LOCUS       sequence-407719-        5559 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Beta cell-specific gene expression from AAV in human pancreatic 
            islets.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5559)
  AUTHORS   Chai S, Kim Y, Galivo F, Dorrell C, Wakefield L, Posey J, Ackermann 
            AM, Kaestner KH, Hebrok M, Grompe M
  TITLE     Development of a Beta Cell-Specific Expression Control Element for 
            Recombinant Adeno-Associated Virus.
  JOURNAL   Hum Gene Ther. 2022 Aug;33(15-16):789-800. doi: 
            10.1089/hum.2021.219. Epub 2022 May 10.
  PUBMED    35297680
REFERENCE   2  (bases 1 to 5559)
  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: "Hum Gene 
            Ther."; date: "2022-08"; volume: "33(15-16):789-800. doi"; pages: " 
            10.1089/hum.2021.219. Epub 2022 May 10"
COMMENT     Sequence Label: pAAV INS2x mRFP-3MREs (AAV-beta)
FEATURES             Location/Qualifiers
     source          1..5559
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(125..229)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(255..710)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(392..413)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     604..623
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   782..911
                     /label=AAV2 ITR
     regulatory      1830..1839
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1836..2513
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             1836..2513
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(1966..1984)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2234..2253
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2428..2451
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    2784..3372
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2837..2857)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3255..3266)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3415..3536
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3426..3445)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3480..3499
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3698..3827
                     /label=AAV2 ITR
     rep_origin      complement(4064..4652)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4144..4163)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4823..5559,1..55,56..124))
                     /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     5446..5465
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
       61 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
      121 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
      181 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
      241 gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
      301 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaattaaa
      361 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
      421 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
      481 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
      541 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
      601 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
      661 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc
      721 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
      781 gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt
      841 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
      901 taggggttcc ttgtagttaa tgattaaccc gccatgctac ttatctacgt agccatgctc
      961 taggaagagt accattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     1021 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     1081 aagtgtatca tatgagatgg gctctgagac ctcgagcagc agcgcaaaga gccccgccct
     1141 gcagcctcca gctctcctgg tctaatgtgg aaagtggccc aggtgagggc tttgctctcc
     1201 tggagacatt tgcccccagc tgtgagcagg gacaggtctg gccaccgggc ccctggttaa
     1261 gactctaatg acccgctggt cctgaggaag aggtgctgac gaccaaggag atcttcccac
     1321 agacccagca ccagggaaat ggtccggaaa ttgcagcctc agcccccagc catctgccga
     1381 cccccccacc ccaggcccta atgggccagg cggcaggggt tgagaggtag gggagatggg
     1441 ctctgagaca agcttcagca gcgcaaagag ccccgccctg cagcctccag ctctcctggt
     1501 ctaatgtgga aagtggccca ggtgagggct ttgctctcct ggagacattt gcccccagct
     1561 gtgagcaggg acaggtctgg ccaccgggcc cctggttaag actctaatga cccgctggtc
     1621 ctgaggaaga ggtgctgacg accaaggaga tcttcccaca gacccagcac cagggaaatg
     1681 gtccggaaat tgcagcctca gcccccagcc atctgccgac ccccccaccc caggccctaa
     1741 tgggccaggc ggcaggggtt gagaggtagg ggagatgggc tctgagacta taaagccagc
     1801 gggggcccag cagccctcgc tagccggtcg ccaccatggc ctcctccgag gacgtcatca
     1861 aggagttcat gcgcttcaag gtgcgcatgg agggctccgt gaacggccac gagttcgaga
     1921 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga
     1981 ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc cagtacggct
     2041 ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg
     2101 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc
     2161 aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact
     2221 tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc tccaccgagc
     2281 ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gatgaggctg aagctgaagg
     2341 acggcggcca ctacgacgcc gaggtcaaga ccacctacat ggccaagaag cccgtgcagc
     2401 tgcccggcgc ctacaagacc gacatcaagc tggacatcac ctcccacaac gaggactaca
     2461 ccatcgtgga acagtacgag cgcgccgagg gccgccactc caccggcgcc taaatgcata
     2521 agctgtacaa ctcaccgaca gcgttgaatg ttactcaccg acagcgttga atgttactca
     2581 ccgacagcgt tgaatgttaa gcttaagctg tacaactcac acctaggttc caaggattac
     2641 tcacacctag gttccaagga ttactcacac ctaggttcca aggattaagc ttaagctgta
     2701 cacccagcta gattgtaagc tccttcccag ctagattgta agctccttcc cagctagatt
     2761 gtaagctcct taagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga
     2821 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     2881 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     2941 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     3001 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     3061 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     3121 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     3181 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     3241 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     3301 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     3361 ccgcctcccc gcatcgatac cgtcgacccg ggcggccgct tcgagcagac atgataagat
     3421 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     3481 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     3541 acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa
     3601 gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga tcttcctaga gcatggctac
     3661 gtagataagt agcatggcgg gttaatcatt aactacaagg aacccctagt gatggagttg
     3721 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     3781 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cattaatgaa
     3841 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     3901 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3961 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4021 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4081 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4141 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4201 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4261 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4321 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4381 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4441 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4501 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4561 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4621 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4681 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4741 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4801 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4861 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4921 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4981 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5041 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5101 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5161 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5221 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5281 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5341 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5401 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5461 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5521 ggatcttacc gctgttgaga tccagttcga tgtaaccca
//
