LOCUS       sequence-489083-        6665 bp    DNA     circular SYN 09-OCT-2025
DEFINITION  5' AAVLINK plasmid for KIF14 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6665)
  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  JOURNAL   Unpublished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 6665)
  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            "Unpublished"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'-KIF14
FEATURES             Location/Qualifiers
     source          1..6665
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   452..592
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   452..581
                     /label=AAV2 ITR
     rep_origin      667..1122
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(754..773)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     964..985
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1139..1158)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1258..1280
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1318..1336)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1404..1508
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1509..1577,1578..2369)
                     /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(1727..1746)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2540..3128
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3029..3048
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   3190..3319
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        3376..3587
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             3600..3868
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             3905..3931
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6088..6099
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 ctggaagaga tactcctata agtgagggtc caaaagactt tgaatttgca aaaaatgagt
       61 tgctcatggc acagagatca caacttgaag cagaaataaa agaggctcag ttgaaggcaa
      121 aggaagaaat gatgcaagga atccagattg caaaagaaat ggctcagcaa gagctttctt
      181 ctcaaaaagc tgcatatgaa agcaaaataa aagcactgga agcagaactg agagaagagt
      241 ctcaaaggaa aaaaatgcag gaaataaata accagaaggc taatcacaaa attgaggaat
      301 tagaaaaggc aaagcagcat cttgaacagg taagtatcaa ggttacaaga caggtttaag
      361 gagaccaata agctattatt cgtataggat actttatacg aagttattaa gttatggaga
      421 ccggtgtggc acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact
      481 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
      541 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg
      601 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
      661 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
      721 tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc
      781 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
      841 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
      901 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
      961 atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg
     1021 atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa
     1081 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
     1141 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     1201 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     1261 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     1321 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     1381 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     1441 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     1501 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     1561 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     1621 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     1681 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     1741 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     1801 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     1861 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     1921 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     1981 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     2041 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     2101 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     2161 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     2221 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     2281 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     2341 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     2401 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     2461 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     2521 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     2581 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     2641 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     2701 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     2761 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     2821 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     2881 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     2941 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     3001 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     3061 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     3121 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     3181 ctcacatgtc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc cgggcgtcgg
