LOCUS       sequence-207648-        8986 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of a EGFP-tagged full length Bloom's syndrome 
            helicase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8986)
  TITLE     plasmids for Bloom's syndrome helicase and p53BP1
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8986)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pEGFP-BLM
FEATURES             Location/Qualifiers
     source          1..8986
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    3080..3201
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3117..3136)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3171..3190
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3208..3663)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3345..3366)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3557..3576
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3690..3794
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3796..4153
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3796..3987
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3819..3839)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4004..4139
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4188..4982
                     /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             4188..4982
                     /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(4242..4261)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4852..4871
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5170..5189)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5214..5261
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5590..6178
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6079..6098
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6353..6656
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6657..6860
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6811..6831
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      6899..6908
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6905..6907,6908..6910,6911..7621)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(6905..6907,6908..6910,6911..7621)
                     /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(6950..6971)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7211..7230)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7558..7579
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 aattctatga aggagttaaa tttttcacac cttccctcaa attctgtttc tcctggggac
       61 tgtttactga ctaccaccct aggaaagaca ggattctctg ccaccaggaa gaatcttttt
      121 gaaaggcctt tattcaatac ccatttacag aagtcctttg taagtagcaa ctgggctgaa
      181 acaccaagac taggaaaaaa aaatgaaagc tcttatttcc caggaaatgt tctcacaagc
      241 actgctgtga aagatcagaa taaacatact gcttcaataa atgacttaga aagagaaacc
      301 caaccttcct atgatattga taattttgac atagatgact ttgatgatga tgatgactgg
      361 gaagacataa tgcataattt agcagccagc aaatcttcca cagctgccta tcaacccatc
      421 aaggaaggtc ggccaattaa atcagtatca gaaagacttt cctcagccaa gacagactgt
      481 cttccagtgt catctactgc tcaaaatata aacttctcag agtcaattca gaattatact
      541 gacaagtcag cacaaaattt agcatccaga aatctgaaac atgagcgttt ccaaagtctt
      601 agttttcctc atacaaagga aatgatgaag atttttcata aaaaatttgg cctgcataat
      661 tttagaacta atcagctaga ggcgatcaat gctgcactgc ttggtgaaga ctgttttatc
      721 ctgatgccga ctggaggtgg taagagtttg tgttaccagc tccctgcctg tgtttctcct
      781 ggggtcactg ttgtcatttc tcccttgaga tcacttatcg tagatcaagt ccaaaagctg
      841 acttccttgg atattccagc tacatatctg acaggtgata agactgactc agaagctaca
      901 aatatttacc tccagttatc aaaaaaagac ccaatcataa aacttctata tgtcactcca
      961 gaaaagatct gtgcaagtaa cagactcatt tctactctgg agaatctcta tgagaggaag
     1021 ctcttggcac gttttgttat tgatgaagca cattgtgtca gtcagtgggg acatgatttt
     1081 cgtcaagatt acaaaagaat gaatatgctt cgccagaagt ttccttctgt tccggtgatg
     1141 gctcttacgg ccacagctaa tcccagggta cagaaggaca tcctgactca gctgaagatt
     1201 ctcagacctc aggtgtttag catgagcttt aacagacata atctgaaata ctatgtatta
     1261 ccgaaaaagc ctaaaaaggt ggcatttgat tgcctagaat ggatcagaaa gcaccaccca
     1321 tatgattcag ggataattta ctgcctctcc aggcgagaat gtgacaccat ggctgacacg
     1381 ttacagagag atgggctcgc tgctcttgct taccatgctg gcctcagtga ttctgccaga
     1441 gatgaagtgc agcagaagtg gattaatcag gatggctgtc aggttatctg tgctacaatt
     1501 gcatttggaa tggggattga caaaccggac gtgcgatttg tgattcatgc atctctccct
     1561 aaatctgtgg agggttacta ccaagaatct ggcagagctg gaagagatgg ggaaatatct
     1621 cactgcctgc ttttctatac ctatcatgat gtgaccagac tgaaaagact tataatgatg
     1681 gaaaaagatg gaaaccatca tacaagagaa actcacttca ataatttgta tagcatggta
     1741 cattactgtg aaaatataac ggaatgcagg agaatacagc ttttggccta ctttggtgaa
     1801 aatggattta atcctgattt ttgtaagaaa cacccagatg tttcttgtga taattgctgt
     1861 aaaacaaagg attataaaac aagagatgtg actgacgatg tgaaaagtat tgtaagattt
     1921 gttcaagaac atagttcatc acaaggaatg agaaatataa aacatgtagg tccttctgga
     1981 agatttacta tgaatatgct ggtcgacatt ttcttgggga gtaagagtgc aaaaatccag
     2041 tcaggtatat ttggaaaagg atctgcttat tcacgacaca atgccgaaag actttttaaa
     2101 aagctgatac ttgacaagat tttggatgaa gacttatata tcaatgccaa tgaccaggcg
     2161 atcgcttatg tgatgctcgg aaataaagcc caaactgtac taaatggcaa tttaaaggta
     2221 gactttatgg aaacagaaaa ttccagcagt gtgaaaaaac aaaaagcgtt agtagcaaaa
     2281 gtgtctcaga gggaagagat ggttaaaaaa tgtcttggag aacttacaga agtctgcaaa
     2341 tctctgggga aagtttttgg tgtccattac ttcaatattt ttaataccgt cactctcaag
     2401 aagcttgcag aatctttatc ttctgatcct gaggttttgc ttcaaattga tggtgttact
     2461 gaagacaaac tggaaaaata tggtgcggaa gtgatttcag tattacagaa atactctgaa
     2521 tggacatcgc cagctgaaga cagttcccca gggataagcc tgtccagcag cagaggcccc
     2581 ggaagaagtg ccgctgagga gcttgacgag gaaatacccg tatcttccca ctactttgca
     2641 agtaaaacca gaaatgaaag gaagaggaaa aagatgccag cctcccaaag gtctaagagg
     2701 agaaaaactg cttccagtgg ttccaaggca aagggggggt ctgccacatg tagaaagata
     2761 tcttccaaaa cgaaatcctc cagcatcatt ggatccagtt cagcctcaca tacttctcaa
     2821 gcgacatcag gagccaatag caaattgggg attatggctc caccgaagcc tataaataga
     2881 ccgtttctta agccttcata tgcattctca taacaaccga atctcaatgc tcgaggggct
     2941 agagcggccg ctcgagcatg catctagata actgatcata atcagccata ccacatttgt
     3001 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     3061 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     3121 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     3181 caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg
