LOCUS       sequence-418287-        5411 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Tetraspanin-4 (TSPAN4) as migrasome marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5411)
  AUTHORS   Ma L, Li Y, Peng J, Wu D, Zhao X, Cui Y, Chen L, Yan X, Du Y, Yu L
  TITLE     Discovery of the migrasome, an organelle mediating release of 
            cytoplasmic contents during cell migration.
  JOURNAL   Cell Res. 2015 Jan;25(1):24-38. doi: 10.1038/cr.2014.135. Epub 2014 
            Oct 24.
  PUBMED    25342562
REFERENCE   2  (bases 1 to 5411)
  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: 
            "10.1038/cr.2014.135"; journalName: "Cell Res"; date: "2015-01"; 
            volume: "25"; issue: "1"; pages: "24-38"
COMMENT     Sequence Label: pEGFP-N1-Tspan4
FEATURES             Location/Qualifiers
     source          1..5411
                     /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      1351..1360
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1357..1359,1360..1362,1363..2076)
                     /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(1357..1359,1360..1362,1363..2073)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1402..1423)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1663..1682)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2010..2031
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2199..2320
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2236..2255)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2290..2309
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2327..2782)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2464..2485)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2676..2695
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2809..2913
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2915..3272
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2915..3106
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2938..2958)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3123..3258
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3307..4101
                     /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             3307..4101
                     /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(3361..3380)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3971..3990
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4289..4308)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4333..4380
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4709..5297
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5198..5217
                     /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 atctcgagac catggcgcgc ggctgcctcc agggcgtcaa gtacctcatg
      661 ttcgccttca acctgctctt ctggctgggt ggctgtggtg tcctgggtgt tggcatctgg
      721 ttggctgcca cacagggaaa ctttgccacc ttatcatcct catttccatc cttgtcggct
      781 gccaacctgc tcatcgtcac cgggaccttc gtcatggcca tcggcttcgt gggctgcatt
      841 ggggccctca aggagaacaa gtgcctactg ctcactttct ttgtgctgct gctgctagtg
      901 ttcctgctgg aagccaccat tgctgtgctc ttctttgcct acagtgacaa gattgacagt
      961 tatgcccaac aagacctgaa gaagggcctg catctgtatg gcacacaggg caacgtgggc
     1021 ctcaccaatg cctggagcat catccagact gatttccgat gctgtggagt ttccaattac
     1081 actgattggt ttgaggtata caatgccact cgtgtgcctg actcctgctg tctggagttc
     1141 agtgatagct gtgggttaca tgaacctggt acctggtgga agtcgccctg ttatgagaca
     1201 gtgaaggcct ggctccagga gaacctgcta gctgtgggca tctttggact gtgcacggca
     1261 ctggtgcaga ttctgggcct caccttcgct atgaccatgt actgccaggt ggtaaaggcg
     1321 gacacctact gtgcacggga tccaccggtc gccaccatgg tgagcaaggg cgaggagctg
     1381 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     1441 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     1501 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     1561 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     1621 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     1681 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     1741 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     1801 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     1861 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     1921 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     1981 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     2041 gggatcactc tcggcatgga cgagctgtac aagtaaagcg gccgcgactc tagatcataa
     2101 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     2161 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     2221 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     2281 attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt
     2341 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     2401 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     2461 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     2521 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     2581 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     2641 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     2701 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     2761 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     2821 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2881 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     2941 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3001 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3061 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3121 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3181 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     3241 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     3301 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3361 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3421 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     3481 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     3541 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     3601 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     3661 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     3721 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     3781 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     3841 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     3901 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     3961 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4021 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4081 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     4141 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     4201 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     4261 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     4321 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     4381 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     4441 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     4501 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     4561 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4621 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4681 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4741 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4801 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     4861 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4921 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4981 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5041 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5101 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5161 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5221 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5281 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5341 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     5401 ccgccatgca t
//
