LOCUS       sequence-64891-e        6121 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 6121)
  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 6121)
  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-Start-1351
FEATURES             Location/Qualifiers
     source          1..6121
                     /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    2909..3030
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2946..2965)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3000..3019
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3037..3492)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3174..3195)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3386..3405
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3519..3623
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3625..3982
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3625..3816
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3648..3668)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3833..3968
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4017..4811
                     /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             4017..4811
                     /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(4071..4090)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4681..4700
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4999..5018)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5043..5090
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5419..6007
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5908..5927
                     /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 attctgcagt
     2761 cgacggtacc gcgggcccgg gatccaccgg atctagataa ctgatcataa tcagccatac
     2821 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     2881 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     2941 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     3001 tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt
     3061 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     3121 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     3181 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     3241 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     3301 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     3361 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     3421 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     3481 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3541 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3601 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     3661 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3721 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3781 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3841 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3901 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     3961 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     4021 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     4081 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     4141 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     4201 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     4261 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4321 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     4381 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     4441 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4501 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     4561 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4621 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4681 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4741 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4801 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     4861 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4921 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4981 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     5041 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     5101 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     5161 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     5221 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     5281 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5341 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5401 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5461 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5521 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     5581 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5641 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5701 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5761 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     5821 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     5881 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5941 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6001 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     6061 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
     6121 t
//
