LOCUS       sequence-420517-        9269 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of human TopBP1 N-terminally fused to mVenus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9269)
  TITLE     Pospiech Lab Plasmids 2023
REFERENCE   2  (bases 1 to 9269)
  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"
COMMENT     Sequence Label: pmVen-TopBP1
FEATURES             Location/Qualifiers
     source          1..9269
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        88..391
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        392..595
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     546..566
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      634..643
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(640..642,643..645,646..1356)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(685..706)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(946..965)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1293..1314
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(5003..5014)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     polyA_signal    6084..6205
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6121..6140)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6175..6194
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(6212..6667)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6349..6370)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6561..6580
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6694..6798
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6800..7157
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6800..6991
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6823..6843)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      7008..7143
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             7192..7986
                     /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             7192..7986
                     /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(7246..7265)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7856..7875
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(8174..8193)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    8218..8265
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8594..9182
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9083..9102
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt
       61 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
      121 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
      181 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
      241 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
      301 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
      361 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
      421 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
      481 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
      541 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt
      601 agtgaaccgt cagatccgct agcgctaccg gtcgccacca tggtgagcaa gggcgaggag
      661 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
      721 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagctg
      781 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgggctac
      841 ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
      901 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
      961 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     1021 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     1081 agccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag
     1141 atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca gcagaacacc
     1201 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccaag
     1261 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     1321 gccgggatca ctctcggcat ggacgagctg tacaagtccg gactcagatc tcgagctcac
     1381 atgtccagaa atgacaaaga accgtttttt gtgaagtttt taaagtcttc agacaattcc
     1441 aaatgttttt ttaaagctct cgagtccata aaagaattcc aatcagaaga atatcttcag
     1501 attattacag aagaagaggc attgaagata aaggagaatg atagatcact ttatatctgt
     1561 gaccctttta gtggcgttgt ctttgatcac ctcaaaaagc ttggctgcag aattgttggt
     1621 cctcaagtag tcatattttg tatgcaccac cagcgatgtg tcccaagagc cgaacatcca
     1681 gtttataata tggttatgtc tgatgtaacc atatcttgta caagtctgga aaaagaaaaa
     1741 agggaagaag ttcataaata tgtacaaatg atgggcggac gagtatacag agaccttaat
     1801 gtatcagtaa ctcaccttat tgcaggagaa gttggtagca aaaaatattt agttgctgca
     1861 aacctgaaga aacctatttt gcttccctct tggataaaaa cactttggga gaagtcacaa
     1921 gagaaaaaaa taactagata tactgatata aacatggaag atttcaagtg tcctattttt
     1981 cttggttgca taatctgtgt gactggctta tgtggcttag acaggaaaga agttcagcaa
     2041 ctcacagtta agcatggagg tcaatacatg ggacaattga aaatgaatga atgtacacac
     2101 ctcattgtgc aagaaccaaa aggtcagaag tatgagtgtg ccaagagatg gaatgtacac
     2161 tgtgtgacca cacagtggtt ttttgacagt attgagaaag gtttttgtca ggatgaatcc
     2221 atatacaaga cagaacctag accagaagca aagactatgc ccaattcttc aactcctacc
     2281 agccagatca acacaattga tagtcgtact ctttcagatg tcagcaatat ttccaacata
     2341 aatgcaagtt gcgtaagtga atcaatatgt aattcactta acagcaaact ggagcctaca
     2401 cttgaaaatc tagaaaatct ggatgtcagt gcatttcaag cacctgaaga tttattagat
     2461 ggttgtcgga tatatctttg cggttttagt ggcagaaagc tagataaact gagaagactt
     2521 attaacagtg gaggtggagt tcgttttaac cagctaaatg aagatgtaac tcatgttatt
     2581 gtgggagatt atgatgatga attgaagcag ttttggaata aatcagccca caggcctcat
     2641 gtagtgggag caaagtggtt gctagagtgt ttcagtaaag gttatatgct ttctgaagaa
     2701 ccatatatcc atgctaatta ccagccagtg gaaattccag tttcacatca gcctgaaagt
     2761 aaagcagctc ttttaaaaaa gaagaacagc agcttctcta agaaagactt tgctcctagt
     2821 gaaaagcatg agcaagctga tgaagatctg ctctctcaat atgaaaatgg tagctccaca
     2881 gtagttgagg ctaagacgtc tgaagccagg ccctttaatg attctactca tgctgagccc
     2941 ttgaatgatt ctactcacat ttctttgcaa gaagaaaacc agtcttctgt cagtcattgt
     3001 gtccctgatg tttctacaat tactgaagaa ggcttattta gccaaaagag tttccttgtt
     3061 ttgggtttta gtaatgaaaa tgaatctaac atcgcaaaca tcataaaaga aaatgctggg
     3121 aaaatcatgt cccttctgag cagaactgtt gcggattatg ctgtggttcc tctgctgggg
     3181 tgtgaagtgg aagccactgt gggagaagtt gttacaaata catggctggt tacttgcata
     3241 gactatcaga ctttgtttga tccaaagtcg aatcctctct tcacaccagt tccagtaatg
     3301 acaggaatga ctcctttaga ggattgtgtt atttcattta gccagtgtgc tggagcagaa
     3361 aaagagtctt taactttcct agcaaacctc cttggagcaa gtgttcaaga atactttgtt
     3421 cgcaaatcca atgcaaagaa aggcatgttt gccagtactc atcttatact gaaagaacgt
     3481 ggtggctcta aatatgaagc tgcaaagaag tggaatttac ctgccgttac tatagcttgg
