LOCUS       sequence-73245-e        6226 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV plasmid with smCBA promoter driving expression of iCre. Contains
            WPRE..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6226)
  AUTHORS   Mirjalili Mohanna SZ, Korecki AJ, Simpson EM
  TITLE     rAAV-PHP.B escapes the mouse eye and causes lethality whereas rAAV9 
            can transduce aniridic corneal limbal stem cells without lethality.
  JOURNAL   Gene Ther. 2023 Apr 19. doi: 10.1038/s41434-023-00400-6.
  PUBMED    37072572
REFERENCE   2  (bases 1 to 6226)
  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: "Gene Ther. 
            2023 Apr 19. doi: 10.1038/s41434-023-00400-6."
COMMENT     Sequence Label: pEMS2118
FEATURES             Location/Qualifiers
     source          1..6226
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     enhancer        276..579
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        581..858
                     /label=chicken beta-actin promoter
     primer_bind     1115..1134
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          1190..1322
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1305..1324
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     complement(2446..2466)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2864..2875)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2996..3117
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3007..3026)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3061..3080
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3287..3416
                     /label=AAV2 ITR
     primer_bind     complement(3435..3452)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3435..3451)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3444..3466)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3593..4048
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3680..3699)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3890..3911
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4074..4178
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4179..4247,4248..5039)
                     /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(4397..4416)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5210..5798
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5699..5718
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6086..6107
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6122..6127,6128..6145,6146..6152)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6160..6176
                     /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)."
     primer_bind     6165..6187
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6184..6200
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6184..6200
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagatcg gaattcccta ggccggccgg taccctagtt attaatagta atcaattacg
      241 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
      301 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
      361 atagtaacgc caatagggac tttccattga cgtcaatggg tggactattt acggtaaact
      421 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
      481 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
      541 tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc
      601 tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt
      661 taattatttt gtgcagcgat gggggcgggg gggggggggg ggcgcgcgcc aggcggggcg
      721 gggcggggcg aggggcgggg cggggcgagg cggagaggtg cggcggcagc caatcagagc
      781 ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag
      841 cgaagcgcgc ggcgggcggg agtcgctgcg acgctgcctt cgccccgtgc cccgctccgc
      901 cgccgcctcg cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg
      961 gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc
     1021 ttttctgtgg ctgcgtgaaa gccttgaggg gctccgggag ctagagcctc tgctaaccat
     1081 gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat tgtgctgtct
     1141 catcattttg gcaaaggcgc gccgaagttg gtcgtgaggc actgggcagg taagtatcaa
     1201 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
     1261 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1321 aggtgtccac tcccaggcgg ccgccaccat gtccaacctg ctgactgtgc accaaaacct
     1381 gcctgccctc cctgtggatg ccacctctga tgaagtcagg aagaacctga tggacatgtt
     1441 cagggacagg caggccttct ctgaacacac ctggaagatg ctcctgtctg tgtgcagatc
     1501 ctgggctgcc tggtgcaagc tgaacaacag gaaatggttc cctgctgaac ctgaggatgt
     1561 gagggactac ctcctgtacc tgcaagccag aggcctggct gtgaagacca tccaacagca
     1621 cctgggccag ctcaacatgc tgcacaggag atctggcctg cctcgccctt ctgactccaa
     1681 tgctgtgtcc ctggtgatga ggagaatcag aaaggagaat gtggatgctg gggagagagc
     1741 caagcaggcc ctggcctttg aacgcactga ctttgaccaa gtcagatccc tgatggagaa
     1801 ctctgacaga tgccaggaca tcaggaacct ggccttcctg ggcattgcct acaacaccct
     1861 gctgcgcatt gccgaaattg ccagaatcag agtgaaggac atctcccgca ccgatggtgg
     1921 gagaatgctg atccacattg gcaggaccaa gaccctggtg tccacagctg gtgtggagaa
     1981 ggccctgtcc ctgggggtta ccaagctggt ggagagatgg atctctgtgt ctggtgtggc
     2041 tgatgacccc aacaactacc tgttctgccg ggtcagaaag aatggtgtgg ctgccccttc
     2101 tgccacctcc caactgtcca cccgggccct ggaagggatc tttgaggcca cccaccgcct
     2161 gatctatggt gccaaggatg actctgggca gagatacctg gcctggtctg gccactctgc
     2221 cagagtgggt gctgccaggg acatggccag ggctggtgtg tccatccctg aaatcatgca
     2281 ggctggtggc tggaccaatg tgaacatagt gatgaactac atcagaaacc tggactctga
     2341 gactggggcc atggtgaggc tgctcgagga tggggactga gcggccgcga tcgctcaacc
     2401 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     2461 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     2521 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     2581 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     2641 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     2701 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     2761 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     2821 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     2881 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     2941 ccctcagacg agtcggatct ccctttgggc cgcctccccg cgatcgcaga catgataaga
     3001 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     3061 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     3121 aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa
     3181 agcaagtaaa acctctacaa atgtggtagg tcgaaattcc cgataaggat cttcctagag
     3241 catggctacg tagataagta gcatggcggg ttaatcatta actacaagga acccctagtg
     3301 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     3361 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagcctt
     3421 aattaaccta attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     3481 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     3541 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc
     3601 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3661 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3721 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3781 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     3841 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3901 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     3961 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4021 tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa
     4081 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4141 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     4201 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     4261 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     4321 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     4381 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     4441 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     4501 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     4561 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     4621 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     4681 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     4741 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     4801 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     4861 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     4921 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     4981 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     5041 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     5101 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     5161 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     5221 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     5281 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     5341 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     5401 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     5461 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     5521 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5581 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     5641 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     5701 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     5761 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     5821 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     5881 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     5941 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     6001 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     6061 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     6121 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     6181 acacaggaaa cagctatgac catgattacg ccagatttaa ttaagg
//
