LOCUS       sequence-339579-        4270 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 4270)
  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 4270)
  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-EGFP
FEATURES             Location/Qualifiers
     source          1..4270
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        74..387
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     306..326
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     regulatory      477..486
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(483..485,486..488,489..1202)
                     /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(483..485,486..488,489..1199)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(528..549)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(789..808)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1136..1157
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1325..1446
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1362..1381)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1416..1435
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   1464..1593
                     /label=AAV2 ITR
     rep_origin      complement(1830..2418)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1910..1929)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2591..3382,3383..3451))
                     /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     3214..3233
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3452..3556)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3624..3642
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3680..3702)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3802..3821
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggaattcac
       61 gcgtatatcg atagatccga cgccgccatc tctaggcccg cgccggcccc ctcgcacaga
      121 cttgtgggag aagctcggct actcccctgc cccggttaat ttgcatataa tatttcctag
      181 taactataga ggcttaatgt gcgataaaag acagataatc tgttcttttt aatactagct
      241 acattttaca tgataggctt ggatttctat aagagataca aatactaaat tattatttta
      301 aaaaacagca caaaaggaaa ctcaccctaa ctgtaaagta attgtgtgtt ttgagactat
      361 aaatatccct tggagaaaag ccttgtttgg gcccggccac aagttcagcg tgttcaagag
      421 acacgctgaa cttgtggccg ttttttaatt cctgcagccc gggggatcca ccggtcgcca
      481 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
      541 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
      601 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
      661 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
      721 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
      781 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
      841 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
      901 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
      961 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     1021 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     1081 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     1141 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     1201 aaagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc
     1261 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     1321 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     1381 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     1441 atcttaaggc gggaattgat ctgaggaacc cctagtgatg gagttggcca ctccctctct
     1501 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc
     1561 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcatt aatgaatcgg ccaacgcgcg
     1621 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     1681 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     1741 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     1801 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     1861 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     1921 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     1981 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     2041 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2101 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2161 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2221 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     2281 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     2341 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     2401 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     2461 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     2521 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatat gagtaaactt
     2581 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     2641 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     2701 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     2761 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     2821 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     2881 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     2941 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     3001 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     3061 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     3121 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     3181 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     3241 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     3301 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     3361 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     3421 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     3481 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     3541 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     3601 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
     3661 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     3721 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     3781 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     3841 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcga ttccaacatc
     3901 caataaatca tacaggcaag gcaaagaatt agcaaaatta agcaataaag cctcagagca
     3961 taaagctaaa tcggttgtac caaaaacatt atgaccctgt aatacttttg cgggagaagc
     4021 ctttatttca acgcaaggat aaaaattttt agaaccctca tatattttaa atgcaatgcc
     4081 tgagtaatgt gtaggtaaag attcaaacgg gtgagaaagg ccggagacag tcaaatcacc
     4141 atcaatatga tattcaaccg ttctagctga taaattcatg ccggagaggg tagctatttt
     4201 tgagaggtct ctacaaaggc tatcaggtca ttgcctgaga gtctggagca aacaagagaa
     4261 tcgcctgcag
//