     3241 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     3301 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     3361 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     3421 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     3481 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     3541 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     3601 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     3661 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3721 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3781 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     3841 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3901 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3961 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     4021 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     4081 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     4141 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     4201 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     4261 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4321 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     4381 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     4441 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     4501 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     4561 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     4621 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     4681 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     4741 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     4801 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     4861 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     4921 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     4981 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     5041 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     5101 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
     5161 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     5221 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     5281 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     5341 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     5401 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
     5461 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     5521 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5581 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5641 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5701 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     5761 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5821 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5881 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5941 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     6001 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     6061 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     6121 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     6181 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     6241 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt
     6301 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     6361 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     6421 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     6481 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     6541 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     6601 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
     6661 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     6721 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     6781 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
     6841 aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc taccggtcgc
     6901 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     6961 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     7021 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     7081 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     7141 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     7201 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     7261 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     7321 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     7381 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     7441 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     7501 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     7561 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     7621 gtccggactc agatctcgag ctcttatggc tgctgttcct caaaataatc tacaggagca
     7681 actagaacgt cactcagcca gaacacttaa taataaatta agtctttcaa aaccaaaatt
     7741 ttcaggtttc acttttaaaa agaaaacatc ttcagataac aatgtatctg taactaatgt
     7801 gtcagtagca aaaacacctg tattaagaaa taaagatgtt aatgttaccg aagacttttc
     7861 cttcagtgaa cctctaccca acaccacaaa tcagcaaagg gtcaaggact tctttaaaaa
     7921 tgctccagca ggacaggaaa cacagagagg tggatcaaaa tcattattgc cagatttctt
     7981 gcagactccg aaggaagttg tatgcactac ccaaaacaca ccaactgtaa agaaatcccg
     8041 ggatactgct ctcaagaaat tagaatttag ttcttcacca gattctttaa gtaccatcaa
     8101 tgattgggat gatatggatg actttgatac ttctgagact tcaaaatcat ttgttacacc
     8161 accccaaagt cactttgtaa gagtaagcac tgctcagaaa tcaaaaaagg gtaagagaaa
     8221 cttttttaaa gcacagcttt atacaacaaa cacagtaaag actgatttgc ctccaccctc
     8281 ctctgaaagc gagcaaatag atttgactga ggaacagaag gatgactcag aatggttaag
     8341 cagcgatgtg atttgcatcg atgatggccc cattgctgaa gtgcatataa atgaagatgc
     8401 tcaggaaagt gactctctga aaactcattt ggaagatgaa agagataata gcgaaaagaa
     8461 gaagaatttg gaagaagctg aattacattc aactgagaaa gttccatgta ttgaatttga
     8521 tgatgatgat tatgatacgg attttgttcc accttctcca gaagaaatta tttctgcttc
     8581 ttcttcctct tcaaaatgcc ttagtacgtt aaaggacctt gacacatctg acagaaaaga
     8641 ggatgttctt agcacatcaa aagatctttt gtcaaaacct gagaaaatga gtatgcagga
     8701 gctgaatcca gaaaccagca cagactgtga cgctagacag ataagtttac agcagcagct
     8761 tattcatgtg atggagcaca tctgtaaatt aattgatact attcctgatg ataaactgaa
     8821 acttttggat tgtgggaacg aactgcttca gcagcggaac ataagaagga aacttctaac
     8881 ggaagtagat tttaataaaa gtgatgccag tcttcttggc tcattgtgga gatacaggcc
     8941 tgattcactt gatggcccta tggagggtga ttcctgccct acaggg
//
