LOCUS       sequence-420515-        9269 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of human TopBP1 N-terminally fused to 
            mCerulean.
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: pmCer-TopBP1
FEATURES             Location/Qualifiers
     source          1..9269
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        64..367
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        368..571
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     522..542
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      610..619
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(616..618,619..621,622..1332)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(661..682)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(922..941)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1269..1290
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(4979..4990)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     polyA_signal    6060..6181
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6097..6116)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6151..6170
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(6188..6643)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6325..6346)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6537..6556
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6670..6774
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6776..7133
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6776..6967
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6799..6819)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6984..7119
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             7168..7962
                     /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             7168..7962
                     /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(7222..7241)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7832..7851
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(8150..8169)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    8194..8241
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8570..9158
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9059..9078
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cattagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
       61 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      121 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      181 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      241 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      301 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      361 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
      421 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      481 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
      541 tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg
      601 ctaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
      661 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
      721 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
      781 gtgccctggc ccaccctcgt gaccaccctg acctggggcg tgcagtgctt cgcccgctac
      841 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
      901 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
      961 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     1021 aacatcctgg ggcacaagct ggagtacaac gccatcagcg acaacgtcta tatcaccgcc
     1081 gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc
     1141 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     1201 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     1261 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     1321 gagctgtaca agtccggact cagatctcga gctcacatgt ccagaaatga caaagaaccg
     1381 ttttttgtga agtttttaaa gtcttcagac aattccaaat gtttttttaa agctctcgag
     1441 tccataaaag aattccaatc agaagaatat cttcagatta ttacagaaga agaggcattg
     1501 aagataaagg agaatgatag atcactttat atctgtgacc cttttagtgg cgttgtcttt
     1561 gatcacctca aaaagcttgg ctgcagaatt gttggtcctc aagtagtcat attttgtatg
     1621 caccaccagc gatgtgtccc aagagccgaa catccagttt ataatatggt tatgtctgat
     1681 gtaaccatat cttgtacaag tctggaaaaa gaaaaaaggg aagaagttca taaatatgta
     1741 caaatgatgg gcggacgagt atacagagac cttaatgtat cagtaactca ccttattgca
     1801 ggagaagttg gtagcaaaaa atatttagtt gctgcaaacc tgaagaaacc tattttgctt
     1861 ccctcttgga taaaaacact ttgggagaag tcacaagaga aaaaaataac tagatatact
     1921 gatataaaca tggaagattt caagtgtcct atttttcttg gttgcataat ctgtgtgact
     1981 ggcttatgtg gcttagacag gaaagaagtt cagcaactca cagttaagca tggaggtcaa
     2041 tacatgggac aattgaaaat gaatgaatgt acacacctca ttgtgcaaga accaaaaggt
     2101 cagaagtatg agtgtgccaa gagatggaat gtacactgtg tgaccacaca gtggtttttt
     2161 gacagtattg agaaaggttt ttgtcaggat gaatccatat acaagacaga acctagacca
     2221 gaagcaaaga ctatgcccaa ttcttcaact cctaccagcc agatcaacac aattgatagt
     2281 cgtactcttt cagatgtcag caatatttcc aacataaatg caagttgcgt aagtgaatca
     2341 atatgtaatt cacttaacag caaactggag cctacacttg aaaatctaga aaatctggat
     2401 gtcagtgcat ttcaagcacc tgaagattta ttagatggtt gtcggatata tctttgcggt
     2461 tttagtggca gaaagctaga taaactgaga agacttatta acagtggagg tggagttcgt
     2521 tttaaccagc taaatgaaga tgtaactcat gttattgtgg gagattatga tgatgaattg
     2581 aagcagtttt ggaataaatc agcccacagg cctcatgtag tgggagcaaa gtggttgcta
     2641 gagtgtttca gtaaaggtta tatgctttct gaagaaccat atatccatgc taattaccag
     2701 ccagtggaaa ttccagtttc acatcagcct gaaagtaaag cagctctttt aaaaaagaag
     2761 aacagcagct tctctaagaa agactttgct cctagtgaaa agcatgagca agctgatgaa
     2821 gatctgctct ctcaatatga aaatggtagc tccacagtag ttgaggctaa gacgtctgaa
     2881 gccaggccct ttaatgattc tactcatgct gagcccttga atgattctac tcacatttct
     2941 ttgcaagaag aaaaccagtc ttctgtcagt cattgtgtcc ctgatgtttc tacaattact
     3001 gaagaaggct tatttagcca aaagagtttc cttgttttgg gttttagtaa tgaaaatgaa
     3061 tctaacatcg caaacatcat aaaagaaaat gctgggaaaa tcatgtccct tctgagcaga
     3121 actgttgcgg attatgctgt ggttcctctg ctggggtgtg aagtggaagc cactgtggga
     3181 gaagttgtta caaatacatg gctggttact tgcatagact atcagacttt gtttgatcca
     3241 aagtcgaatc ctctcttcac accagttcca gtaatgacag gaatgactcc tttagaggat
     3301 tgtgttattt catttagcca gtgtgctgga gcagaaaaag agtctttaac tttcctagca
     3361 aacctccttg gagcaagtgt tcaagaatac tttgttcgca aatccaatgc aaagaaaggc
     3421 atgtttgcca gtactcatct tatactgaaa gaacgtggtg gctctaaata tgaagctgca
     3481 aagaagtgga atttacctgc cgttactata gcttggctgt tggagactgc tagaacggga
     3541 aagagagcag acgaaagcca ttttctgatt gaaaattcaa ctaaagaaga acgaagtttg
