LOCUS       sequence-211197-        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  JOURNAL   Unpublished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            "Unpublished"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"
     enhancer        150..453
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        454..657
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     608..628
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      696..705
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(702..704,705..707,708..1418)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(702..704,705..707,708..1418)
                     /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(747..768)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1008..1027)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1355..1376
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5863..5984
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5900..5919)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5954..5973
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5991..6446)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6128..6149)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6340..6359
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6473..6577
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6579..6936
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6579..6770
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6602..6622)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6787..6922
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6971..7765
                     /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             6971..7765
                     /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(7025..7044)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7635..7654
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7953..7972)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7997..8044
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8373..8961
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8862..8881
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
       61 ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca
      121 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      181 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      241 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      301 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      361 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      421 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      481 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      541 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      601 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt
      661 ttagtgaacc gtcagatccg ctagcgctac cggtcgccac catggtgagc aagggcgagg
      721 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
      781 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
      841 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
      901 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
      961 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     1021 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     1081 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     1141 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     1201 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     1261 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     1321 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     1381 ccgccgggat cactctcggc atggacgagc tgtacaagtc cggactcaga tctcgagctc
     1441 ttatggctgc tgttcctcaa aataatctac aggagcaact agaacgtcac tcagccagaa
     1501 cacttaataa taaattaagt ctttcaaaac caaaattttc aggtttcact tttaaaaaga
     1561 aaacatcttc agataacaat gtatctgtaa ctaatgtgtc agtagcaaaa acacctgtat
     1621 taagaaataa agatgttaat gttaccgaag acttttcctt cagtgaacct ctacccaaca
     1681 ccacaaatca gcaaagggtc aaggacttct ttaaaaatgc tccagcagga caggaaacac
     1741 agagaggtgg atcaaaatca ttattgccag atttcttgca gactccgaag gaagttgtat
     1801 gcactaccca aaacacacca actgtaaaga aatcccggga tactgctctc aagaaattag
     1861 aatttagttc ttcaccagat tctttaagta ccatcaatga ttgggatgat atggatgact
     1921 ttgatacttc tgagacttca aaatcatttg ttacaccacc ccaaagtcac tttgtaagag
     1981 taagcactgc tcagaaatca aaaaagggta agagaaactt ttttaaagca cagctttata
     2041 caacaaacac agtaaagact gacttgcctc caccctcctc tgaaagcgag caaatagatt
     2101 tgactgagga acagaaggat gactcagaat ggttaagcag cgatgtgatt tgcatcgatg
     2161 atggccccat tgctgaagtg catataaatg aagatgctca ggaaagtgac tctctgaaaa
     2221 ctcatttgga agatgaaaga gataatagcg aaaagaagaa gaatttggaa gaagctgaat
     2281 tacattcaac tgagaaagtt ccatgtattg aatttgatga tgatgattat gatacggatt
     2341 ttgttccacc ttctccagaa gaaattattt ctgcttcttc ttcctcttca aaatgcctta
     2401 gtacgttaaa ggaccttgac acctctgaca gaaaagagga tgttcttagc acatcaaaag
     2461 atcttttgtc aaaacctgag aaaatgagta tgcaggagct gaatccagaa accagcacag
     2521 actgtgacgc tagacagata agtttacagc agcagcttat tcatgtgatg gagcacatct
     2581 gtaaattaat tgatactatt cctgatgata aactgaaact tttggattgt gggaacgaac
     2641 tgcttcagca gcggaacata agaaggaaac ttctaacgga agtagatttt aataaaagtg
     2701 atgccagtct tcttggctca ttgtggagat acaggcctga ttcacttgat ggccctatgg
     2761 agggtgattc ctgccctaca gggaattcta tgaaggagtt aaatttttca caccttccct
     2821 caaattctgt ttctcctggg gactgtttac tgactaccac cctaggaaag acaggattct
     2881 ctgccaccag gaagaatctt tttgaaaggc ctttattcaa tacccattta cagaagtcct
     2941 ttgtaagtag caactgggct gaaacaccaa gactaggaaa aaaaaatgaa agctcttatt
     3001 tcccaggaaa tgttctcaca agcactgctg tgaaagatca gaataaacat actgcttcaa
     3061 taaatgactt agaaagagaa acccaacctt cctatgatat tgataatttt gacatagatg
     3121 actttgatga tgatgatgac tgggaagaca taatgcataa tttagcagcc agcaaatctt
     3181 ccacagctgc ctatcaaccc atcaaggaag gtcggccaat taaatcagta tcagaaagac
     3241 tttcctcagc caagacagac tgtcttccag tgtcatctac tgctcaaaat ataaacttct
     3301 cagagtcaat tcagaattat actgacaagt cagcacaaaa tttagcatcc agaaatctga
     3361 aacatgagcg tttccaaagt cttagttttc ctcatacaaa ggaaatgatg aagatttttc
     3421 ataaaaaatt tggcctgcat aattttagaa ctaatcagct agaggcgatc aatgctgcac
