LOCUS       sequence-421849-        4323 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Self-complementary AAV vector packaging GFP under the human Iba1 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4323)
  AUTHORS   Serrano C, Cananzi S, Shen T, Wang LL, Zhang CL
  TITLE     Simple and highly specific targeting of resident microglia with 
            adeno-associated virus.
  JOURNAL   iScience. 2024 Aug 12;27(9):110706. doi: 10.1016/j.isci.2024.110706.
            eCollection 2024 Sep 20.
  PUBMED    39297168
REFERENCE   2  (bases 1 to 4323)
  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: "iScience.";
            date: "2024-08-12"; volume: "27(9):110706. doi"; pages: " 
            10.1016/j.isci.2024.110706. eCollection 2024 Sep 20"
COMMENT     Sequence Label: scAAV-hIBA1a-GFP
FEATURES             Location/Qualifiers
     source          1..4323
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..27)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11..27)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(24..46)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    35..51
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(59..65,66..83,84..89))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    104..125
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(413..1001)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(493..512)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1172..1963,1964..2032))
                     /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     1795..1814
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2033..2137)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2205..2223
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2261..2283)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2383..2402
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2596..2618
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2610..2627
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2611..2627
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2877..2896)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(2940..3653,3654..3656,3657..3659))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(2943..3653,3654..3656,3657..3659))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2985..3006)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3334..3353
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3593..3614
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3656..3665
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
       61 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
      121 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
      181 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
      241 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
      301 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
      361 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
      421 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
      481 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
      541 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
      601 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
      661 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
      721 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
      781 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
      841 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
      901 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
      961 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     1021 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     1081 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     1141 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     1201 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     1261 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     1321 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     1381 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     1441 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     1501 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     1561 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     1621 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     1681 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     1741 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     1801 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     1861 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     1921 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     1981 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     2041 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     2101 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     2161 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     2221 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     2281 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     2341 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     2401 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     2461 gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     2521 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     2581 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttgtcgacgc
     2641 atgcgggccc ggatctgtcc ctctctgcgc gctcgctcgc tcactgaggc cgcccgggca
     2701 aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga
     2761 gagggagtgg actagtttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     2821 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     2881 tagcatcaca aatttcacaa ataaagcatc gagatcgcag gtgaggccta gcggccgctt
     2941 tacttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg
     3001 accatgtgat cgcgcttctc gttggggtct ttgctcaggg cggactgggt gctcaggtag
     3061 tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg
     3121 gcgagctgca cgctgccgtc ctcgatgttg tggcggatct tgaagttcac cttgatgccg
     3181 ttcttctgct tgtcggccat gatatagacg ttgtggctgt tgtagttgta ctccagcttg
     3241 tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc
     3301 agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc gtccttgaag
     3361 aagatggtgc gctcctggac gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc
     3421 ttcatgtggt cggggtagcg gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg
     3481 gtgggccagg gcacgggcag cttgccggtg gtgcagatga acttcagggt cagcttgccg
     3541 taggtggcat cgccctcgcc ctcgccggac acgctgaact tgtggccgtt tacgtcgccg
     3601 tccagctcga ccaggatggg caccaccccg gtgaacagct cctcgccctt gctcaccatg
     3661 gtggcgaccg gtagctcagc agatgctggt agaggtgggg agacagacca agctggaggc
     3721 ctctgtctgc aggctctcct tctctcccac cagtcttctc agaagccttc ctcccaaact
     3781 tcccctttcc cctcattcct tcctcctctg cctcccagac ctcacttccc catcaggttc
     3841 tgctgacact agcaccccag gaaggagggg gagcctggtg tgaaatgcag attagaaaat
     3901 aacattctcc caaattcctc ttaaatgaga gcccagtgtg cgcatgaaac aaatattttg
     3961 ataagccact ggaggtgtgc aggaagggca ggaagacacc agggatattt attctcagga
     4021 actagtattt ttgaacaggt ccaaggttca ggtaatgata agaccttgca ttttgtcagg
     4081 tttcatgagt ctgggcaggc agctgtggat aggaagacaa ggaagggctg gctgaatttc
     4141 taggtcacga ctccacccct ccaggaaccc ctagtgatgg agttggccac tccctctctg
     4201 cgcgctcgct cgctcactga ggccgggcga cctttggtcg cccggcctca gtgagcgagc
     4261 gagcgcgcag agagggagtg gcccagatcc ccccgggaag ctttctagat ctcgagagct
     4321 tgg
//
