LOCUS       sequence-479500-        6418 bp    DNA     circular SYN 23-JUL-2025
DEFINITION  3' AAVLINK plasmid for ERBIN expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6418)
  AUTHORS   Zhonghua Lu Lab
  TITLE     Zhonghua Lu AAV Plasmids 2025
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6418)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 23, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pAAVLINKv1-pCALM1-Cre-3' ERBIN
FEATURES             Location/Qualifiers
     source          1..6418
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             277..1308
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     primer_bind     1464..1483
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             complement(2453..2464)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             3138..3203
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3268..3288)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   3681..3821
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3681..3810
                     /label=AAV2 ITR
     rep_origin      3896..4351
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3983..4002)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4193..4214
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4368..4387)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4487..4509
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4547..4565)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4633..4737
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4738..4806,4807..5598)
                     /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(4956..4975)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5769..6357
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6258..6277
                     /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 gcgtcgctct gcgcgaggcg gctccgccgc ggcggaggga
      181 tacggcgcac catatatata tcgcggggcg cagactcgcg ctccggcagt ggtgctggga
      241 gtgtcgtgga cgccgtgccg ttactcgtac gccaccatgt ccaatttact gaccgtacac
      301 caaaatttgc ctgcattacc ggtcgatgca acgagtgatg aggttcgcaa gaacctgatg
      361 gacatgttca gggatcgcca ggcgttttct gagcatacct ggaaaatgct tctgtccgtt
      421 tgccggtcgt gggcggcatg gtgcaagttg aataaccgga aatggtttcc cgcagaacct
      481 gaagatgttc gcgattatct tctatatctt caggcgcgcg gtctggcagt aaaaaccatc
      541 cagcaacatt tgggccagct aaacatgctt catcgtcggt ccgggctgcc acgaccaagt
      601 gacagcaatg ctgtttcact ggttatgcgg cggatccgaa aagaaaacgt tgatgccggt
      661 gaacgtgcaa aacaggctct agcgttcgaa cgcactgatt tcgaccaggt tcgttcactc
      721 atggaaaata gcgatcgctg ccaggatata cgtaatctgg catttctggg gattgcttat
      781 aacaccctgt tacgtatagc cgaaattgcc aggatcaggg ttaaagatat ctcacgtact
      841 gacggtggga gaatgttaat ccatattggc agaacgaaaa cgctggttag caccgcaggt
      901 gtagagaagg cacttagcct gggggtaact aaactggtcg agcgatggat ttccgtctct
      961 ggtgtagctg atgatccgaa taactacctg ttttgccggg tcagaaaaaa tggtgttgcc
     1021 gcgccatctg ccaccagcca gctatcaact cgcgccctgg aagggatttt tgaagcaact
     1081 catcgattga tttacggcgc taaggatgac tctggtcaga gatacctggc ctggtctgga
     1141 cacagtgccc gtgtcggagc cgcgcgagat atggcccgcg ctggagtttc aataccggag
     1201 atcatgcaag ctggtggctg gaccaatgta aatattgtaa tgaactatat ccgtaacctg
     1261 gatagtgaaa caggggcaat ggtgcgcctg ctggaagatg gcgattaggt aagtatcaag
     1321 gttacaagac aggtttaagg agaccaataa ctagtataac ttcgtatagg atactttata
     1381 gcaatttatg aaactgggct tgtcgagaca gagaagactc ttgcgtttct gataggcacc
     1441 tattggtctt actgacatcc actttgcctt tctctccaca ggtaaatgga cattctgagg
     1501 aaacttccca gtctcctaat aggactgaac cacatgacag tgattgttct gttgacttag
     1561 gtatttccaa aagcactgaa gatctctccc ctcagaaaag tggtccagtt ggatctgttg
     1621 tgaaatctca tagcataact aatatggaga ttggagggct aaaaatctat gatattctta
     1681 gtgataatgg acctcagcag ccaagtacaa ccgttaaaat cacatctgct gttgatggaa
     1741 aaaatatagt caggagcaag tctgccacac tgttgtatga tcaaccattg caggtattta
     1801 ctggttcttc ctcatcttct gatttaatat caggaacaaa ggcaattttc aagtttgatt
     1861 caaatcataa tcccgaagag ccaaatataa taagaggccc cacaagtggc ccacaatctg
     1921 cacctcaaat atatggtcct ccacagtata atatccaata cagtagcagt gctgcagtca
     1981 aagacacttt gtggcactcc aaacaaaatc cccaaataga ccatgccagt tttcctcctc
     2041 agctccttcc tagatcagag agcacagaaa atcaaagtta tgctaaacat tctgccaata
     2101 tgaatttctc taatcataac aatgttcgag ctaatactgc ataccattta catcagagac
     2161 ttggcccagc aagacatggg gaaatgtggg ccatctcacc aaacgaccga cttattcctg
     2221 cagtaactcg aagtacaatc cagcgacaaa gtagtgtgtc ctccacagcc tctgtaaatc
     2281 ttggtgatcc aggctctaca aggcgggctc agattcctga aggagattat ttatcataca
     2341 gagagttcca ctcagcggga agaactcctc caatgatgcc aggatcacag agaccccttt
     2401 ctgcacgaac atacagcata gatggtccaa atgcatcaag acctcagagt gctcgaccct
     2461 ctattaatga aataccagag agaactatgt cagttagtga tttcaattat tcacggacta
     2521 gtccttcaaa aagaccaaat gcaagggttg gttctgagca ttctttatta gatcctccag
     2581 gaaaaagtaa agttcctcgt gactggagag aacaagtact tcgacatatt gaagccaaaa
     2641 agttagaaaa gatgcctttg agtaatggac agatgggcca gcctctcagg cctcaggcaa
     2701 attatagtca aatacatcac ccccctcagg catctgtggc aaggcatccc tctagagaac
     2761 aactaattga ttacttgatg ctgaaagtgg cccaccagcc tccatataca cagccccatt
     2821 gttctcctag acaaggccat gaactggcaa aacaagagat tcgagtgagg gttgaaaagg
     2881 atccagaact tggatttagc atatcaggtg gtgtcggggg tagaggaaac ccattcagac
     2941 ctgatgatga tggtatattt gtaacaaggg tacaacctga aggaccagca tcaaaattac
     3001 tgcagccagg tgataaaatt attcaggcta atggctacag ttttataaat attgaacatg
     3061 gacaagcagt gtccttgcta aaaactttcc agaatacagt tgaactcatc attgtacgag
     3121 aagtttcctc atgtacagac tacaaagacc atgacggtga ttataaagat catgacatcg
     3181 actacaagga tgacgatgac aagtaactcg agataatcaa cctctggatt acaaaatttg
     3241 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     3301 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     3361 taaatcctgg ttagttcttg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     3421 gacaggggct cggctgttgg gcactgacaa ttccgtggtg tttatttgtg aaatttgtga
     3481 tgctattgct ttatttgtaa ccatctagct ttatttgtga aatttgtgat gctattgctt
     3541 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     3601 tgtttcaggt tcagggggag atgtgggagg ttttttaaag atctggagac cggtgtggca
     3661 cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     3721 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     3781 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt
     3841 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     3901 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3961 cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4021 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     4081 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     4141 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     4201 ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga tttataaggg
     4261 attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4321 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     4381 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     4441 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     4501 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     4561 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     4621 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4681 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4741 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4801 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4861 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4921 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     4981 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5041 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5101 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5161 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5221 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5281 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5341 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5401 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5461 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5521 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5581 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5641 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5701 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5761 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5821 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5881 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     5941 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6001 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6061 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6121 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6181 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6241 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6301 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6361 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