     3481 tgcttggtga agactgtttt atcctgatgc cgactggagg tggtaagagt ttgtgttacc
     3541 agctccctgc ctgtgtttct cctggggtca ctgttgtcat ttctcccttg agatcactta
     3601 tcgtagatca agtccaaaag ctgacttcct tggatattcc agctacatat ctgacaggtg
     3661 ataagactga ctcagaagct acaaatattt acctccagtt atcaaaaaaa gacccaatca
     3721 taaaactcct atatgtcact ccagaaaaga tctgtgcaag taacagactc atttctactc
     3781 tggagaatct ctatgagagg aagctcttgg cacgttttgt tattgatgaa gcacattgtg
     3841 tcagtcagtg gggacatgat tttcgtcaag attacaaaag aatgaatatg cttcgccaga
     3901 agtttccttc tgttccggtg atggctctta cggccacagc taatcccagg gtacagaagg
     3961 acatcctgac tcagctgaag attctcagac ctcaggtgtt tagcatgagc tttaacagac
     4021 ataatctgaa atactatgta ttaccgaaaa agcctaaaaa ggtggcattt gattgcctag
     4081 aatggatcag aaagcaccac ccatatgatt cagggataat ttactgcctc tccaggcgag
     4141 aatgtgacac catggctgac acgttacaga gagatgggct cgctgctctt gcttaccatg
     4201 ctggcctcag tgattctgcc agagatgaag tgcagcagaa gtggattaat caggatggct
     4261 gtcaggttat ctgtgctaca attgcatttg gaatggggat tgacaaaccg gacgtgcgat
     4321 ttgtgattca tgcatctctc cctaaatctg tggagggtta ctaccaagaa tctggcagag
     4381 ctggaagaga tggggaaata tctcactgcc tgcttttcta tacctatcat gatgtgacca
     4441 gactgaaaag acttataatg atggaaaaag atggaaacca tcatacaaga gaaactcact
     4501 tcaataattt gtatagcatg gtacattact gtgaaaatat aacagaatgc aggagaatac
     4561 agcttttggc ctactttggt gaaaatggat ttaatcctga tttttgtaag aaacacccag
     4621 atgtttcttg tgataattgc tgtaaaacaa aggattataa aacaagagat gtgactgacg
     4681 atgtgaaaag tattgtaaga tttgttcaag aacatagttc atcacaagga atgagaaata
     4741 taaaacatgt aggtccttct ggaagattta ctatgaatat gctggtcgac attttcttgg
     4801 ggagtaagag tgcaaaaatc cagtcaggta tatttggaaa aggatctgct tattcacgac
     4861 acaatgccga aagacttttt aaaaagctga tacttgacaa gattttggat gaagacttat
     4921 atatcaatgc caatgaccag gcgatcgctt atgtgatgct cggaaataaa gcacaaactg
     4981 tactaaatgg caatttaaag gtagacttta tggaaacaga aaattccagc agtgtgaaaa
     5041 aacaaaaagc gttagtagca aaagtgtctc agagggaaga gatggttaaa aaatgtcttg
     5101 gagaacttac agaagtctgc aaatctctgg ggaaagtttt tggtgtccat tacttcaata
     5161 tttttaatac cgtcactctc aagaagcttg cagaatcttt atcttctgat cctgaggttt
     5221 tgcttcaaat tgatggtgtt actgaagaca aactggaaaa atatggtgcg gaagtgattt
     5281 cagtattaca gaaatactct gaatggacat cgccagctga agacagttcc ccagggataa
     5341 gcctgtccag cagcagaggc cccggaagaa gtgccgctga ggagcttgac gaggaaatac
     5401 ccgtatcttc ccactacttt gcaagtaaaa ccagaaatga aaggaagagg aaaaagatgc
     5461 cagcctccca aaggtctaag aggagaaaaa ctgcttccag tggttccaag gcaaaggggg
     5521 ggtctgccac atgtagaaag atatcttcca aaacgaaatc ctccagcatc attggatcca
     5581 gttcagcctc acatacttct caagcgacat caggagccaa tagcaaattg gggattatgg
     5641 ctccaccgaa gcctataaat agaccgtttc ttaagccttc atatgcattc tcataacaac
     5701 cgaatctcaa tgctcgaggg gctagagcgg ccgctcgagc atgcatctag ataactgatc
     5761 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     5821 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     5881 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     5941 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt aaattgtaag
     6001 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     6061 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     6121 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     6181 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     6241 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     6301 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     6361 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     6421 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
     6481 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     6541 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg
     6601 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     6661 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     6721 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     6781 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     6841 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     6901 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg
     6961 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     7021 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     7081 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     7141 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     7201 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     7261 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     7321 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     7381 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     7441 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
     7501 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     7561 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     7621 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     7681 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     7741 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     7801 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     7861 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     7921 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa
     7981 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt
     8041 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     8101 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     8161 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac
     8221 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     8281 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     8341 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     8401 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     8461 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     8521 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     8581 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     8641 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     8701 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     8761 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     8821 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     8881 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     8941 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttac
//
