LOCUS       sequence-456645-        3480 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV backbone for Golden Gate assembly of donor templates using 
            BbsI/BpiI (overhangs AGCG and GCAA, respectively).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3480)
  AUTHORS   De La Cruz BM, Mitra S, He B, Celik M, Kaminski D, Smedler E, Sterky
            FH
  TITLE     Efficient Gene-Editing in Human Pluripotent Stem Cells Through 
            Simplified Assembly of Adeno-Associated Viral (AAV) Donor Templates.
  JOURNAL   Bio Protoc. 2024 Nov 5;14(21):e5097. doi: 10.21769/BioProtoc.5097. 
            eCollection 2024 Nov 5.
  PUBMED    39525974
REFERENCE   2  (bases 1 to 3480)
  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: "Bio 
            Protoc."; date: "2024-11-5"; volume: "14(21):e5097. doi"; pages: " 
            10.21769/BioProtoc.5097. eCollection 2024 Nov 5"
COMMENT     Sequence Label: pAAV-GG
FEATURES             Location/Qualifiers
     source          1..3480
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     repeat_region   743..883
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   743..872
                     /label=AAV2 ITR
     rep_origin      958..1413
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1045..1064)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1255..1276
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1430..1449)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1549..1571
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1609..1627)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1695..1799
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1800..1868,1869..2660)
                     /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(2018..2037)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2831..3419
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3320..3339
                     /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 gcgtgctctt cgcaagggtc ttctgagtta agggtgttgc
      181 tctcaatgat ttcattaatg gttcaatttt taatttcttt tcttctggtt taaaattcaa
      241 gtttaaagtg aaagtgtaat atgcacccat ttctttaaat aaatcttcta aatagtctac
      301 taatgtttta ttttgttttt tataaaatca agcagcctct gctattaata tagaagcttg
      361 tattccatct ttatctctag ctgagtcatc aattacatat ccataacttt cttcataagc
      421 aaaaacaaaa tttaatccgt tatcttcttc tttagcaatt tctctaccca ttcatttaaa
      481 tccagttaaa gtttttacaa tattaactcc atatttttca tgagcgattc tatcacccaa
      541 atcacttgtt acaaaacttg aatatagagc cggatttttt ggaatgctat ttaagcgttt
      601 tagatttgat aattttcaat caattaaaat tggtcctgtt tgatttccat ctaatcttac
      661 aaaatgacca tcatgtttta ttgccattcc aaatctgtca gcagaagaca tagcggtaac
      721 cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
      781 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
      841 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
      901 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
      961 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     1021 cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     1081 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     1141 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     1201 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     1261 tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt gatttataag
     1321 ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     1381 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     1441 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     1501 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     1561 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     1621 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     1681 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     1741 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     1801 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     1861 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     1921 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     1981 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     2041 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     2101 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     2161 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     2221 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     2281 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     2341 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     2401 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     2461 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     2521 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     2581 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     2641 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     2701 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     2761 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     2821 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     2881 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     2941 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     3001 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3061 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3121 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3181 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3241 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3301 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3361 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3421 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
