LOCUS       sequence-407724-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Alpha 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 5131)
  TITLE     AAV-alpha
REFERENCE   2  (bases 1 to 5131)
  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"
COMMENT     Sequence Label: pAAV MP-135 mRFP-3MREs (AAV-alpha)
FEATURES             Location/Qualifiers
     source          1..5131
                     /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
     intron          1278..1410
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1393..1412
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1447..1456
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1453..2130
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             1453..2130
                     /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(1583..1601)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1851..1870
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2045..2068
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    2356..2944
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2409..2429)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2827..2838)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2987..3108
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2998..3017)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3052..3071
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3270..3399
                     /label=AAV2 ITR
     rep_origin      complement(3636..4224)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3716..3735)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4395..5131,1..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     5018..5037
                     /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 ataaatcaaa
      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 tatgacgcgt gaattccatt tgtaacaaaa actcattatt tacagatgag
     1141 aaatttatat tgtcagcgta atatctgtga ggctaaacag agctggagag tatataaaag
     1201 cagtgcgcct tggtgcagaa gtacagagct taggacacag agctagctgc agaagttggt
     1261 cgtgaggcac tgggcaggta agtatcaagg ttacaagaca ggtttaagga gaccaataga
     1321 aactgggctt gtcgagacag agaagactct tgcgtttctg ataggcacct attggtctta
     1381 ctgacatcca ctttgccttt ctctccacag gtgtccactc ccagttcaat tcactgtgca
     1441 ccggtcgcca ccatggcctc ctccgaggac gtcatcaagg agttcatgcg cttcaaggtg
     1501 cgcatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
     1561 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggcggccc cctgcccttc
     1621 gcctgggaca tcctgtcccc tcagttccag tacggctcca aggcctacgt gaagcacccc
     1681 gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
     1741 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc
     1801 gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg
     1861 cagaagaaga ccatgggctg ggaggcctcc accgagcgga tgtaccccga ggacggcgcc
     1921 ctgaagggcg agatcaagat gaggctgaag ctgaaggacg gcggccacta cgacgccgag
     1981 gtcaagacca cctacatggc caagaagccc gtgcagctgc ccggcgccta caagaccgac
     2041 atcaagctgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgagcgc
     2101 gccgagggcc gccactccac cggcgcctaa atgcatagcc aagctcagac ggatccgaag
     2161 ccaagctcag acggatccga agccaagctc agacggatcc gaaagcttcg tacgctatac
     2221 ggtctactac gtacgctata cggtctacta cgtacgctat acggtctact actcgagagt
     2281 gaattctacc agtgccataa gtgaattcta ccagtgccat aagtgaattc taccagtgcc
     2341 atatctagaa tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     2401 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     2461 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     2521 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     2581 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     2641 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     2701 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     2761 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     2821 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     2881 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     2941 ccgcatcgat accgtcgacc cgggcggccg cttcgagcag acatgataag atacattgat
     3001 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     3061 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     3121 tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggtttttta aagcaagtaa
     3181 aacctctaca aatgtggtaa aatcgataag gatcttccta gagcatggct acgtagataa
     3241 gtagcatggc gggttaatca ttaactacaa ggaaccccta gtgatggagt tggccactcc
     3301 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     3361 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcattaatg aatcggccaa
     3421 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     3481 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3541 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3601 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3661 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3721 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3781 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3841 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3901 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3961 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4021 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4081 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4141 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4201 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4261 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     4321 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4381 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4441 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4501 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4561 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4621 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4681 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     4741 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4801 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     4861 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     4921 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4981 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5041 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5101 ccgctgttga gatccagttc gatgtaaccc a
//
