LOCUS       sequence-180625-        7572 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Construct used to synthesize progerin mRNA. Please note the plasmid 
            contains LMNA gene with nucleotide C1824T mutation..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7572)
  AUTHORS   Hewitt
  TITLE     pAAV-AcGFP-progerin
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7572)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-AcGFP-progerin
FEATURES             Location/Qualifiers
     source          1..7572
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        60..363
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        364..567
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     564..588
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          716..1094
                     /label=chimeric intron
                     /note="fusion between human cytomegalovirus intron A and 
                     human beta-globin intron 2"
     primer_bind     1183..1202
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1183..1201
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1242..1251
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1248..1964
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    4173..4649
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4372..4391)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4689..4829
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4689..4818
                     /label=AAV2 ITR
     rep_origin      4904..5359
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4991..5010)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5201..5222
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5376..5395)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5495..5517
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5555..5573)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5641..5745
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5746..5814,5815..6606)
                     /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(5964..5983)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6777..7365
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7266..7285
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   7427..7556
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
       61 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      121 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      181 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      241 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      301 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      361 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
      421 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      481 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
      541 cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc
      601 catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct ccgcggattc
      661 gaatcccggc cgggaacggt gcattggaac gcggattccc cgtgccaaga gtgacgtaag
      721 taccgcctat agagtctata ggcccacaaa aaatgctttc ttcttttaat atactttttt
      781 gtttatctta tttctaatac tttccctaat ctctttcttt cagggcaata atgatacaat
      841 gtatcatgcc tctttgcacc attctaaaga ataacagtga taatttctgg gttaaggcaa
      901 tagcaatatt tctgcatata aatatttctg catataaatt gtaactgatg taagaggttt
      961 catattgcta atagcagcta caatccagct accattctgc ttttatttta tggttgggat
     1021 aaggctggat tattctgagt ccaagctagg cccttttgct aatcatgttc atacctctta
     1081 tcttcctccc acagctcctg ggcaacgtgc tggtctgtgt gctggcccat cactttggca
     1141 aagaattggg attcgaacat cgattgaatt ccccggggat cctaatacga ctcactatag
     1201 gggaaataag agagaaaaga agagtaagaa gaaatataag agccaccatg gtgagcaagg
     1261 gcgccgagct gttcaccggc atcgtgccca tcctgatcga gctgaatggc gatgtgaatg
     1321 gccacaagtt cagcgtgagc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     1381 tgaagttcat ctgcaccacc ggcaagctgc ctgtgccctg gcccaccctg gtgaccaccc
     1441 tgagctacgg cgtgcagtgc ttctcacgct accccgatca catgaagcag cacgacttct
     1501 tcaagagcgc catgcctgag ggctacatcc aggagcgcac catcttcttc gaggatgacg
     1561 gcaactacaa gtcgcgcgcc gaggtgaagt tcgagggcga taccctggtg aatcgcatcg
     1621 agctgaccgg caccgatttc aaggaggatg gcaacatcct gggcaataag atggagtaca
     1681 actacaacgc ccacaatgtg tacatcatga ccgacaaggc caagaatggc atcaaggtga
     1741 acttcaagat ccgccacaac atcgaggatg gcagcgtgca gctggccgac cactaccagc
     1801 agaatacccc catcggcgat ggccctgtgc tgctgcccga taaccactac ctgtccaccc
     1861 agagcgccct gtccaaggac cccaacgaga agcgcgatca catgatctac ttcggcttcg
     1921 tgaccgccgc cgccatcacc cacggcatgg atgagctgta caagtccgga cttaaggcct
     1981 ctgtcgacag cagtctctgt ccttcgaccc gagccccgcg ccctttccgg gacccctgcc
     2041 ccgcgggcag cgctgccaac ctgccggcca tggagacccc gtcccagcgg cgcgccaccc
     2101 gcagcggggc gcaggccagc tccactccgc tgtcgcccac ccgcatcacc cggctgcagg
     2161 agaaggagga cctgcaggag ctcaatgatc gcttggcggt ctacatcgac cgtgtgcgct
     2221 cgctggaaac ggagaacgca gggctgcgcc ttcgcatcac cgagtctgaa gaggtggtca
     2281 gccgcgaggt gtccggcatc aaggccgcct acgaggccga gctcggggat gcccgcaaga
     2341 cccttgactc agtagccaag gagcgcgccc gcctgcagct ggagctgagc aaagtgcgtg
     2401 aggagtttaa ggagctgaaa gcgcgcaata ccaagaagga gggtgacctg atagctgctc
     2461 aggctcggct gaaggacctg gaggctctgc tgaactccaa ggaggccgca ctgagcactg
     2521 ctctcagtga gaagcgcacg ctggagggcg agctgcatga tctgcggggc caggtggcca
     2581 agcttgaggc agccctaggt gaggccaaga agcaacttca ggatgagatg ctgcggcggg
     2641 tggatgctga gaacaggctg cagaccatga aggaggaact ggacttccag aagaacatct
