LOCUS       sequence-246266-        7066 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SUN1_785 tagged with GFP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7066)
  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 7066)
  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_785
FEATURES             Location/Qualifiers
     source          1..7066
                     /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      3006..3015
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3012..3014,3015..3017,3018..3731)
                     /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(3012..3014,3015..3017,3018..3728)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3057..3078)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3318..3337)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3665..3686
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3854..3975
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3891..3910)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3945..3964
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3982..4437)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4119..4140)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4331..4350
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4464..4568
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4570..4927
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4570..4761
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4593..4613)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4778..4913
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4962..5756
                     /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             4962..5756
                     /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(5016..5035)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5626..5645
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5944..5963)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5988..6035
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6364..6952
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6853..6872
                     /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 tcaaaaatgt tacttcttgc tgcagattct gcgcaggatc ggagctgtgg
     1321 gccaggctgt gtccaggacg gcgtggtcgg ccctttggct ggccgtggtt gctccaggga
     1381 aggcagcctc tggagtgttc tggtggctgg ggattggatg gtaccagttt gttactttga
     1441 tttcttggct gaatgtgttt cttcttacca ggtgccttcg aaacatctgc aagtttttag
     1501 tcttgctcat cccactcttc cttttactag caggtctctc cttacggggc cagggcaatt
     1561 tcttttcgtt cttgcccgtg ttgaactggg caagcatgca tagaacacag cgggtggatg
     1621 acccccagga cgtgtttaaa cccacgactt ctcgcctgaa gcagcctctg cagggtgaca
     1681 gtgaggcttt tccgtggcat tggatgagtg gcgtggagca gcaggtggcc tctctgtctg
     1741 gacagtgcca ccaccatggt gagaatctcc gagagctgac cactttgcta cagaagctgc
     1801 aggctcgggt ggaccagatg gaaggcggcg ctgccgggcc gtcagcttcg gtcagagacg
     1861 ctgtgggaca gcccccgagg gagactgact ttatggcctt tcaccaagaa catgaagtgc
     1921 gtatgtcaca cttggaagat attctgggaa aactgagaga aaaatctgag gccatccaga
     1981 aggaactaga acagaccaag caaaaaacaa tcagtgcggt tggtgagcag ctcctgccca
     2041 cggtcgagca cctccagctg gagctggatc agctaaagtc agagctgtcc agctggcgac
     2101 acgtgaagac cggctgtgag acagtggatg ccgtacaaga aagagtggac gtgcaagtca
     2161 gagaaatggt gaaactcctg ttttccgaag atcagcaagg cggttctctg gaacagctgc
     2221 tgcagaggtt ctcatcacag tttgtgagca aaggcgactt gcagacgatg ctgcgagacc
     2281 tgcagctgca gatcctgcgg aacgtcaccc accacgtttc cgtgaccaag cagctcccaa
     2341 cctcagaagc cgtggtgtct gctgtgagcg aggcgggggc gtctggaata acagaggcgc
     2401 aagcacgtgc catcgtgaac agcgccttga agctgtattc ccaagataag accgggatgg
     2461 tggactttgc tctggaatct ggtggtggca gcatcttgag tactcgctgt tctgaaactt
     2521 acgaaaccaa aacggcgctg atgagtctgt ttgggatccc gctgtggtac ttctcgcagt
     2581 ccccgcgcgt ggtcatccag cctgacattt accccggtaa ctgctgggca tttaaaggct
     2641 cccaggggta cctggtggtg aggctctcca tgatgatcca cccagccgcc ttcactctgg
     2701 agcacatccc taagacgctg tcgccaacag gcaacatcag cagcgccccc aaggacttcg
     2761 ccgtctatgg attagaaaat gagtatcagg aagaagggca gcttctggga cagttcacgt
     2821 atgatcagga tggggagtcg ctccagatgt tccaggccct gaaaagaccc gacgacacag
     2881 ctttccaaat agtggaactt cggatttttt ctaactgggg ccatcctgag tatacctgtc
     2941 tgtatcggtt cagagttcat ggcgaacctg tcaaggtcga cggtaccgcg ggcccgggat
     3001 ccatcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     3061 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     3121 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     3181 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     3241 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     3301 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     3361 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     3421 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     3481 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     3541 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     3601 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     3661 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     3721 tgtacaagta aagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt
     3781 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     3841 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     3901 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     3961 catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     4021 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     4081 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     4141 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     4201 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     4261 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     4321 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     4381 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     4441 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4501 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4561 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     4621 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4681 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4741 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     4801 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     4861 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     4921 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     4981 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     5041 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     5101 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     5161 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     5221 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     5281 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     5341 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     5401 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     5461 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     5521 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     5581 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     5641 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     5701 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     5761 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     5821 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     5881 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     5941 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     6001 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     6061 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     6121 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     6181 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     6241 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     6301 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     6361 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6421 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     6481 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     6541 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6601 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6661 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6721 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6781 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6841 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6901 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6961 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     7021 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
