LOCUS       sequence-433864-        7457 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SUN1_916 tagged with GFP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7457)
  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 7457)
  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_916
FEATURES             Location/Qualifiers
     source          1..7457
                     /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      3397..3406
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3403..3405,3406..3408,3409..4122)
                     /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(3403..3405,3406..3408,3409..4119)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3448..3469)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3709..3728)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4056..4077
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4245..4366
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4282..4301)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4336..4355
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4373..4828)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4510..4531)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4722..4741
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4855..4959
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4961..5318
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4961..5152
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4984..5004)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5169..5304
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5353..6147
                     /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             5353..6147
                     /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(5407..5426)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6017..6036
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6335..6354)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6379..6426
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6755..7343
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7244..7263
                     /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 atctgcccat ggatttttct cggcttcaca tgtacagtcc tccccagtgt
      661 gtgccggaga acacgggcta cacgtatgcg ctcagttcca gctattcttc agatgctctg
      721 gattttgaga cggagcacaa attggaccct gtatttgatt ctccacggat gtcccgccgt
      781 agtttgcgcc tggccacgac agcatgcacc ctgggggatg gtgaggctgt gggtgccgac
      841 agcggcacca gcagcgctgt ctccctgaag aaccgagcgg ccagaacaac aaaacagcgc
      901 agaagcacaa acaaatcagc ttttagtatc aaccacgtgt caaggcaggt cacgtcctct
      961 ggcgtcagcc acggcggcac tgtcagcctg caggatgctg tgactcgacg gcctcctgta
     1021 ttggacgagt cttggattcg tgaacagacc acagtggacc acttctgggg tcttgatgat
     1081 gatggtgatc ttaaaggtgg aaataaagct gccattcagg gaaacgggga tgtgggagcc
     1141 gccgccgcca ccgcgcacaa cggcttctcc tgcagcaact gcagcatgct gtccgagcgc
     1201 aaggacgtgc tcacggcgca ccccgcggcc cccgggcccg tgtcgagagt ttattctagg
     1261 gacaggaatc aaaaatgcgg tgcgtctttc tacgtgaata ggattttgtg gctggccaga
     1321 tacactgcat catctttttc atcattttta gttcaacttt ttcaagtggt tttaatgaag
     1381 ctcagttatg aatcagaaaa ttacaaattg aaaactcatg aatcaaaaga ttgtgaatcc
     1441 gaaagctata agtcaaaaag ccatgaatca aaagctcatg ccagttacta tgggagaatg
     1501 aatgtgagag aggttctcag agaggatggc cacctcagtg taaatgggga agcgctgtgc
     1561 gacgactgta agggcaagag gcacctcgac gcgcacacag ccgcccactc gcagtcgcca
     1621 cggctgcccg gtcgggcagg gaccctctgg cacatctggg catgtgcagg ttacttcttg
     1681 ctgcagattc tgcgcaggat cggagctgtg ggccaggctg tgtccaggac ggcgtggtcg
     1741 gccctttggc tggccgtggt tgctccaggg aaggcagcct ctggagtgtt ctggtggctg
     1801 gggattggat ggtaccagtt tgttactttg atttcttggc tgaatgtgtt tcttcttacc
     1861 aggtgccttc gaaacatctg caagttttta gtcttgctca tcccactctt ccttttacta
     1921 gcaggtctct ccttacgggg ccagggcaat ttcttttcgt tcttgcccgt gttgaactgg
     1981 gcaagcatgc atagaacaca gcgggtggat gacccccagg acgtgtttaa acccacgact
     2041 tctcgcctga agcagcctct gcagggtgac agtgaggctt ttccgtggca ttggatgagt
     2101 ggcgtggagc agcaggtggc ctctctgtct ggacagtgcc accaccatgg tgagaatctc
     2161 cgagagctga ccactttgct acagaagctg caggctcggg tggaccagat ggaaggcggc
     2221 gctgccgggc cgtcagcttc ggtcagagac gctgtgggac agcccccgag ggagactgac
     2281 tttatggcct ttcaccaaga acatgaagtg cgtatgtcac acttggaaga tattctggga
     2341 aaactgagag aaaaatctga ggccatccag aaggaactag aacagaccaa gcaaaaaaca
     2401 atcagtgcgg ttggtgagca gctcctgccc acagtcgagc acctccagct ggagctggat
     2461 cagctaaagt cagagctgtc cagctggcga cacgtgaaga ccggctgtga gacagtggat
     2521 gccgtacaag aaagagtgga cgtgcaagtc agagaaatgg tgaaactcct gttttccgaa
     2581 gatcagcaag gcggttctct ggaacagctg ctgcagaggt tctcatcaca gtttgtgagc
     2641 aaaggcgact tgcagacgat gctgcgagac ctgcagctgc agatcctgcg gaacgtcacc
     2701 caccacgttt ccgtgaccaa gcagctccca acctcagaag ccgtggtgtc tgctgtgagc
     2761 gaggcggggg cgtctggaat aacagaggcg caagcacgtg ccatcgtgaa cagcgccttg
     2821 aagctgtatt cccaagataa gaccgggatg gtggactttg ctctggaatc tggtggtggc
     2881 agcatcttga gtactcgctg ttctgaaact tacgaaacca aaacggcgct gatgagtctg
     2941 tttgggatcc cgctgtggta cttctcgcag tccccgcgcg tggtcatcca gcctgacatt
     3001 taccccggta actgctgggc atttaaaggc tcccaggggt acctggtggt gaggctctcc
     3061 atgatgatcc acccagccgc cttcactctg gagcacatcc ctaagacgct gtcgccaaca
     3121 ggcaacatca gcagcgcccc caaggacttc gccgtctatg gattagaaaa tgagtatcag
     3181 gaagaagggc agcttctggg acagttcacg tatgatcagg atggggagtc gctccagatg
     3241 ttccaggccc tgaaaagacc cgacgacaca gctttccaaa tagtggaact tcggattttt
     3301 tctaactggg gccatcctga gtatacctgt ctgtatcggt tcagagttca tggcgaacct
     3361 gtcaaggtcg acggtaccgc gggcccggga tccatcgcca ccatggtgag caagggcgag
     3421 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     3481 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     3541 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     3601 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     3661 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     3721 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     3781 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     3841 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     3901 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     3961 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     4021 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     4081 gccgccggga tcactctcgg catggacgag ctgtacaagt aaagcggccg cgactctaga
     4141 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     4201 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     4261 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     4321 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta
     4381 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     4441 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     4501 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     4561 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     4621 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     4681 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     4741 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     4801 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4861 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4921 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     4981 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     5041 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     5101 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     5161 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     5221 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     5281 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     5341 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     5401 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     5461 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     5521 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     5581 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     5641 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     5701 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     5761 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     5821 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     5881 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     5941 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     6001 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     6061 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     6121 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     6181 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     6241 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     6301 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     6361 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     6421 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     6481 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     6541 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     6601 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6661 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6721 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6781 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6841 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6901 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6961 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     7021 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     7081 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     7141 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     7201 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     7261 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     7321 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     7381 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     7441 gtattaccgc catgcat
//
