LOCUS       sequence-491435-        7203 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5' AAVLINK plasmid for TCF20 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7203)
  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  JOURNAL   Unpublished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 7203)
  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            "Unpublished"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'-TCF20
FEATURES             Location/Qualifiers
     source          1..7203
                     /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        175..386
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             399..667
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             704..730
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     rep_origin      4681..5136
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4768..4787)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4978..4999
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5153..5172)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5272..5294
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5332..5350)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5418..5522
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5523..5591,5592..6383)
                     /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(5741..5760)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6554..7142
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7043..7062
                     /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 gcgtggatct gcgatcgctc cggtgcccgt cagtgggcag
      181 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaacgggtg
      241 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
      301 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
      361 gcaacgggtt tgccgccaga acacagctga agcttcgagg ggctcgcatc tctccttcac
      421 gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc
      481 ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga
      541 gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg ctctccacgc
      601 tttgcctgac cctgcttgct caactctacg tctttgtttc gttttctgtt ctgcgccgtt
      661 acagatccaa gctgtgaccg gcgcctaccg tacggccacc atgtaccctt atgacgtacc
      721 tgactatgct ggatctcagt cctttcggga gcaaagcagt taccacggaa accagcaaag
      781 ctacccacag gaggtacacg gctcatcccg gctagaagag ttcagccctc gtcaggccca
      841 gatgttccag aattttggag gtacaggtgg cagtagtggc agcagtggca gtggcagtgg
      901 tggtggacga cgaggagcag cagctgctgc ggcagcgatg gctagcgaga cctctggcca
      961 tcaaggttac cagggtttca ggaaagaggc tggagatttt tactacatgg caggcaacaa
     1021 agaccccgtg actacaggaa ccccacagcc tcctcagcga aggccttctg ggcctgtgca
     1081 gagctatgga cccccccagg ggagcagctt tggcaatcag tatgggagtg agggtcatgt
     1141 gggccagttt caagcacagc actctggcct tggcggtgtg tcacattatc agcaggatta
     1201 cactgggcct ttctctccag ggagtgctca gtaccaacag caggcttcca gccagcagca
     1261 gcagcagcaa gtccagcagt tgagacaaca gctttaccag tcccatcagc ccctgccaca
     1321 ggccactggc caaccagcat ccagctcatc ccatctacag ccaatgcagc ggccctcaac
     1381 tctgccatcc tctgctgctg gttaccagtt aagagtgggt cagtttggcc aacactatca
     1441 gtcttctgct tcctcctcct cctcctcctc cttcccttca ccacagcgtt ttagccagtc
     1501 tggacagagc tatgatggca gttacaatgt gaatgctgga tctcagtatg aaggacacaa
     1561 tgtgggttct aatgcacagg cttatggaac acaatccaat tacagctatc agcctcaatc
     1621 tatgaagaat tttgaacagg caaagattcc acaagggacc caacaggggc agcagcagca
     1681 gcaaccgcag caacaacaac acccttctca gcatgtgatg cagtatacta acgctgccac
     1741 caagctgccc ctgcaaagcc aagtggggca gtacaaccag cctgaggttc ctgtgaggtc
     1801 ccccatgcag tttcaccaga acttcagccc catttctaac ccttctccag ctgcctctgt
     1861 ggttcagtct ccaagctgta gttctacccc atctcctctc atgcagactg gggagaatct
     1921 ccagtgtggg caaggcagtg tgcctatggg ttccagaaac agaattttac agttaatgcc
     1981 tcaactcagt ccaaccccat caatgatgcc cagtcctaat tctcatgctg caggcttcaa
     2041 agggtttgga ctagaagggg taccagaaaa gcgactgaca gatcctgggt tgagtagttt
     2101 gagtgctctg agtactcaag tggccaatct tcctaacact gtccagcaca tgttactttc
     2161 tgatgccctg actcctcaga agaagacctc caagaggccc tcatcttcca agaaagcaga
     2221 tagctgcaca aattctgaag gctcctcaca acctgaagaa cagctgaagt cccctatggc
     2281 agagtcatta gatggaggct gctccagcag ttcagaggat caaggcgaga gagtgcggca
     2341 actaagtggc cagagcacca gctctgacac cacctacaag ggtggagcct ctgagaaagc
     2401 tggctcctca ccggcacaag gtgctcagaa tgaacccccc agactcaatg ctagtcctgc
     2461 cgcaagagaa gaggccacct caccaggcgc taaggacatg ccattgtcat ccgacgggaa
     2521 cccaaaggtt aatgagaaga ctgttggggt gattgtctcc cgggaagcca tgacaggtcg
     2581 ggtagaaaag cctggtggac aagataaagg ctcccaagag gatgatcctg cagccactca
     2641 aaggccacct agcaatggtg gggcaaagga aaccagtcat gcatcacttc cccagccaga
     2701 gcctccagga ggaggaggga gcaaaggaaa caagaatggc gataacaact ccaaccataa
     2761 tggagaagga aatggccaga gtggccactc tgcagcgggc cctggtttta cgagcagaac
