LOCUS       sequence-482056-        8233 bp    DNA     circular SYN 05-AUG-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 8233)
  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 8233)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 5, 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..8233
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        764..1067
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1068..1271
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1222..1242
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     mobile_element  2012..2779
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     regulatory      4876..4885
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4882..4884,4885..4887,4888..5601)
                     /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(4882..4884,4885..4887,4888..5598)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4927..4948)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5188..5207)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5535..5556
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5724..5845
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5761..5780)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5815..5834
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5852..6307)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5989..6010)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6201..6220
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6334..6438
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6440..6797
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6440..6631
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6463..6483)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6648..6783
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6832..7626
                     /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             6832..7626
                     /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(6886..6905)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7496..7515
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7814..7833)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7858..7905
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
      721 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      781 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      841 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      901 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      961 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     1021 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     1081 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     1141 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     1201 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     1261 taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggact cagatctgcc
     1321 catggatttt tctcggcttc acatgtacag tcctccccag tgtgtgccgg agaacacggg
     1381 ctacacgtat gcgctcagtt ccagctattc ttcagatgct ctggattttg agacggagca
     1441 caaattggac cctgtatttg attctccacg gatgtcccgc cgtagtttgc gcctggccac
     1501 gacagcatgc accctggggg atggtgaggc tgtgggtgcc gacagcggca ccagcagcgc
     1561 tgtctccctg aagaaccgag cggccagaac aacaaaacag cgcagaagca caaacaaatc
     1621 agcttttagt atcaaccacg tgtcaaggca ggtcacgtcc tctggcgtca gccacggcgg
     1681 cactgtcagc ctgcaggatg ctgtgactcg acggcctcct gtattggacg agtcttggat
     1741 tcgtgaacag accacagtgg accacttctg gggtcttgat gatgatggtg atcttaaagg
     1801 tggaaataaa gctgccattc agggaaacgg ggatgtggga gccgccgccg ccaccgcgca
     1861 caacggcttc tcctgcagca actgcagcat gctgtccgag cgcaaggacg tgctcacggc
     1921 gcaccccgcg gcccccgggc ccgtgtcgag agtttattct agggacagga atcaaaaatg
     1981 cggtgcgtct ttctacgtga ataggatttt gggtgatgct gccaacttac tgatttagtg
     2041 tatgatggtg tttttgaggt gctccagtgg cttctgtttc tatcagctgt ccctcctgtt
     2101 cagctactga cggggtggtg cgtaacggca aaagcaccgc cggacatcag cgctatctct
     2161 gctctcactg ccgtaaaaca tggcaactgc agttcactta caccgcttct caacccggta
     2221 cgcaccagaa aatcattgat atggccatga atggcgttgg atgccgggca acagcccgca
     2281 ttatgggcgt tggcctcaac acgattttac gtcacttaaa aaactcaggc cgcagtcggt
     2341 aacctcgcgc atacagccgg gcagtgacgt catcgtctgc gcggaaatgg acgaacagtg
     2401 gggctatgtc ggggctaaat cgcgccagcg ctggctgttt tacgcgtatg acagtctccg
     2461 gaagacggtt gttgcgcacg tattcggtga acgcactatg gcgacgctgg ggcgtcttat
     2521 gagcctgctg tcaccctttg acgtggtgat atggatgacg gatggctggc cgctgtatga
     2581 atcccgcctg aagggaaagc tgcacgtaat cagcaagcga tatacgcagc gaattgagcg
     2641 gcataacctg aatctgaggc agcacctggc acggctggga cggaagtcgc tgtcgttctc
     2701 aaaatcggtg gagctgcatg acaaagtcat cgggcattat ctgaacataa aacactatca
     2761 ataagttgga gtcattaccg gattttgtgg ctggccagat acactgcatc atctttttca
     2821 tcatttttag ttcaactttt tcaagtggtt ttaatgaagc tcagttatga atcagaaaat
     2881 tacaaattga aaactcatga atcaaaagat tgtgaatccg aaagctataa gtcaaaaagc
     2941 catgaatcaa aagctcatgc cagttactat gggagaatga atgtgagaga ggttctcaga
     3001 gaggatggcc acctcagtgt aaatggggaa gcgctgtgcg acgactgtaa gggcaagagg
     3061 cacctcgacg cgcacacagc cgcccactcg cagtcgccac ggctgcccgg tcgggcaggg
     3121 accctctggc acatctgggc atgtgcaggt tacttcttgc tgcagattct gcgcaggatc
     3181 ggagctgtgg gccaggctgt gtccaggacg gcgtggtcgg ccctttggct ggccgtggtt
     3241 gctccaggga aggcagcctc tggagtgttc tggtggctgg ggattggatg gtaccagttt
     3301 gttactttga tttcttggct gaatgtgttt cttcttacca ggtgccttcg aaacatctgc
