LOCUS       sequence-491887-        6835 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5' AAVLINK plasmid for CLTCL1 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6835)
  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  JOURNAL   Unpublished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 6835)
  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            "Unpublished"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-HA-5'-CLTCL1
FEATURES             Location/Qualifiers
     source          1..6835
                     /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"
     rep_origin      4313..4768
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4400..4419)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4610..4631
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4785..4804)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4904..4926
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4964..4982)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5050..5154
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5155..5223,5224..6015)
                     /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(5373..5392)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6186..6774
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6675..6694
                     /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 ctggatctat ggcgcagatc ctccctgttc gctttcagga
      781 gcacttccag ctccaaaacc ttggaattaa tccagctaac attggattca gcacactgac
      841 catggaatct gacaagttca tatgtatccg agagaaagtt ggtgagcagg cacaggtcac
      901 gatcattgac atgagtgacc caatggctcc gatccgacgg cctatctctg cagagagtgc
      961 catcatgaat ccagcctcta aggtgatagc tctgaaagct gggaagacac ttcagatctt
     1021 taatattgag atgaagagta aaatgaaggc tcatactatg gcagaagaag tgattttctg
     1081 gaaatgggtt tctgtgaaca ctgttgcctt ggtgaccgag accgcggtct accactggag
     1141 catggaaggt gactcccagc ccatgaagat gtttgataga cataccagtc tggtgggctg
     1201 ccaggtgatt cactaccgga ctgatgagta ccagaagtgg ctgctgctcg taggcatctc
     1261 ggctcagcaa aaccgtgtgg ttggagcaat gcagctctac tctgtggata ggaaggtttc
     1321 acaacccata gaaggccatg ctgcggcttt tgcagagttc aagatggagg ggaatgccaa
     1381 gcctgccacc cttttctgct ttgctgtacg taatcccaca ggaggcaagt tgcacatcat
     1441 tgaagttgga cagcctgcag cgggaaacca accttttgta aagaaagcag tagatgtgtt
     1501 ttttcctcca gaggcacaga atgattttcc agtggctatg cagattggag ctaaacatgg
     1561 tgttatttac ttgatcacaa agtatggcta tcttcatctg tacgacctag agtctggcgt
     1621 gtgcatctgc atgaaccgta ttagtgctga cacaatattt gtcactgctc cacacaaacc
     1681 aacctctgga attattggtg tcaacaaaaa gggacaggta ctgtcagttt gtgttgagga
     1741 agataacatt gtgaattatg caaccaacgt gcttcagaat ccagaccttg gtctgcgttt
     1801 ggccgttcgt agtaacctgg ctggggcaga gaagttgttt gtgagaaaat tcaataccct
     1861 ctttgcacag ggcagctatg ctgaagccgc caaagttgca gcgtctgcac caaagggaat
     1921 cctgcgtacc agagagacgg tccagaaatt ccagagtata cccgctcagt ctggccaggc
     1981 ttctccattg ctgcagtact tcggaatcct gctcgaccag ggtcagctca ataaacttga
     2041 atccttagaa ctttgccatc tggttcttca gcaggggcgt aagcaactcc tagagaagtg
     2101 gctgaaagaa gataagctgg agtgctcaga ggagctcgga gacttggtca aaaccactga
     2161 ccccatgctc gctctgagtg tgtaccttcg ggcaaatgtg ccaagcaaag tgatccagtg
     2221 ttttgcagaa acaggccaat tccagaaaat tgtgctctat gccaaaaagg ttgggtacac
     2281 cccagactgg atctttctgc tgaggggtgt aatgaagatc agtccggaac agggcctgca
     2341 gttttctcga atgctagtgc aggacgagga gccgctggcc aacattagcc agattgtgga
     2401 cattttcatg gaaaacagtt taattcagca gtgtacttcc ttcttattgg atgccttgaa
     2461 gaataatcgc ccagctgagg gactcctgca gacatggctg ttggagatga accttgttca
     2521 tgcaccccag gttgcagatg ccatccttgg aaataaaatg tttactcatt acgaccgggc
     2581 ccacattgcc cagctctgtg agaaggcagg cctcctgcag caagcactgg agcactacac
     2641 cgacctctat gacatcaaga gggctgtggt ccacactcac ctcctcaatc ccgagtggct
     2701 tgtcaatttc tttggctcct tatcggtgga ggattctgtg gagtgtctgc atgccatgct
     2761 gtctgctaac atcagacaga accttcagct gtgtgtgcag gtggcctcta agtaccacga
     2821 gcagctgggc acgcaggccc tggtggagct ctttgaatcc ttcaagagtt acaaaggcct
     2881 cttctacttc ctgggctcaa tcgtgaactt cagccaagac ccagatgtgc atctgaaata
     2941 cattcaggct gcctgtaaga cagggcagat caaggaggtg gagaggatat gccgagagag
     3001 cagctgctac aacccagagc gtgtgaagaa cttcctgaag gaggccaagc tcacagacca
     3061 gcttcccctc atcatcgtgt gtgatcgttt tggctttgtc catgaccttg tcctatattt
     3121 ataccgcaac aacctgcaga ggtacattga gatctacgtg cagaaggtca accctagccg
     3181 gaccccagct gtgattggag ggctgcttga tgtggattgt tctgaggaag tgattaaaca
     3241 cttaatcatg gcagtgagag gacagttctc tactgatgag ttggtggctg aagtagaaaa
     3301 aagaaatagg ctcaagctgc tgcttccctg gctggagtcc cagattcagg aaggctgtga
     3361 ggagcctgcc actcacaatg cactggctaa aatctacatc gacagcaaca acagccccga
     3421 gtgcttcctg agagagaatg cctactatga cagcagcgtg gtgggccgct actgtgagaa
     3481 gcgagacccc catctggcct gtgttgccta tgagcggggg cagtgtgacc ttgagctcat
     3541 caaggtgtgc aatgagaatt ctctgttcaa aagcgaggcc cgctacctgg tatgcagaaa
     3601 ggatccggag ctctgggctc acgtccttga ggagaccaac ccatccagga gacagctaat
     3661 tgaccaggtg gtacagacag cattgtcaga aacacgggat cctgaagaga tttcggtcac
     3721 tgtcaaagcc tttatgacag ccgacctgcc taatgaactg attgaactgc tggagaagat
     3781 agttctggat aactctgtct tcagcgagca caggaatcta cagaatctgt tgatcctgac
     3841 tgccatcaag gcagaccgca cacgggtcat ggagtacatc agccgcctgg acaactatga
     3901 cgcactggac atcgcgagca tcgctgtcag cagcgcactg tatgaggagg ccttcaccgt
     3961 tttccacaag tttgatatga atgcctcagc aatccaggta agtatcaagg ttacaagaca
     4021 ggtttaagga gaccaataag ctattattcg tataggatac tttatacgaa gttattaagt
     4081 tatggagacc ggtgtggcac gtgcggaccg agcggccgca ggaaccccta gtgatggagt
     4141 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     4201 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
     4261 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     4321 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4381 gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4441 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     4501 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     4561 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4621 ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt
     4681 ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4741 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     4801 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     4861 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     4921 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     4981 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     5041 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     5101 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     5161 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     5221 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     5281 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5341 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     5401 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5461 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5521 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5581 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5641 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5701 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5761 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5821 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     5881 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5941 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     6001 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     6061 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6121 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6181 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6241 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6301 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     6361 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6421 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6481 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6541 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6601 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6661 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6721 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6781 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgt
//