     2821 tgagcctagc aaatctcctg gaagtctgcg ctatagttac aaagatagtt tcgggtcagc
     2881 cgtgccacga aatgtcagtg gctttcctca gtatcctaca gggcaagaaa agggagattt
     2941 cactggccat ggggaacgaa agggtagaaa tgaaaaattc ccaagcctcc tgcaggaagt
     3001 gcttcagggt taccaccacc accctgacag gagatattct aggagtactc aagagcatca
     3061 ggggatggct ggtagcctag aaggaaccac aaggcccaat gtcttggtta gtcaaaccaa
     3121 tgaattagct agcaggggcc ttctgaacaa aagcattggg tctctattag aaaatcccca
     3181 ctggggcccc tgggaaagga aatcaagcag cacagctcct gaaatgaaac agatcaattt
     3241 gactgactat ccaattccca gaaagtttga aatagagcct cagtcatcag cacatgagcc
     3301 tgggggttcc ctctctgaaa gaagatcagt gatctgtgat atttctccac taagacagat
     3361 tgtcagggac ccaggggctc actcactggg acacatgagt gccgacacca gaattgggag
     3421 gaatgaccgt ctcaatccaa ctttaagtca gtcggtcatt cttcctggtg gtttggtgtc
     3481 catggaaacc aagctgaaat cccagagcgg gcagataaaa gaggaagact ttgaacagtc
     3541 taaatctcaa gctagtttca acaacaagaa atctggagac cactgccatc ctcctagcat
     3601 caagcatgag tcttaccgcg gcaatgccag ccctggagca gcaacccatg attccctttc
     3661 agactatggc ccgcaagaca gcagacccac gccaatgcgg cgggtccctg gcagagttgg
     3721 tggtcgggag ggcatgaggg gtcggtcccc ttctcaatat catgactttg cagaaaaatt
     3781 gaaaatgtct cctgggcgga gcagaggccc agggggagac cctcatcaca tgaatccaca
     3841 catgaccttt tcagagaggg ctaaccggag ttctttacac actccctttt ctcccaactc
     3901 agaaaccctg gcctctgctt atcatgcaaa tactcgggct catgcttatg gggaccctaa
     3961 cgcaggtttg aattctcagc tgcattataa gagacagatg taccaacagc aaccagagga
     4021 gtataaagac tggagcagcg gttctgctca gggagtaatt gctgcagcac agcacaggca
     4081 ggaggggcca cggaagagtc caaggcagca gcagtttctt gacagagtac ggagccctct
     4141 gaaaaatgac aaagatggta tgatgtatgg cccaccagtg gggacttacc atgaccccag
     4201 tgcccaggag gctgggcgct gcctaatgtc tagtgatggt ctgcctaaca agggcatgga
     4261 attaaagcat ggctcccaga agttacaaga atcctgttgg gatctttctc ggcaaacttc
     4321 tccagccaaa agcagcggtc ctccaggaat gtccagtcaa aaaaggtaag tatcaaggtt
     4381 acaagacagg tttaaggaga ccaataagct attattcgta taggatactt tatacgaagt
     4441 tattaagtta tggagaccgg tgtggcacgt gcggaccgag cggccgcagg aacccctagt
     4501 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     4561 ggtcgcccgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     4621 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     4681 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4741 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4801 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4861 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     4921 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4981 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     5041 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     5101 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     5161 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     5221 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     5281 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     5341 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     5401 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     5461 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     5521 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     5581 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     5641 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     5701 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     5761 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5821 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5881 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5941 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     6001 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     6061 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     6121 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     6181 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     6241 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     6301 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     6361 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     6421 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     6481 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     6541 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     6601 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6661 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6721 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6781 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6841 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6901 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6961 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     7021 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     7081 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     7141 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     7201 tgt
//
