LOCUS       sequence-491924-        7076 bp    DNA     circular SYN 05-NOV-2025
DEFINITION  5' AAVLINK plasmid for KIF1A expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7076)
  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  JOURNAL   Unpublished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 7076)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 5, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pAAVLINK-EFS-HA-5'-KIF1A
FEATURES             Location/Qualifiers
     source          1..7076
                     /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        145..356
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             371..397
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     repeat_region   4339..4479
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4339..4468
                     /label=AAV2 ITR
     rep_origin      4554..5009
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4641..4660)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4851..4872
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5026..5045)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5145..5167
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5205..5223)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5291..5395
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5396..5464,5465..6256)
                     /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(5614..5633)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6427..7015
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6916..6935
                     /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 gcgtgggcag agcgcacatc gcccacagtc cccgagaagt
      181 tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg gtaaactggg
      241 aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa
      301 gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacagcgta
      361 cgccaccatg tacccttatg acgtacctga ctatgctgga tctgccgggg cttcggtgaa
      421 ggtggcggtg cgggtccgcc ccttcaattc ccgggaaatg agccgtgact ccaagtgcat
      481 cattcagatg tctggaagca ccaccaccat tgttaacccc aaacagccca aggagacgcc
      541 caaaagcttc agctttgact actcctactg gtcgcacacc tcacctgagg acatcaacta
      601 cgcgtcgcag aagcaggtgt accgggacat cggcgaggag atgctgcagc atgcctttga
      661 gggatacaac gtgtgcatct tcgcctatgg gcagacgggt gccggcaagt cctacaccat
      721 gatgggcaag caggagaagg accagcaggg catcatccca cagctctgcg aggacctctt
      781 ctctcggatc aacgacacga ccaacgacaa catgtcctac tccgtggagg tcagctacat
      841 ggagatttac tgtgagcgcg tccgtgacct cctgaacccc aagaacaagg gcaaccttcg
      901 cgtgagggag cacccactgc tggggcccta cgtggaggac ctctccaagc tggctgtcac
      961 ctcctacaat gacatccagg acctcatgga ctcagggaac aaggccagga ccgtggcggc
     1021 caccaacatg aatgagacca gcagtcgctc ccacgccgtc ttcaacatca tcttcaccca
     1081 gaagcgccat gacgcagaga ccaatatcac cacggagaag gtgagcaaaa tcagcctggt
     1141 ggacctggct gggagcgagc gggctgactc cacgggagcc aagggcacgc gcctcaagga
     1201 gggggccaac atcaacaagt cgctgaccac cctgggcaag gtcatctccg ccctggctga
     1261 aatggactcc ggacccaaca agaacaagaa aaagaagaag acagatttca ttccgtaccg
     1321 agattccgtg ttgacctggc tcctccggga aaacctgggc ggtaactcaa ggacagctat
     1381 ggtggcagcc ttgagtcctg cagacatcaa ctacgatgag acccttagca cgctgaggta
     1441 tgctgaccgg gccaagcaga tccgctgcaa tgctgtcatc aatgaggacc ccaacaacaa
     1501 gctgatccgc gagctgaagg atgaggtgac ccggctgcgg gaccttctgt acgcccaggg
     1561 tcttggcgac atcactgaca ccaacactgt gcctggagga cccaaattga ccaatgccct
     1621 ggtgggtatg agcccctcat cctcgctctc agccctgtcc agccgcgcgg cctccgtgtc
     1681 cagcctccac gagcgcatct tgtttgcccc gggcagcgag gaggccattg aaagactgaa
     1741 ggaaacagag aagatcatag ctgagctcaa tgagacctgg gaggagaagc tgcggcggac
     1801 agaagccatc cggatggaga gggaagccct gctggccgag atgggtgtgg ccatgaggga
     1861 ggatggcggc accttgggcg tattctctcc caaaaagaca ccacacctcg tcaacctgaa
     1921 cgaggacccg ctgatgtctg agtgcctgct ctactacatc aaggatggga tcaccagagt
     1981 gggcagggag gatggcgaga ggcggcagga cattgttctg agtgggcact tcatcaagga
     2041 ggagcactgc gtcttccgga gcgactccag gggaggcagc gaagctgtgg tgaccttgga
     2101 gccctgtgag ggggcagaca cctacgtcaa tggcaagaaa gtcacagagc ccagcatcct
     2161 gcgttcagga aaccgcatca tcatgggtaa gagtcatgtg ttccggttca accaccccga
     2221 gcaggcccgg caggagcgtg agcgcacgcc ttgtgcggag acgccagctg agcctgtgga
     2281 ctgggccttc gcccagcgtg agctgctgga gaagcagggc atcgacatga agcaggagat
     2341 ggagcagagg ctccaggaac tggaggacca gtaccgccgc gagcgggagg aggccaccta
     2401 cctgctggag cagcagcggc tggactatga gagcaagctg gaggctctgc agaagcagat
     2461 ggactccagg tactacccgg aggtgaacga ggaggaggag gagcccgagg atgaagtcca
     2521 gtggacagag cgggagtgtg agctggcgct ctgggccttc cggaagtgga agtggtacca
     2581 gttcacgtct ctgcgggacc tgctgtgggg caacgccatc ttcctcaagg aggccaatgc
     2641 catcagcgtg gagctgaaaa agaaggtaca attccagttt gtcctcctga cggacacact
