LOCUS       sequence-9621-ex        5416 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5416)
  AUTHORS   Mejat A, Decostre V, Li J, Renou L, Kesari A, Hantai D, Stewart CL, 
            Xiao X, Hoffman E, Bonne G, Misteli T
  TITLE     Lamin A/C-mediated neuromuscular junction defects in Emery-Dreifuss 
            muscular dystrophy.
  JOURNAL   J Cell Biol. 2009 Jan 12. 184(1):31-44.
  PUBMED    19124654
REFERENCE   2  (bases 1 to 5416)
  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: "J Cell 
            Biol. 2009 Jan 12. 184(1):31-44."
COMMENT     Sequence Label: pEMBL-Si-LMNA
FEATURES             Location/Qualifiers
     source          1..5416
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      423..936
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(510..529)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     720..741
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1142..1158
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1155..1468
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     1387..1407
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     complement(1546..1562)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      1581..1590
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1587..1589,1590..1592,1593..2306)
                     /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             join(1587..1589,1590..1592,1593..2303)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1632..1653)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1893..1912)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2240..2261
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2355..2372)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2361..2568
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3194..3213)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3313..3335
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3373..3391)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3459..3563
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3564..3632,3633..4424)
                     /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(3782..3801)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4595..5183
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5084..5103
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cagcagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
       61 cctgaatggc gaatggaatt ccagacgatt gagcgtcaaa atgtaggtat ttccatgagc
      121 gtttttcctg ttgcaatggc tggcggtaat attgttctgg atattaccag caaggccgat
      181 agtttgagtt cttctactca ggcaagtgat gttattacta atcaaagaag tattgcgaca
      241 acggttaatt tgcgtgatgg acagactctt ttactcggtg gcctcactga ttataaaaac
      301 acttctcagg attctggcgt accgttcctg tctaaaatcc ctttaatcgg cctcctgttt
      361 agctcccgct ctgattctaa cgaggaaagc acgttatacg tgctcgtcaa agcaaccata
      421 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
      481 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
      541 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
      601 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
      661 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
      721 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
      781 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
      841 taacgcgaat tttaacaaaa tattaacgtt tacaatttaa atatttgctt atacaatctt
      901 cctgtttttg gggcttttct gattatcaac cggggtacat atgattgaca tgctagtttt
      961 acgattaccg ttcatcgcct gcactgcgcg ctcgctcgct cactgaggcc gcccgggcaa
     1021 agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag
     1081 agggagtgga attcacgcgt ggtacgatct gaattcggta caattcacgc gtggtacccg
     1141 ctctagaact agtggatccg acgccgccat ctctaggccc gcgccggccc cctcgcacag
     1201 acttgtggga gaagctcggc tactcccctg ccccggttaa tttgcatata atatttccta
     1261 gtaactatag aggcttaatg tgcgataaaa gacagataat ctgttctttt taatactagc
     1321 tacattttac atgataggct tggatttcta taagagatac aaatactaaa ttattatttt
     1381 aaaaaacagc acaaaaggaa actcacccta actgtaaagt aattgtgtgt tttgagacta
     1441 taaatatccc ttggagaaaa gccttgtttg ggccggagct tgacttccag aagttcaaga
     1501 gacttctgga agtcaagctc cttttttaat tcctgcagcc cgggggatcc actagttcta
     1561 gagcggccga tccaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg
     1621 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     1681 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     1741 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     1801 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     1861 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     1921 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     1981 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     2041 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     2101 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     2161 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     2221 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     2281 tcggcatgga cgagctgtac aagtaaagcg gccatcaagc ttatcgatac cgtcgactag
     2341 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     2401 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     2461 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     2521 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggaga gatcgatcta
     2581 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     2641 cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg
     2701 agcgcgcaga gagggagtgg ccaacccccc cccccccccc cctgcatgca ggcgattctc
     2761 ttgtttgctc cagactctca ggcaatgacc tgatagcctt tgtagagacc tctcaaaaat
     2821 agctaccctc tccggcatga atttatcagc tagaacggtt gaatatcata ttgatggtga
     2881 tttgactgtc tccggccttt ctcacccgtt tgaatcttta cctacacatt actcaggcat
     2941 tgcatttaaa atatatgagg gttctaaaaa tttttatcct tgcgttgaaa taaaggcttc
     3001 tcccgcaaaa gtattacagg gtcataatgt ttttggtaca accgatttag ctttatgctc
     3061 tgaggcttta ttgcttaatt ttgctaattc tttgccttgc ctgtatgatt tattggatgt
     3121 tggaattcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat
     3181 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     3241 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     3301 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     3361 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     3421 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3481 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3541 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     3601 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     3661 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     3721 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     3781 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     3841 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     3901 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     3961 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     4021 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     4081 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     4141 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     4201 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     4261 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     4321 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     4381 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     4441 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     4501 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     4561 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4621 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4681 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     4741 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     4801 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     4861 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     4921 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     4981 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5041 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5101 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     5161 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     5221 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     5281 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     5341 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     5401 ggccgattca ttaatg
//