     2701 acagtgagga gctgcgtgag accaagcgcc gtcatgagac ccgactggtg gagattgaca
     2761 atgggaagca gcgtgagttt gagagccggc tggcggatgc gctgcaggaa ctgcgggccc
     2821 agcatgagga ccaggtggag cagtataaga aggagctgga gaagacttat tctgccaagc
     2881 tggacaatgc caggcagtct gctgagagga acagcaacct ggtgggggct gcccacgagg
     2941 agctgcagca gtcgcgcatc cgcatcgaca gcctctctgc ccagctcagc cagctccaga
     3001 agcagctggc agccaaggag gcgaagcttc gagacctgga ggactcactg gcccgtgagc
     3061 gggacaccag ccggcggctg ctggcggaaa aggagcggga gatggccgag atgcgggcaa
     3121 ggatgcagca gcagctggac gagtaccagg agcttctgga catcaagctg gccctggaca
     3181 tggagatcca cgcctaccgc aagctcttgg agggcgagga ggagaggcta cgcctgtccc
     3241 ccagccctac ctcgcagcgc agccgtggcc gtgcttcctc tcactcatcc cagacacagg
     3301 gtgggggcag cgtcaccaaa aagcgcaaac tggagtccac tgagagccgc agcagcttct
     3361 cacagcacgc acgcactagc gggcgcgtgg ccgtggagga ggtggatgag gagggcaagt
     3421 ttgtccggct gcgcaacaag tccaatgagg accagtccat gggcaattgg cagatcaagc
     3481 gccagaatgg agatgatccc ttgctgactt accggttccc accaaagttc accctgaagg
     3541 ctgggcaggt ggtgacgatc tgggctgcag gagctggggc cacccacagc ccccctaccg
     3601 acctggtgtg gaaggcacag aacacctggg gctgcgggaa cagcctgcgt acggctctca
     3661 tcaactccac tggggaagaa gtggccatgc gcaagctggt gcgctcagtg actgtggttg
     3721 aggacgacga ggatgaggat ggagatgacc tgctccatca ccaccacggc tcccactgca
     3781 gcagctcggg ggaccccgct gagtacaacc tgcgctcgcg caccgtgctg tgcgggacct
     3841 gcgggcagcc tgccgacaag gcatctgcca gcggctcagg agcccaggtg ggtggaccca
     3901 tctcctctgg ctcttctgcc tccagtgtca cggtcactcg cagctaccgc agtgtggggg
     3961 gcagtggggg tggcagcttc ggggacaatc tggtcacccg ctcctacctc ctgggcaact
     4021 ccagcccccg aacccagagc ccccagaact gcagcatcat gtaagctgcc ttctgcgggg
     4081 cttgccttct ggccatgccc ttcttctctc ccttgcacct gtacctcttg gtctttgaat
     4141 aaagcctgag taggaagtct cgagagatct acgggtggca tccctgtgac ccctccccag
     4201 tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat
     4261 taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg
     4321 tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg
     4381 aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc
     4441 aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc atgaccaggc
     4501 tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct
     4561 ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg
     4621 cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg tgcggaccga
     4681 gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     4741 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
     4801 gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca
     4861 tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc
     4921 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     4981 ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     5041 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     5101 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     5161 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     5221 ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     5281 tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     5341 atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     5401 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     5461 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     5521 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     5581 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     5641 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     5701 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     5761 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     5821 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     5881 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     5941 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     6001 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     6061 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     6121 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     6181 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     6241 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     6301 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     6361 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     6421 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     6481 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     6541 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     6601 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     6661 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6721 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6781 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6841 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6901 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     6961 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     7021 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     7081 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     7141 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     7201 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     7261 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     7321 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     7381 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtcctg caggcagctg
     7441 cgcgctcgct cgctcactga ggccgcccgg gcgtcgggcg acctttggtc gcccggcctc
     7501 agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg
     7561 ccgcacgcgt ct
//