     3541 ctgttggaga ctgctagaac gggaaagaga gcagacgaaa gccattttct gattgaaaat
     3601 tcaactaaag aagaacgaag tttggaaaca gaaataacaa atggaatcaa tctaaattca
     3661 gatactgcag agcatcctgg cacacgcctg caaactcaca gaaaaaccgt cgttacacct
     3721 ttagatatga accgctttca gagtaaagct ttccgtgctg tggtctcaca acatgccaga
     3781 caggtcgcag cctccccagc agtaggacaa ccacttcaga aggagccctc gttacacctg
     3841 gatacaccat caaaattcct gtccaaggac aaactcttca agccttcctt tgatgtgaag
     3901 gatgcacttg cagccttgga aactccagga cgtcccagcc aacagaaaag gaaaccgagt
     3961 acgccactct cagaagttat tgtcaaaaac ttgcaacttg ctttggcaaa tagctctcga
     4021 aatgctgtcg ctctttctgc cagccctcaa ctgaaagagg cccagtcaga gaaggaagaa
     4081 gccccaaagc cacttcacaa agtagtggta tgtgttagta aaaaactcag taagaagcag
     4141 agtgaactaa atgggatcgc agcctctcta ggagcagatt acaggtggag ttttgatgaa
     4201 acagtgactc atttcatcta tcaagggcgg ccaaatgaca ctaatcggga gtataaatct
     4261 gtaaaagaaa gaggagtaca cattgtttcc gagcactggc ttttagattg tgcccaagag
     4321 tgtaaacatc ttcctgaatc tctttatcca catacttata atcccaaaat gagcttggat
     4381 atcagcgcag tgcaagatgg ccggctctgt aatagtcgac tactctcagc tgtgtcttca
     4441 acaaaggatg atgagccaga tcctttgatt ttagaagaaa atgatgtaga caatatggcc
     4501 accaataata aagagtcagc accatcaaat ggaagtggaa agaatgactc taaaggagtt
     4561 ctgacacaga ccttagagat gagagagaac tttcagaagc agttacagga gataatgtct
     4621 gcaacatcaa tagtgaaacc ccaagggcag aggacttccc tttcaagaag tggttgtaac
     4681 agcgcatctt caacccctga cagcactcgc tctgctcgca gtggacgaag tagagtccta
     4741 gaggcactga ggcagtctcg tcagacagta cctgatgtca acacagagcc ttcccaaaat
     4801 gaacagatca tttgggatga ccctacagca agggaggaga gagcaaggct tgccagcaat
     4861 ttgcagtggc ctagttgtcc cacacaatac tctgagcttc aggttgacat tcaaaacttg
     4921 gaggattctc cttttcaaaa gcctttacat gattcagaaa ttgctaaaca ggctgtctgt
     4981 gatcctggaa acatacgtgt gactgaagct cccaaacacc caatctctga agaactggaa
     5041 actcccataa aagacagcca cctgatccct acgcctcaag cccccagtat tgcctttcca
     5101 ctcgccaacc cccctgtggc tccgcaccct agagaaaaga ttataacgat agaggagact
     5161 catgaagaat taaaaaaaca gtacatattt cagttatcat ctctgaatcc tcaagaacgt
     5221 attgactatt gtcatctgat tgagaaacta ggtggattgg tgatagaaaa gcagtgcttt
     5281 gatcccacct gtacacacat tgttgtggga catccacttc gaaacgagaa gtatttagcc
     5341 tcagtggcag ctgggaagtg ggtgcttcat cgctcctacc ttgaagcctg caggactgct
     5401 ggacacttcg tgcaggaaga agactatgaa tggggaagta gttccatact tgatgttctg
     5461 actggaatca atgtacagca acgaagacta gcacttgcag caatgagatg gagaaaaaaa
     5521 atccagcaaa gacaagaatc tggcattgtt gagggagcat ttagtgggtg gaaggttatt
     5581 ttacatgtgg atcagtctcg agaagcaggc ttcaaacgcc ttcttcagtc aggaggagca
     5641 aaggtgctac ctggtcattc tgtaccttta tttaaagagg ccacacatct tttttctgac
     5701 ttgaataaac tgaaaccaga tgactcagga gttaatatag cagaagctgc tgcccagaac
     5761 gtgtactgct tgagaacaga atacattgct gattatctca tgcaggaatc acctcctcat
     5821 gtagaaaatt actgtctacc agaagctatt tcatttattc agaataataa ggaacttggg
     5881 actggattat cacaaaagag gaaagctcct acagaaaaaa ataaaatcaa acgacctaga
     5941 gtacactaat cgcaggatcc accggatcta gataactgat cataatcagc cataccacat
     6001 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     6061 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     6121 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     6181 tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa
     6241 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     6301 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     6361 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     6421 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     6481 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     6541 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     6601 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     6661 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     6721 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     6781 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     6841 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     6901 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     6961 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     7021 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     7081 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     7141 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     7201 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     7261 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     7321 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     7381 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     7441 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     7501 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     7561 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     7621 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     7681 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     7741 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     7801 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     7861 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     7921 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     7981 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     8041 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     8101 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     8161 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     8221 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     8281 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     8341 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     8401 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     8461 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     8521 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     8581 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     8641 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     8701 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     8761 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     8821 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     8881 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     8941 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     9001 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     9061 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     9121 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     9181 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     9241 tgttctttcc tgcgttatcc cctgattct
//