     3241 gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac
     3301 tccatcacta ggggttcctg cggccgcacg cgtgtcgact ctagaggatc tgcgatcgct
     3361 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag
     3421 gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg
     3481 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag
     3541 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagctg aagcttcgag
     3601 gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat ccacgccggt
     3661 tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc gccgtctagg
     3721 taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg agcctaccta
     3781 gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctac gtctttgttt
     3841 cgttttctgt tctgcgccgt tacagatcca agctgtgacc ggcgcctacc gtacggccac
     3901 catgtaccct tatgacgtac ctgactatgc tggatcttca ttacacagta ctcataatag
     3961 aaataacagc ggtgatattc ttgatattcc ttcttcccaa aatagttcat cactgaatgc
     4021 cctcacccac agtagccgac ttaagctgca tttgaagtcg gatatgtcag aatgtgaaaa
     4081 tgatgatcca ttattgagat ctgcaggtaa agtcagagac ataaatagaa cttatgttat
     4141 ttctgccagt agaaaaacag cagacatgcc ccttacccct aatcctgtag gtagattggc
     4201 acttcagagg agaactacaa ggaacaaaga atcatctttg cttgttagtg agttggaaga
     4261 cacaactgaa aaaacagcag aaacacgtct tacattacaa cgtcgtgcta aaacagattc
     4321 tgcagaaaag tggaaaacag ctgaaataga ttctgtcaaa atgacactga atgtgggagg
     4381 tgaaacagaa aataatggtg tttctaagga aagtagaaca aatgtaagga ttgtaaataa
     4441 tgctaaaaac tcttttgttg cctcttctgt acctttagat gaagatccac aggtcattga
     4501 aatgatggct gataagaaat acaaagaaac attttctgcc cccagtagag caaatgaaaa
     4561 tgttgcactt aagtactcaa gtaatagacc acccattgct tccctgagtc agactgaagt
     4621 tgttagatca ggacacttga caacgaaacc tactcagagc aagttggata tcaaagtgtt
     4681 gggaacagga aacttgtatc atagaagtat tgggaaggaa attgcaaaaa cttcaaataa
     4741 atttgggagc ttagaaaaaa gaacacctac aaaatgtaca acagaacaca aactgacaac
     4801 aaagtgcagc ctgcctcagc ttaagagccc agctccatca atactgaaga atagaatgtc
     4861 taaccttcaa gttaaacaaa gaccaaaaag ttcctttctt gcaaataaac aggaaagatc
     4921 cgcagaaaat acaattcttc ccgaagaaga aactgtagtt cagaacacct ctgcaggaaa
     4981 agacccctta aaagtagaga atagtcaagt gacagtggca gtacgcgtaa gacctttcac
     5041 caagagagag aagattgaaa aagcatccca ggtagtcttc atgagtggga aagaaataac
     5101 tgtggaacac cctgacacga aacaagttta taattttatt tatgatgttt cattctggtc
     5161 ttttgatgaa tgtcatcctc actacgctag ccagacaact gtctatgaga agctagcagc
     5221 accactccta gaaagagcct tcgaaggctt caatacctgt ctttttgctt atggtcagac
     5281 tggctctgga aaatcatata cgatgatggg atttagtgaa gaaccaggaa taattccaag
     5341 attttgtgaa gatctttttt ctcaagtagc cagaaaacaa acccaagagg tcagctatca
     5401 cattgaaatg agcttctttg aagtatataa tgaaaaaatt cacgaccttc tggtttgtaa
     5461 agatgaaaat gggcagagaa agcaaccact gagagtgagg gaacatcctg tttatggacc
     5521 atatgttgaa gcactgtcaa tgaacattgt cagttcttac gctgatatcc agagttggct
     5581 agaattggga aataaacaaa gagctactgc tgctactggt atgaatgata aaagttcccg
     5641 atctcattca gttttcaccc tggtgatgac ccagaccaag acagaatttg tggaagggga
     5701 agaacacgat cacagaataa caagtcgaat taacctaata gatctggcag gcagtgagcg
     5761 ctgctctacg gctcacacta atggagatcg actaaaggaa ggtgtgagta ttaataagtc
     5821 cttgctaact ttgggaaaag ttatatctgc actttcggaa caagcaaacc aaaggagtgt
     5881 ttttattcct tatcgtgaat ctgttcttac atggctgtta aaagaaagtc tgggtggaaa
     5941 ttcaaaaact gcaatgattg ctacgattag tcccgctgcc agcaacatag aagaaacatt
     6001 aagcacactt agatatgcta accaagcccg tttaatagtc aacattgcta aagtaaatga
     6061 agatatgaac gctaagttaa ttagagaatt gaaggcagaa attgcaaagc taaaagctgc
     6121 tcagagaaac agtcggaata ttgaccctga acgatacagg ctctgtcggc aagaaataac
     6181 atccttaaga atgaaactgc atcaacagga gagagacatg gcagaaatgc aaagagtgtg
     6241 gaaagaaaag tttgaacaag ctgaaaaaag aaaacttcaa gaaacaaaag agttacagaa
     6301 agcaggaatt atgtttcaaa tggacaatca tttaccaaac cttgttaatc tgaatgaaga
     6361 tccacaacta tctgagatgc tgctatatat gataaaagaa ggaacaacta cagttggaaa
     6421 gtataaacca aactcaagcc atgatattca gttatctggg gtgctgattg ctgatgatca
     6481 ttgtactatc aaaaattttg gtgggacagt gagtattatc ccagttgggg aagcaaagac
     6541 atatgtaaat ggaaaacata ttttggaaat cacagtatta cgtcatggtg atcgagtgat
     6601 tcttggtgga gatcattatt ttagatttaa tcatccagta gaagtccaga aaggaaaaag
     6661 gccat
//