     3361 aagtttttag tcttgctcat cccactcttc cttttactag caggtctctc cttacggggc
     3421 cagggcaatt tcttttcgtt cttgcccgtg ttgaactggg caagcatgca tagaacacag
     3481 cgggtggatg acccccagga cgtgtttaaa cccacgactt ctcgcctgaa gcagcctctg
     3541 cagggtgaca gtgaggcttt tccgtggcat tggatgagtg gcgtggagca gcaggtggcc
     3601 tctctgtctg gacagtgcca ccaccatggt gagaatctcc gagagctgac cactttgcta
     3661 cagaagctgc aggctcgggt ggaccagatg gaaggcggcg ctgccgggcc gtcagcttcg
     3721 gtcagagacg ctgtgggaca gcccccgagg gagactgact ttatggcctt tcaccaagaa
     3781 catgaagtgc gtatgtcaca cttggaagat attctgggaa aactgagaga aaaatctgag
     3841 gccatccaga aggaactaga acagaccaag caaaaaacaa tcagtgcggt tggtgagcag
     3901 ctcctgccca cagtcgagca cctccagctg gagctggatc agctaaagtc agagctgtcc
     3961 agctggcgac acgtgaagac cggctgtgag acagtggatg ccgtacaaga aagagtggac
     4021 gtgcaagtca gagaaatggt gaaactcctg ttttccgaag atcagcaagg cggttctctg
     4081 gaacagctgc tgcagaggtt ctcatcacag tttgtgagca aaggcgactt gcagacgatg
     4141 ctgcgagacc tgcagctgca gatcctgcgg aacgtcaccc accacgtttc cgtgaccaag
     4201 cagctcccaa cctcagaagc cgtggtgtct gctgtgagcg aggcgggggc gtctggaata
     4261 acagaggcgc aagcacgtgc catcgtgaac agcgccttga agctgtattc ccaagataag
     4321 accgggatgg tggactttgc tctggaatct ggtggtggca gcatcttgag tactcgctgt
     4381 tctgaaactt acgaaaccaa aacggcgctg atgagtctgt ttgggatccc gctgtggtac
     4441 ttctcgcagt ccccgcgcgt ggtcatccag cctgacattt accccggtaa ctgctgggca
     4501 tttaaaggct cccaggggta cctggtggtg aggctctcca tgatgatcca cccagccgcc
     4561 ttcactctgg agcacatccc taagacgctg tcgccaacag gcaacatcag cagcgccccc
     4621 aaggacttcg ccgtctatgg attagaaaat gagtatcagg aagaagggca gcttctggga
     4681 cagttcacgt atgatcagga tggggagtcg ctccagatgt tccaggccct gaaaagaccc
     4741 gacgacacag ctttccaaat agtggaactt cggatttttt ctaactgggg ccatcctgag
     4801 tatacctgtc tgtatcggtt cagagttcat ggcgaacctg tcaaggtcga cggtaccgcg
     4861 ggcccgggat ccatcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
     4921 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     4981 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     5041 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     5101 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     5161 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     5221 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     5281 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     5341 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     5401 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     5461 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     5521 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     5581 atggacgagc tgtacaagta aagcggccgc gactctagat cataatcagc cataccacat
     5641 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     5701 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     5761 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     5821 tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa
     5881 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     5941 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     6001 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     6061 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     6121 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     6181 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     6241 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     6301 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     6361 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     6421 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     6481 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     6541 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     6601 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     6661 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     6721 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     6781 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     6841 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     6901 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     6961 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     7021 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     7081 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     7141 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     7201 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     7261 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     7321 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     7381 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     7441 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     7501 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     7561 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     7621 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     7681 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     7741 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     7801 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     7861 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     7921 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     7981 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     8041 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     8101 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     8161 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     8221 tcaaaggatc ttc
//
