LOCUS       sequence-422693-        6311 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human RAD18 tagged with EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6311)
  AUTHORS   Tsuji Y, Watanabe K, Araki K, Shinohara M, Yamagata Y, Tsurimoto T, 
            Hanaoka F, Yamamura K, Yamaizumi M, Tateishi S
  TITLE     Recognition of forked and single-stranded DNA structures by human 
            RAD18 complexed with RAD6B protein triggers its recruitment to 
            stalled replication forks.
  JOURNAL   Genes Cells. 2008 Apr;13(4):343-54. doi: 
            10.1111/j.1365-2443.2008.01176.x.
  PUBMED    18363965
REFERENCE   2  (bases 1 to 6311)
  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: 
            "10.1111/j.1365-2443.2008.01176"; journalName: "Genes Cells"; date: 
            "2008-04"; volume: "13"; issue: "4"; pages: "343-54"
COMMENT     Sequence Label: hRAD18-EGFP
FEATURES             Location/Qualifiers
     source          1..6311
                     /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      2251..2260
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2257..2259,2260..2262,2263..2976)
                     /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(2257..2259,2260..2262,2263..2973)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2302..2323)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2563..2582)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2910..2931
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3099..3220
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3136..3155)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3190..3209
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3227..3682)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3364..3385)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3576..3595
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3709..3813
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3815..4172
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3815..4006
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3838..3858)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4023..4158
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4207..5001
                     /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             4207..5001
                     /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(4261..4280)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4871..4890
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5189..5208)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5233..5280
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5609..6197
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6098..6117
                     /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 atctcgagat ccattgtggg cctgcgaggc ctagcgctag gcctcgcggg
      661 gaatttcgag tggtgttgga gcgccggagg ctagtgggtg gctgaccccc agcatcctcg
      721 ggagcgacca tggactccct ggccgagtct cggtggcctc cgggcctggc agtcatgaag
      781 acaatagatg atttgctgcg gtgtggaatt tgcttcgagt atttcaacat tgcaatgata
      841 atacctcagt gttcacataa ctactgctct ctctgtataa gaaaatttct gtcctataaa
      901 actcagtgtc caacttgctg tgtgactgtc acagagccgg atctgaaaaa taaccgcata
      961 ttagatgaac tggtaaaaag cttgaatttt gcacggaatc atctgctgca gtttgcttta
     1021 gagtcaccag ccaaatctcc tgcttcttcc tcttcaaaga atcttgctgt caaagtatat
     1081 actcctgtag cctccagaca gtctttaaag caggggagca ggttaatgga taatttcttg
     1141 atcagagaaa tgagtggttc tacatcagag ttgttgataa aagaaaataa aagcaaattc
     1201 agccctcaaa aagaggcgag ccctgctgca aagaccaaag agacacgttc tgtagaagag
     1261 atcgctccag atccctcaga ggctaagcgt cctgagccac cctcgacatc cactttgaaa
     1321 caagttacta aagtggattg tcctgtttgc ggggttaaca ttccagaaag tcacattaat
     1381 aagcatttag acagctgttt atcacgcgaa gagaagaagg aaagcctcag aagttctgtt
     1441 cacaaaagga agccgctgcc caaaactgta tataatttgc tctctgatcg tgatttaaag
     1501 aaaaagctaa aagagcatgg attatctatt caaggaaata aacaacagct cattaaaagg
     1561 caccaagaat ttgtacacat gtacaatgcc caatgcgatg ctttgcatcc taaatcagct
     1621 gctgaaatag ttcaagaaat cgaaaatata gagaagacta ggatgcgtct tgaagctagt
     1681 aaactcaatg aaagtgtaat ggtttttaca aaggaccaaa cagaaaagga aatagatgaa
     1741 atccacagta aatatcgtaa aaaacataag agtgaatttc agcttctggt ggatcaggct
     1801 agaaaaggat acaagaaaat tgctggaatg tcacaaaaaa cagtaacaat aacaaaagaa
     1861 gatgaatcta cagaaaagct atcttctgta tgcatgggac aggaagataa tatgacctca
     1921 gtaacaaacc acttttctca atcaaagctg gactccccag aggaattgga acctgacaga
     1981 gaagaggatt cttctagctg tattgatatt caagaagttc tttcttcatc agaatcagat
     2041 tcatgcaata gttccagttc agacatcata agagatcttt tagaagaaga ggaagcctgg
     2101 gaagcatcac ataaaaacga tcttcaagac acagaaataa gtccaagaca gaatcgccgc
     2161 acaagagccg ctgaaagtgc tgagattgaa ccaagaaaca agcgtaatag gaattctgca
     2221 gtcgacggta ccgcgggccc gggatccatc gccaccatgg tgagcaaggg cgaggagctg
     2281 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     2341 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     2401 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     2461 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     2521 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     2581 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     2641 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     2701 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     2761 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     2821 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     2881 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     2941 gggatcactc tcggcatgga cgagctgtac aagtaaagcg gccgcgactc tagatcataa
     3001 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     3061 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     3121 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3181 attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt
     3241 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     3301 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     3361 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     3421 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     3481 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     3541 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     3601 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     3661 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     3721 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     3781 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     3841 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3901 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3961 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4021 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4081 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     4141 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     4201 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4261 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4321 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4381 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4441 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4501 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4561 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4621 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4681 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     4741 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4801 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4861 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4921 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4981 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     5041 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     5101 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     5161 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     5221 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     5281 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     5341 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     5401 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     5461 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5521 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5581 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5641 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5701 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     5761 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5821 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5881 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5941 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6001 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6061 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6121 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6181 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6241 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     6301 ccgccatgca t
//
