LOCUS       sequence-64893-e        7351 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 7351)
  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 7351)
  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-Map7emtb-Kif5bmotor
FEATURES             Location/Qualifiers
     source          1..7351
                     /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    4139..4260
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4176..4195)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4230..4249
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4267..4722)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4404..4425)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4616..4635
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4749..4853
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4855..5212
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4855..5046
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4878..4898)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5063..5198
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5247..6041
                     /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             5247..6041
                     /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(5301..5320)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5911..5930
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6229..6248)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6273..6320
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6649..7237
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7138..7157
                     /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 ccggaatggc ggacctggcc gagtgcaaca tcaaagtgat gtgtcgcttc
     1381 agacctctca acgagtctga agtgaaccgc ggcgacaagt acatcgccaa gtttcaggga
     1441 gaagacacgg tcgtgatcgc gtccaagcct tatgcatttg atcgggtgtt ccagtcaagc
     1501 acatctcaag agcaagtgta taatgactgt gcaaagaaga ttgttaaaga tgtacttgaa
     1561 ggatataatg gaacaatatt tgcatacggc caaacatcct ctgggaagac acacacaatg
     1621 gagggtaaac ttcatgatcc agaaggcatg ggaattattc caagaatagt gcaagatatt
     1681 tttaattata tttactccat ggatgaaaat ttggaatttc atattaaggt ttcatatttt
     1741 gaaatatatt tggataagat aagggacctg ttagatgttt caaagaccaa cctttcagtt
     1801 catgaagaca aaaaccgagt tccctatgta aaggggtgca cagagcgttt tgtatgtagt
     1861 ccagatgaag ttatggatac catagatgaa ggaaaatcca acagacatgt agcagttaca
     1921 aatatgaatg aacatagctc taggagtcac agtatatttc ttattaatgt caaacaagag
     1981 aacacacaaa cggaacaaaa gctgagtgga aaactttatc tagttgattt agctggtagt
     2041 gaaaaggtta gtaaaactgg agctgaaggt gctgtgctgg atgaagctaa aaacatcaac
     2101 aagtcacttt ctgctcttgg aaatgttatt tctgctttgg ctgagggtag tacttatgtt
     2161 ccatatcgag atagtaaaat gacaagaatc cttcaagatt cattaggtgg caactgtaga
     2221 accactattg taatttgctg ctctccatca tcatacaatg agtctgaaac aaaatctaca
     2281 ctcttatttg gccaaagggc caaaacaatt aagaacacag tttgtgtcaa tgtggagtta
     2341 actgcagaac agtggaaaaa gaagtatgaa aaagaaaaag aaaaaaataa gatcctgcgg
     2401 aacactattc agtggcttga aaatgagctc aacagatggc gtaatgggga gacggtgcct
     2461 attgatgaac agtttgacaa agagaaagcc aacttggaag ctttcacagt ggataaagat
     2521 attactctta ccaatgataa accagcaacc gcaattggag ttataggaaa ttttactgat
     2581 gctgaaagaa gaaagtgtga agaagaaatt gctaaattat acaaacagct tgatgacaag
     2641 gatgaagaaa ttaaccagca aagtcaactg gtagagaagc tgaagacgca aatgttggat
     2701 caggaggagc ttttggcatc taccagaagg gatcaagaca atatgcaagc tgagctgaat
     2761 cgccttcaag cagaaaatga tgcctctaaa gaagaagtga aagaagtttt acaggcccta
     2821 gaagaacttg ctgtcaatta tgatcagaag tctcaggaag ttgaagacaa aactaaggaa
     2881 tatgaattgc ttagtgatga attgaatcag aaatcggcaa ctttagcgag tatagatgct
     2941 gagcttcaga aacttaagga aatgaccaac caccagaaaa aacgagcagc tgagatgatg
     3001 gcatctttac taaaaggtac cagatctcga gctcaagctt cgaattcact agtgattatg
     3061 gcggagcagg gagctggcgg cgacggccac aggggcggcg acggcgctac gcacagcgac
     3121 ccagcgtctg atggctataa agtacaagag aagagaactg cccccagccg ccccacttct
     3181 acagtttcag gacaaactag caaccactca ggaaacaaac cagatccccc acctgtgcta
     3241 cgggttgatg accggcagcg gctagccaga gaacgccgag aggaacgaga aaaacagcta
     3301 gctgcacgag aaaccgtctg gctagagaga gaagaacgag cccggcagca ctatgagaga
     3361 catctggagg cgaggaagaa gaagctggag gaccaacggc tgaaggagga gcggaggagg
     3421 gctgcggtgg aggagaagcg gaggcagagg ctggaggagg acaaggagcg ccatgaagct
     3481 gttgtccgac ggacgatgga aaggagccag aagccaaggc agaagtctaa ccgctggtcc
     3541 tggggaagcc ctctccatgg gagctcgagc atccacagtg gagatccaga caggcgctca
     3601 gtttccacca tgaatctttc gaaacatgtt gatcctgtca ttagcaagcg gctctcctcc
     3661 tcgtctgcaa ctttgctaaa ctctccagat agagctcgcc gcctgcagct cagcccctgg
     3721 gagagcagcg ttgttagcag acttctgacg cccacacatt cgttcctggc cagaagcaaa
     3781 agcacggccg ccttgtctgg agacacagtt atccccattt gtcctcgttc agcatcttgc
     3841 agccccatca tcatgccctt caaagctgca cactctagaa acccagtgga ccgaccaaaa
     3901 ctctttgtaa caccgcctga gggctctgca cgaaggagga ccattcatgg actagcgagc
     3961 cataaaagag agcgagaaag agaacacgtt cccttccacg tttccccggg atccaccgga
     4021 tctagataac tgatcataat cagccatacc acatttggta gaggttttac ttgctttaaa
     4081 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     4141 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4201 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4261 acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     4321 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     4381 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     4441 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     4501 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     4561 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     4621 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     4681 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     4741 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4801 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     4861 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     4921 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     4981 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     5041 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     5101 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     5161 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     5221 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     5281 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     5341 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     5401 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     5461 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     5521 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     5581 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     5641 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     5701 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     5761 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     5821 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     5881 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     5941 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     6001 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     6061 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     6121 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     6181 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     6241 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     6301 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     6361 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     6421 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     6481 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     6541 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     6601 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6661 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6721 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6781 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6841 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6901 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6961 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     7021 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     7081 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     7141 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     7201 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     7261 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     7321 ttctgtggat aaccgtatta ccgccatgca t
//
