LOCUS       sequence-178207-        5189 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human RBM3 coupled to GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5189)
  AUTHORS   Rossello CA, Lindstrom L, Glindre J, Eklund G, Alvarado-Kristensson 
            M
  TITLE     Gamma-tubulin coordinates nuclear envelope assembly around 
            chromatin.
  JOURNAL   Heliyon. 2016 Sep 24;2(9):e00166. eCollection 2016 Sep.
  PUBMED    27699285
REFERENCE   2  (bases 1 to 5189)
  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"; journalName: "Heliyon."; 
            date: "2016-09-24"; volume: "2(9)"; pages: "e00166. eCollection 2016
            Sep"
COMMENT     Sequence Label: pEGFPN1-RBM3
FEATURES             Location/Qualifiers
     source          1..5189
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      97..232
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             281..1075
                     /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             281..1075
                     /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(335..354)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     945..964
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1263..1282)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1307..1354
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1683..2271
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2172..2191
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2446..2749
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2750..2953
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2904..2924
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3514..3523
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3520..3522,3523..3525,3526..4239)
                     /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(3520..3522,3523..3525,3526..4236)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3565..3586)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3826..3845)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4173..4194
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4362..4483
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4399..4418)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4453..4472
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4490..4945)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4627..4648)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4839..4858
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4972..5076
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(5078..5189,1..246)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        join(5078..5189,1..80)
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5101..5121)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
       61 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
      121 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
      181 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
      241 tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
      301 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
      361 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
      421 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct
      481 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
      541 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
      601 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
      661 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
      721 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
      781 agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac
      841 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
      901 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
      961 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     1021 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     1081 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     1141 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     1201 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta
     1261 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     1321 cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     1381 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     1441 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     1501 gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt
     1561 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     1621 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     1681 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     1741 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     1801 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     1861 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     1921 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     1981 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     2041 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     2101 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     2161 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     2221 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     2281 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     2341 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta
     2401 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     2461 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     2521 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     2581 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     2641 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     2701 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     2761 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     2821 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     2881 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     2941 tataagcaga gctggtttag tgaaccgtca gatccgctag cgaccatgtc ctctgaagaa
     3001 ggaaagctct tcgtgggagg gctcaacttt aacaccgacg agcaggcact ggaagaccac
     3061 ttcagcagtt tcggacctat ctctgaggtg gtcgttgtca aggaccggga gactcagcgg
     3121 tccaggggtt ttggtttcat caccttcacc aacccagagc atgcttcagt tgccatgaga
     3181 gccatgaacg gagagtctct ggatggtcgt cagatccgtg tggatcatgc aggcaagtct
     3241 gctcggggaa ccagaggagg tggctttggg gcccatgggc gtggtcgcag ctactctaga
     3301 ggtggtgggg accagggcta tgggagtggc aggtattatg acagtcgacc tggagggtat
     3361 ggatatggat atggacgttc cagagactat aatggcagaa accagggtgg ttatgaccgc
     3421 tactcaggag gaaattacag agacaattat gacaacatga aaaagcttcg aattctgcag
     3481 tcgacggtac cgcgggcccg ggatccaccg gtcgccacca tggtgagcaa gggcgaggag
     3541 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     3601 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     3661 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     3721 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     3781 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     3841 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     3901 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     3961 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     4021 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     4081 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     4141 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     4201 gccgggatca ctctcggcat ggacgagctg tacaagtaaa gcggccgcga ctctagatca
     4261 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     4321 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     4381 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     4441 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc
     4501 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     4561 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     4621 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     4681 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     4741 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     4801 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     4861 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     4921 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     4981 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5041 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt
     5101 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     5161 aaagcatgca tctcaattag tcagcaacc
//