     2701 ctactcccct ctgccacccg acctgctgcc cccagaggcc gccaaagacc gagagacgcg
     2761 gcccttcccc cgcaccattg tggccgtgga ggtccaggac cagaagaacg gggccaccca
     2821 ctactggacg ctggagaagc tcaggcagcg tctggacctg atgcgggaga tgtacgaccg
     2881 cgctgcagag gtgccctcca gtgtcatcga ggactgtgac aacgtggtga ccggcggaga
     2941 ccccttctat gaccgcttcc cctggttccg gctggtgggc agttcagcca tctctggctg
     3001 caacagctac cctcttctca acacatgcat gagcgagcgc atggctgctc tcaccccctc
     3061 ccccaccttc tcgagccccg actccgacgc caccgagcct gccgaggagc agagcgtggg
     3121 ggaggaggag gaggaggagg aggaggagga ggatgaggag gaggaggacc tggaggacga
     3181 cgtctttccg gagcacgcgc tgtgcgacgg ccgggacccg ttttacgacc ggccccccct
     3241 gttcagttta gtaggaaggg ccttcgtgta cctgagcaac ctgctgtacc ccgttcccct
     3301 ggtacaccgt gtggcaatcg tcagcgagaa gggcgaggtg aagggcttcc tccgcgtggc
     3361 cgtccaggcc atctcagccg atgaagaggc ccctgattat ggctctggcg tccgccagtc
     3421 gggaactgct aaaatctcct ttgatgacca gcattttgaa aagttccagt ccgagtcttg
     3481 ccccgtggtg gggatgtccc gctcgggaac ctcccaggaa gagcttcgca tcgtggaggg
     3541 ccagggccag ggtgcagacg tggggccctc agccgatgaa gtcaacaaca acacctgttc
     3601 agcagtgccc ccagaaggcc tcctcctaga cagctctgag aaagccgccc tggatgggcc
     3661 cctggatgct gccctggacc acctccgcct gggcaacacc ttcaccttcc gtgtgacagt
     3721 cctgcaggcg tccagcatct ctgccgaata tgccgacatc ttctgccagt tcaacttcat
     3781 ccaccgccac gacgaggcct tctccacaga gcccctgaag aacacaggca gaggcccccc
     3841 acttggcttc taccacgtcc agaacatcgc agtggaggtg accaagtcct tcattgagta
     3901 catcaagagc cagcccattg ttttcgaggt ctttggccac taccagcagc acccgttccc
     3961 gcccctctgc aaggacgtgc tcagccccct gaggccctcg cgccgccact tccctcgggt
     4021 catgccactg tccaagccag tgcccgccac caagctcagc acactgacgc ggccctgtcc
     4081 gggaccctgc cactgcaagt acgacctgct ggtctacttc gagatctgtg agctggaggc
     4141 caacggcgat tacatcccgg ccgtggtgga ccaccgtggg ggcatgccat gcatggggac
     4201 cttcctcctc caccaggtaa gtatcaaggt tacaagacag gtttaaggag accaataagc
     4261 tattattcgt ataggatact ttatacgaag ttattaagtt atggagaccg gtgtggcacg
     4321 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     4381 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     4441 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc
     4501 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     4561 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4621 tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4681 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4741 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     4801 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4861 gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat
     4921 tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4981 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     5041 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     5101 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     5161 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     5221 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     5281 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     5341 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     5401 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     5461 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     5521 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5581 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     5641 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5701 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5761 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5821 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5881 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5941 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     6001 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     6061 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     6121 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     6181 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     6241 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     6301 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6361 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6421 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6481 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6541 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     6601 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6661 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6721 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6781 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6841 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6901 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6961 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     7021 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
