaacgt ggccttcgga gcctttgacc agtggtggga gagcaaggag gagaaggcca
     4621 agccattcca gaacgcggcc aagcagcaag ccaaggagga ggataaagag aagacgaagc
     4681 tgaaggagcc tggcctgctg tccctcgtgg actgggccaa gagcgggggc actacgggca
     4741 tcgaggcttt cgcctttggg tcagggctga gaggggccct gcggctgcct tcattcaagg
     4801 taaagcggaa agagccatcg gaaatttccg aggccagtga ggaaaagagg cctcgtccct
     4861 ccactcctgc tgaggaagat gaagacgacc ctgaacaaga gaaggaggct ggagagccag
     4921 gacgtccggg gaccaagccc ccgaagcggg acgaagagcg aggcaagacc cagggcaagc
     4981 accgcaagtc ctttgctctg gacagcgaag gggaggaggc atcccaggag tcctcctcgg
     5041 agaaggatga ggaggatgac gaggaagatg aggaagatga agatcgagag gaagctgtgg
     5101 ataccacaaa gaaggagaca gaggtgtcgg atggcgagga cgaggaaagc gattcgtctt
     5161 ccaaatgttc tctgtatgct gactcagatg gcgaaaatga cagcacatca gactccgaga
     5221 gcagcagctc ttccagctcc tcatcctcct cctcctcctc gtcctcatcc tcctcgtcct
     5281 cttcatcctc tgagtcctcc tctgaagatg aagaggaaga ggagcggcca gcagcccttc
     5341 cctcagcctc cccgcccccc agagaagtcc cagtgcccac gccagcacct gtggaggtgc
     5401 cagtgccgga aaggtaagta tcaaggttac aagacaggtt taaggagacc aataagctat
     5461 tattcgtata ggatacttta tacgaagtta ttaagttatg gagaccggtg tggcacgtgc
     5521 ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     5581 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     5641 cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc
     5701 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
     5761 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     5821 cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     5881 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     5941 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     6001 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     6061 ccaaactgga acaacactca actctatctc gggctattct tttgatttat aagggatttt
     6121 gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     6181 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
     6241 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     6301 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     6361 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     6421 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     6481 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     6541 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     6601 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     6661 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     6721 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     6781 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     6841 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     6901 c
//
