LOCUS       sequence-64892-e        6968 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 6968)
  AUTHORS   Metzger T, Gache V, Xu M, Cadot B, Folker ES, Richardson BE, Gomes 
            ER, Baylies MK
  TITLE     MAP and kinesin-dependent nuclear positioning is required for 
            skeletal muscle function.
  JOURNAL   Nature. 2012 Mar 18;484(7392):120-4. doi: 10.1038/nature10914.
  PUBMED    22425998
REFERENCE   2  (bases 1 to 6968)
  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"; doi: "10"; journalName: 
            "Nature"; date: "2012-03-18- 18"; volume: "484"; issue: "7392"; 
            pages: "120-4"
COMMENT     Sequence Label: pEGFP-Map7-fulllength
FEATURES             Location/Qualifiers
     source          1..6968
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3756..3877
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3793..3812)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3847..3866
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3884..4339)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4021..4042)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4233..4252
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4366..4470
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4472..4829
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4472..4663
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4495..4515)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4680..4815
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4864..5658
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4864..5658
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4918..4937)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5528..5547
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5846..5865)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5890..5937
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6266..6854
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6755..6774
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagct caagcttcga attcactagt gattatggcg
     1381 gagcagggag ctggcggcga cggccacagg ggcggcgacg gcgctacgca cagcgaccca
     1441 gcgtctgatg gctataaagt acaagagaag agaactgccc ccagccgccc cacttctaca
     1501 gtttcaggac aaactagcaa ccactcagga aacaaaccag atcccccacc tgtgctacgg
     1561 gttgatgacc ggcagcggct agccagagaa cgccgagagg aacgagaaaa acagctagct
     1621 gcacgagaaa ccgtctggct agagagagaa gaacgagccc ggcagcacta tgagagacat
     1681 ctggaggcga ggaagaagaa gctggaggac caacggctga aggaggagcg gaggagggct
     1741 gcggtggagg agaagcggag gcagaggctg gaggaggaca aggagcgcca tgaagctgtt
     1801 gtccgacgga cgatggaaag gagccagaag ccaaggcaga agtctaaccg ctggtcctgg
     1861 ggaagccctc tccatgggag ctcgagcatc cacagtggag atccagacag gcgctcagtt
     1921 tccaccatga atctttcgaa acatgttgat cctgtcatta gcaagcggct ctcctcctcg
     1981 tctgcaactt tgctaaactc tccagataga gctcgccgcc tgcagctcag cccctgggag
     2041 agcagcgttg ttagcagact tctgacgccc acacattcgt tcctggccag aagcaaaagc
     2101 acggccgcct tgtctggaga cacagttatc cccatttgtc ctcgttcagc atcttgcagc
     2161 cccatcatca tgcccttcaa agctgcacac tctagaaacc cagtggaccg accaaaactc
     2221 tttgtaacac cgcctgaggg ctctgcacga aggaggacca ttcatggact agcgagccat
     2281 aaaagagagc gagaaagaga acacgttccc ttccacgttt ccccgggcgc ccgcaggact
     2341 ctgtctccat ctaatctcaa agcgaggtca ccggctccag cccgcctttg gctcccatcc
     2401 aagtccatgc ctcatctgcc tggtactccc cggcctgcat cctccttgcc tcccggctca
     2461 gtcagagctg cttccgctca ggccccctcc tcctctcctg gcaacatccg gcctgtcaag
     2521 agagaagtga aagtggagcc tgagaagaaa gaccctttac ccgcagtaaa gagcagggtg
     2581 ccattagtga aggtagagga ggtcacagtc gaagagggga cacccgtgaa gccacctgag
     2641 cctgctgctc cagcctcggc ccccattgca accccagccc ctgctccagc tacggacccg
     2701 gccccagtcc ctgcaccatc atccactgtg actgtcggtg tagttcctaa gacttctgca
     2761 ggcaccaccg acccagagga ggctacgagg ttgctggctg agaagaggcg tctagccaga
     2821 gagcagaggg agaaggagga acgggagagg aaggaaaagg aagagctgga gagacaaaag
     2881 atagaggaat tggcccgtag ggtggctgaa gagcgaagtc gcagggaaga agaagcccgc
     2941 aggctggaag aggagcaggc tcgagagaaa gaggagctgg cgctgcgtct ggctgaggag
     3001 gagcgggagc ggtgggagag ggaggaggtg gagcgcgtgc agaagcagaa agaagaagaa
     3061 gcccgagccc gggaggaggc agagagggct cggcaggaac gagagaagca tttccagaaa
     3121 gaagagcagg aacggctgga gaggaagaag cgacttgaag agattatgag aagaaccagg
     3181 aggacagaga ccgctgataa gaaaaccact gagcaaagaa atggtgacat agccaaagga
     3241 gttctcactg gagagccaga agtacctgca ctgccgtgta tggcctcttc aggaaacgga
     3301 gagtctgcag agagcccaca tggagtcgct ttacagcaat cagaagtgac cacagagagt
     3361 tctccagatt tggaaaaaca gccaaatgaa aacggaatgt ccatacaaaa tgaaaatttt
     3421 gaagaagtta taaacttacc tgttggatca aaagcgtcca gattagatgt caccaatgag
     3481 aacccagaaa ttcctttgaa accaattttg gcctttaatg atgaagggac acttgggccc
     3541 ctacctcagg tggatggtgt gcagacacaa cagaccgcag aagttaatca ctagtgaatt
     3601 ctgcagtcga cggtaccgcg ggcccgggat ccaccggatc tagataactg atcataatca
     3661 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     3721 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     3781 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     3841 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat
     3901 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     3961 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     4021 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     4081 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     4141 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     4201 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     4261 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     4321 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4381 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4441 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     4501 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4561 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4621 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4681 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4741 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     4801 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     4861 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4921 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4981 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     5041 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     5101 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     5161 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     5221 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     5281 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     5341 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     5401 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     5461 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     5521 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     5581 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     5641 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     5701 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     5761 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5821 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     5881 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     5941 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     6001 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     6061 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     6121 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     6181 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6241 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     6301 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6361 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     6421 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     6481 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6541 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6601 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6661 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6721 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6781 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6841 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6901 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     6961 ccatgcat
//