     3601 gaaacagaaa taacaaatgg aatcaatcta aattcagata ctgcagagca tcctggcaca
     3661 cgcctgcaaa ctcacagaaa aaccgtcgtt acacctttag atatgaaccg ctttcagagt
     3721 aaagctttcc gtgctgtggt ctcacaacat gccagacagg tcgcagcctc cccagcagta
     3781 ggacaaccac ttcagaagga gccctcgtta cacctggata caccatcaaa attcctgtcc
     3841 aaggacaaac tcttcaagcc ttcctttgat gtgaaggatg cacttgcagc cttggaaact
     3901 ccaggacgtc ccagccaaca gaaaaggaaa ccgagtacgc cactctcaga agttattgtc
     3961 aaaaacttgc aacttgcttt ggcaaatagc tctcgaaatg ctgtcgctct ttctgccagc
     4021 cctcaactga aagaggccca gtcagagaag gaagaagccc caaagccact tcacaaagta
     4081 gtggtatgtg ttagtaaaaa actcagtaag aagcagagtg aactaaatgg gatcgcagcc
     4141 tctctaggag cagattacag gtggagtttt gatgaaacag tgactcattt catctatcaa
     4201 gggcggccaa atgacactaa tcgggagtat aaatctgtaa aagaaagagg agtacacatt
     4261 gtttccgagc actggctttt agattgtgcc caagagtgta aacatcttcc tgaatctctt
     4321 tatccacata cttataatcc caaaatgagc ttggatatca gcgcagtgca agatggccgg
     4381 ctctgtaata gtcgactact ctcagctgtg tcttcaacaa aggatgatga gccagatcct
     4441 ttgattttag aagaaaatga tgtagacaat atggccacca ataataaaga gtcagcacca
     4501 tcaaatggaa gtggaaagaa tgactctaaa ggagttctga cacagacctt agagatgaga
     4561 gagaactttc agaagcagtt acaggagata atgtctgcaa catcaatagt gaaaccccaa
     4621 gggcagagga cttccctttc aagaagtggt tgtaacagcg catcttcaac ccctgacagc
     4681 actcgctctg ctcgcagtgg acgaagtaga gtcctagagg cactgaggca gtctcgtcag
     4741 acagtacctg atgtcaacac agagccttcc caaaatgaac agatcatttg ggatgaccct
     4801 acagcaaggg aggagagagc aaggcttgcc agcaatttgc agtggcctag ttgtcccaca
     4861 caatactctg agcttcaggt tgacattcaa aacttggagg attctccttt tcaaaagcct
     4921 ttacatgatt cagaaattgc taaacaggct gtctgtgatc ctggaaacat acgtgtgact
     4981 gaagctccca aacacccaat ctctgaagaa ctggaaactc ccataaaaga cagccacctg
     5041 atccctacgc ctcaagcccc cagtattgcc tttccactcg ccaacccccc tgtggctccg
     5101 caccctagag aaaagattat aacgatagag gagactcatg aagaattaaa aaaacagtac
     5161 atatttcagt tatcatctct gaatcctcaa gaacgtattg actattgtca tctgattgag
     5221 aaactaggtg gattggtgat agaaaagcag tgctttgatc ccacctgtac acacattgtt
     5281 gtgggacatc cacttcgaaa cgagaagtat ttagcctcag tggcagctgg gaagtgggtg
     5341 cttcatcgct cctaccttga agcctgcagg actgctggac acttcgtgca ggaagaagac
     5401 tatgaatggg gaagtagttc catacttgat gttctgactg gaatcaatgt acagcaacga
     5461 agactagcac ttgcagcaat gagatggaga aaaaaaatcc agcaaagaca agaatctggc
     5521 attgttgagg gagcatttag tgggtggaag gttattttac atgtggatca gtctcgagaa
     5581 gcaggcttca aacgccttct tcagtcagga ggagcaaagg tgctacctgg tcattctgta
     5641 cctttattta aagaggccac acatcttttt tctgacttga ataaactgaa accagatgac
     5701 tcaggagtta atatagcaga agctgctgcc cagaacgtgt actgcttgag aacagaatac
     5761 attgctgatt atctcatgca ggaatcacct cctcatgtag aaaattactg tctaccagaa
     5821 gctatttcat ttattcagaa taataaggaa cttgggactg gattatcaca aaagaggaaa
     5881 gctcctacag aaaaaaataa aatcaaacga cctagagtac actaatcgca ggatccaccg
     5941 gatctagata actgatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     6001 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     6061 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     6121 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     6181 aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     6241 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     6301 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     6361 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     6421 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     6481 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     6541 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     6601 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg
     6661 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     6721 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag
     6781 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     6841 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     6901 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     6961 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     7021 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     7081 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat
     7141 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     7201 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     7261 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     7321 gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc
     7381 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     7441 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     7501 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     7561 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     7621 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     7681 gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     7741 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     7801 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     7861 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     7921 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     7981 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     8041 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     8101 gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga
     8161 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     8221 acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg
     8281 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc
     8341 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg
     8401 ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat ttttaattta
     8461 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     8521 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     8581 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     8641 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     8701 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     8761 tatcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     8821 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     8881 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     8941 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     9001 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     9061 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     9121 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     9181 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     9241 attctgtgga taaccgtatt accgccatg
//
