LOCUS       sequence-419657-        7353 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminally truncated GFP-RepoMan.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7353)
  AUTHORS   Pfisterer M, Robert R, Saul VV, Pritz A, Seibert M, Feederle R, 
            Schmitz ML
  TITLE     The Aurora B-controlled PP1/RepoMan complex determines the spatial 
            and temporal distribution of mitotic H2B S6 phosphorylation.
  JOURNAL   Open Biol. 2024 May;14(5):230460. doi: 10.1098/rsob.230460. Epub 
            2024 May 29.
  PUBMED    38806145
REFERENCE   2  (bases 1 to 7353)
  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: "Open 
            Biol."; date: "2024-05"; volume: "14(5):230460. doi"; pages: " 
            10.1098/rsob.230460. Epub 2024 May 29"
COMMENT     Sequence Label: pEGFP-Repo-Man-1-890
FEATURES             Location/Qualifiers
     source          1..7353
                     /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      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4213..4334
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4250..4269)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4304..4323
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4341..4796)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4478..4499)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4690..4709
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4823..4927
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4952..4972)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4996..5206
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        5018..5214
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5065..5200
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5249..6043
                     /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             5249..6043
                     /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(5303..5322)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5913..5932
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6231..6250)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6275..6322
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6651..7239
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7140..7159
                     /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 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc
     1381 cgccggcgaa tggatgccaa ttcaaaagac aagccccctg aaaccaagga gtctgcaatg
     1441 aataatgctg gaaatgcctc tttcattttg ggaactggga agattgtgac tcctcagaag
     1501 catgccgaat tacctcctaa tccttgcaca ccagatactt ttaaatcacc tttgaacttt
     1561 tccacagtaa ccgtagagca attgggaatt acacctgaaa gctttgttag gaactctgca
     1621 ggaaagtcat catcctacct taaaaaatgt agacgacgtt ctgcagtcgg tgctcggggc
     1681 tctcctgaaa caaaccatct gattcgtttc attgctcggc agcaaaatat aaagaatgct
     1741 aggaaatctc ctttggcaca agattctcct tcccagggca gccctgcact gtatcgaaat
     1801 gttaacactt taagagaacg aatatcagcc ttccagtcag cttttcactc cataaaggaa
     1861 aacgagaaaa tgaccggctg tctggaattc tcagaggcag gaaaagagtc cgagatgacc
     1921 gatctgacaa ggaaggaagg tctcagcgct tgccagcagt ctgggttccc tgcagtgttg
     1981 tcctccaaac gtcggagaat atcctatcag agagactctg atgaaaatct gacggatgct
     2041 gaaggaaaag taattggtct ccagatattc aatattgata cagacagagc atgtgcagtt
     2101 gaaacttctg tcgaccttag cgaaatttca agcaagcttg gttcaacaca gtctggattt
     2161 ttagttgaag agtctcttcc cctttcagag ctcacagaga cttcaaatgc actaaaggtt
     2221 gctgactgtg tagtgggcaa aggatcaagt gatgccgttt cgcctgacac gttcacagca
     2281 gaagtgagct cagacgcagt ccctgatgtc aggtcaccag ctactccagc ctgcaggagg
     2341 gaccttccca cccccaagac ctttgtactt cgttctgtac tgaagaaacc ctctgttaag
     2401 atgtgtctag agagcttaca ggaacactgt aacaacctct atgatgatga tgggactcat
     2461 ccgagcttaa tctcaaatct cccaaactgt tgcaaagaga aagaagcaga agatgaagaa
     2521 aattttgaag cacctgcctt tctaaatatg aggaagagga agagagttac ttttggagag
     2581 gacttaagcc cggaagtgtt tgatgaatct ttgccagcaa atactccatt gcgtaaagga
     2641 ggaacacctg tttgtaaaaa agacttcagt ggtctcagtt ccctgctgct tgagcagtca
     2701 cctgttcctg agccattacc tcaaccagat tttgatgaca agggggagaa tcttgaaaac
     2761 atagaaccac ttcaagtatc atttgccgtt ctcagttctc ctaataaatc atcaatctct
     2821 gagacccttt caggcactga tacctttagt tcttcaaata accatgagaa aatatcctca
     2881 ccaaaggtgg gccgtatcac aaggacttct aaccggagga accagctggt tagcgtggta
     2941 gaagaaagcg tgtgtaatct cctgaacaca gaagttcagc cttgtaaaga aaagaaaatt
     3001 aataggagga agtctcaaga aacaaagtgt acaaagagag cacttcctaa gaagagtcag
     3061 gttttaaaaa gttgcagaaa gaagaaagga aagggaaaga aaagtgttca gaaatcttta
     3121 tatggggaaa gagacattgc ttctaagaag cccctcctca gtcctattcc cgagctgcct
     3181 gaagtccctg agatgacacc ttccattccg agcatccgaa gactgggttc aggttatttc
     3241 agttcaaatg gcaaactgga agaagtgaag actcctaaaa atccagtgaa aagaaaggat
     3301 cttttgcgtc atgacccaga tttgcatatg catcaaggct atgataaata tgatgtctct
     3361 gaattctgct cttatataaa aagttcctca tcgcttggca atgctacttc tgatgaggac
     3421 ccgaacacta acattatgaa tataaacgag aacaaaaata ttccaaaagc aaaaaataag
     3481 tcagaaagtg aaaatgaacc aaaagctgga actgacagtc ctgtttcttg tgcttctgta
     3541 actgaagaac gtgtggcatc agatagtccc aaacctgctc tgaccctgca gcagggtcaa
     3601 gaattttctg ctggtggtca aaatgcagaa aacctttgtc agttctttaa aatttcacca
     3661 gatttaaaca taaagtgtga aagaaaggat gacttcttag gagctgcaga aggaaaactg
     3721 caatgcaatc gtttaatgcc taattcacaa aaagactgtc attgtttagg agatgtctta
     3781 attgaaaata cgaaagaatc taaaagccag agtgaggatt tgggaagaaa acccatggaa
     3841 agtagcagtg ttgtgagttg cagagacagg aaagatagaa gacgttccat gtgttattct
     3901 gatggtcgaa gtttacattt ggaaaaaaat ggaaatcaca caccatcctc cagtgtgggc
     3961 agctctgtag aaattagttt agaaaattct gaactgttta aagatttgtc tgatgccatt
     4021 gagcaaacct ttcagaggag aaatagtgaa accaaagtga tgcccggggc tacgcttagc
     4081 ccgggatcca ccggatctag ataactgatc ataatcagcc ataccacatt tgtagaggtt
     4141 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     4201 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     4261 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     4321 atcaatgtat cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     4381 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     4441 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     4501 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     4561 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     4621 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     4681 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     4741 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg
     4801 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     4861 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     4921 gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     4981 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     5041 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     5101 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     5161 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
     5221 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     5281 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     5341 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     5401 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
     5461 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     5521 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     5581 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     5641 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     5701 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     5761 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     5821 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     5881 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     5941 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     6001 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     6061 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     6121 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     6181 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
     6241 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     6301 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     6361 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     6421 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     6481 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
     6541 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6601 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     6661 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     6721 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     6781 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     6841 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6901 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6961 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     7021 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     7081 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     7141 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     7201 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     7261 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     7321 gattctgtgg ataaccgtat taccgccatg cat
//
