LOCUS       sequence-246261-        7375 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SUN1_888 tagged with GFP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7375)
  AUTHORS   Nishioka Y, Imaizumi H, Imada J, Katahira J, Matsuura N, Hieda M
  TITLE     SUN1 splice variants, SUN1_888, SUN1_785, and predominant SUN1_916, 
            variably function in directional cell migration.
  JOURNAL   Nucleus. 2016 Nov;7(6):572-584. doi: 10.1080/19491034.2016.1260802.
  PUBMED    27858498
REFERENCE   2  (bases 1 to 7375)
  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: 
            "10.1080/19491034.2016"; journalName: "Nucleus"; date: "2016-11"; 
            volume: "7"; issue: "6"; pages: "572-584"
COMMENT     Sequence Label: pEGFP-SUN1_888
FEATURES             Location/Qualifiers
     source          1..7375
                     /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      615..624
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3315..3324
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3321..3323,3324..3326,3327..4040)
                     /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(3321..3323,3324..3326,3327..4037)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3366..3387)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3627..3646)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3974..3995
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4163..4284
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4200..4219)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4254..4273
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4291..4746)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4428..4449)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4640..4659
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4773..4877
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4879..5236
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4879..5070
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4902..4922)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5087..5222
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5271..6065
                     /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             5271..6065
                     /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(5325..5344)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5935..5954
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6253..6272)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6297..6344
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6673..7261
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7162..7181
                     /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 ccggactcag atctgccacc atggattttt ctcggcttca catgtacagt cctccccagt
      661 gtgtgccgga gaacacgggc tacacgtatg cgctcagttc cagctattct tcagatgctc
      721 tggattttga gacggagcac aaattggacc ctgtatttga ttctccacgg atgtcccgcc
      781 gtagtttgcg cctggccacg acagcatgca ccctggggga tggtgaggct gtgggtgccg
      841 acagcggcac cagcagcgct gtctccctga agaaccgagc ggccagaaca acaaaacagc
      901 gcagaagcac aaacaaatca gcttttagta tcaaccacgt gtcaaggcag gtcacgtcct
      961 ctggcgtcag ccacggcggc actgtcagcc tgcaggatgc tgtgactcga cggcctcctg
     1021 tattggacga gtcttggatt cgtgaacaga ccacagtgga ccacttctgg ggtcttgatg
     1081 atgatggtga tcttaaaggt ggaaataaag ctgccattca gggaaacggg gatgtgggag
     1141 ccgccgccgc caccgcgcac aacggcttct cctgcagcaa ctgcagcatg ctgtccgagc
     1201 gcaaggacgt gctcacggcg caccccgcgg cccccgggcc cgtgtcgaga gtttattcta
     1261 gggacaggaa tcaaaaatgc ggtgcgtctt tctacgtgaa taggattttg tggctggcca
     1321 gatacactgc atcatctttt tcatcatttt tagttcaact ttttcaagtg gttttaatga
     1381 agctcagtta tgaatcagaa aattacaaat tgaaaactca tgaatcaaaa gattgtgaat
     1441 ccgaaagcta taagtcaaaa agccatgaat caaaaggcga cgactgtaag ggcaagaggc
     1501 acctcgacgc gcacacagcc gcccactcgc agtcgccacg gctgcccggt cgggcaggga
     1561 ccctctggca catctgggca tgtgcaggtt acttcttgct gcagattctg cgcaggatcg
     1621 gagctgtggg ccaggctgtg tccaggacgg cgtggtcggc cctttggctg gccgtggttg
     1681 ctccagggaa ggcagcctct ggagtgttct ggtggctggg gattggatgg taccagtttg
     1741 ttactttgat ttcttggctg aatgtgtttc ttcttaccag gtgccttcga aacatctgca
     1801 agtttttagt cttgctcatc ccactcttcc ttttactagc aggtctctcc ttacggggcc
     1861 agggcaattt cttttcgttc ttgcccgtgt tgaactgggc aagcatgcat agaacacagc
     1921 gggtggatga cccccaggac gtgtttaaac ccacgacttc tcgcctgaag cagcctctgc
     1981 agggtgacag tgaggctttt ccgtggcatt ggatgagtgg cgtggagcag caggtggcct
     2041 ctctgtctgg acagtgccac caccatggtg agaatctccg agagctgacc actttgctac
     2101 agaagctgca ggctcgggtg gaccagatgg aaggcggcgc tgccgggccg tcagcttcgg
     2161 tcagagacgc tgtgggacag cccccgaggg agactgactt tatggccttt caccaagaac
     2221 atgaagtgcg tatgtcacac ttggaagata ttctgggaaa actgagagaa aaatctgagg
     2281 ccatccagaa ggaactagaa cagaccaagc aaaaaacaat cagtgcggtt ggtgagcagc
     2341 tcctgcccac ggtcgagcac ctccagctgg agctggatca gctaaagtca gagctgtcca
     2401 gctggcgaca cgtgaagacc ggctgtgaga cagtggatgc cgtacaagaa agagtggacg
     2461 tgcaagtcag agaaatggtg aaactcctgt tttccgaaga tcagcaaggc ggttctctgg
     2521 aacagctgct gcagaggttc tcatcacagt ttgtgagcaa aggcgacttg cagacgatgc
     2581 tgcgagacct gcagctgcag atcctgcgga acgtcaccca ccacgtttcc gtgaccaagc
     2641 agctcccaac ctcagaagcc gtggtgtctg ctgtgagcga ggcgggggcg tctggaataa
     2701 cagaggcgca agcacgtgcc atcgtgaaca gcgccttgaa gctgtattcc caagataaga
     2761 ccgggatggt ggactttgct ctggaatctg gtggtggcag catcttgagt actcgctgtt
     2821 ctgaaactta cgaaaccaaa acggcgctga tgagtctgtt tgggatcccg ctgtggtact
     2881 tctcgcagtc cccgcgcgtg gtcatccagc ctgacattta ccccggtaac tgctgggcat
     2941 ttaaaggctc ccaggggtac ctggtggtga ggctctccat gatgatccac ccagccgcct
     3001 tcactctgga gcacatccct aagacgctgt cgccaacagg caacatcagc agcgccccca
     3061 aggacttcgc cgtctatgga ttagaaaatg agtatcagga agaagggcag cttctgggac
     3121 agttcacgta tgatcaggat ggggagtcgc tccagatgtt ccaggccctg aaaagacccg
     3181 acgacacagc tttccaaata gtggaacttc ggattttttc taactggggc catcctgagt
     3241 atacctgtct gtatcggttc agagttcatg gcgaacctgt caaggtcgac ggtaccgcgg
     3301 gcccgggatc catcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     3361 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     3421 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     3481 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     3541 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     3601 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     3661 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     3721 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     3781 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     3841 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     3901 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     3961 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     4021 tggacgagct gtacaagtaa agcggccgcg actctagatc ataatcagcc ataccacatt
     4081 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     4141 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     4201 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     4261 gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat
     4321 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     4381 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     4441 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     4501 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     4561 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     4621 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     4681 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     4741 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4801 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4861 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     4921 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4981 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     5041 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     5101 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     5161 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     5221 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     5281 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     5341 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     5401 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     5461 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     5521 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     5581 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     5641 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     5701 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     5761 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     5821 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     5881 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     5941 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     6001 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     6061 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     6121 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     6181 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     6241 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     6301 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     6361 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     6421 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     6481 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     6541 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     6601 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     6661 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6721 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     6781 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     6841 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     6901 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6961 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     7021 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     7081 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     7141 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     7201 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     7261 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     7321 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcat
//
