LOCUS       sequence-423157-        9110 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP-tagged BLM helicase in mammalian cells (EGFP-C1 
            backbone).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9110)
  AUTHORS   Hu P, Beresten SF, van Brabant AJ, Ye TZ, Pandolfi PP, Johnson FB, 
            Guarente L, Ellis NA
  TITLE     Evidence for BLM and Topoisomerase IIIalpha interaction in genomic 
            stability.
  JOURNAL   Hum Mol Genet. 2001 Jun 1;10(12):1287-98.
  PUBMED    11406610
REFERENCE   2  (bases 1 to 9110)
  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: "Hum Mol 
            Genet."; date: "2001-06-1"; volume: "10(12)"; pages: "1287-98"
COMMENT     Sequence Label: GFP-BLM
FEATURES             Location/Qualifiers
     source          1..9110
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        35..338
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        339..542
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     493..513
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      581..590
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(587..589,590..592,593..1303)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(587..589,590..592,593..1303)
                     /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(632..653)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(893..912)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1240..1261
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1349..1360
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5872..5993
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5909..5928)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5963..5982
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(6000..6455)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6137..6158)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6349..6368
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6482..6586
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6588..6945
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6588..6779
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6611..6631)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6796..6931
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6980..7774
                     /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             6980..7774
                     /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(7034..7053)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7644..7663
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7962..7981)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    8006..8053
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8382..8970
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8871..8890
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
       61 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      121 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      181 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      241 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      301 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
      361 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      421 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
      481 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
      541 ctggtttagt gaaccgtcag atccgctagc gctaccggtc gccaccatgg tgagcaaggg
      601 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
      661 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
      721 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
      781 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
      841 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
      901 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
      961 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     1021 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     1081 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     1141 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     1201 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     1261 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtccggac tcagatctcg
     1321 agctcaagct tcgaattctg cagtcgagat cgagggtaga atggctgctg ttcctcaaaa
     1381 taatctacag gagcaactag aacgtcactc agccagaaca cttaataata aattaagtct
     1441 ttcaaaacca aaattttcag gtttcacttt taaaaagaaa acatcttcag ataacaatgt
     1501 atctgtaact aatgtgtcag tagcaaaaac acctgtatta agaaataaag atgttaatgt
     1561 taccgaagac ttttccttca gtgaacctct acccaacacc acaaatcagc aaagggtcaa
     1621 ggacttcttt aaaaatgctc cagcaggaca ggaaacacag agaggtggat caaaatcatt
     1681 attgccagat ttcttgcaga ctccgaagga agttgtatgc actacccaaa acacaccaac
     1741 tgtaaagaaa tcccgggata ctgctctcaa gaaattagaa tttagttctt caccagattc
     1801 tttaagtacc atcaatgatt gggatgatat ggatgacttt gatacttctg agacttcaaa
     1861 atcatttgtt acaccacccc aaagtcactt tgtaagagta agcactgctc agaaatcaaa
     1921 aaagggtaag agaaactttt ttaaagcaca gctttataca acaaacacag taaagactga
     1981 tttgcctcca ccctcctctg aaagcgagca aatagatttg actgaggaac agaaggatga
     2041 ctcagaatgg ttaagcagcg atgtgatttg catcgatgat ggccccattg ctgaagtgca
     2101 tataaatgaa gatgctcagg aaagtgactc tctgaaaact catttggaag atgaaagaga
     2161 taatagcgaa aagaagaaga atttggaaga agctgaatta cattcaactg agaaagttcc
     2221 atgtattgaa tttgatgatg atgattatga tacggatttt gttccacctt ctccagaaga
     2281 aattatttct gcttcttctt cctcttcaaa atgccttagt acgttaaagg accttgacac
     2341 atctgacaga aaagaggatg ttcttagcac atcaaaagat cttttgtcaa aacctgagaa
     2401 aatgagtatg caggagctga atccagaaac cagcacagac tgtgacgcta gacagataag
     2461 tttacagcag cagcttattc atgtgatgga gcacatctgt aaattaattg atactattcc
     2521 tgatgataaa ctgaaacttt tggattgtgg gaacgaactg cttcagcagc ggaacataag
     2581 aaggaaactt ctaacggaag tagattttaa taaaagtgat gccagtcttc ttggctcatt
     2641 gtggagatac aggcctgatt cacttgatgg ccctatggag ggtgattcct gccctacagg
     2701 gaattctatg aaggagttaa atttttcaca ccttccctca aattctgttt ctcctgggga
     2761 ctgtttactg actaccaccc taggaaagac aggattctct gccaccagga agaatctttt
     2821 tgaaaggcct ttattcaata cccatttaca gaagtccttt gtaagtagca actgggctga
     2881 aacaccaaga ctaggaaaaa aaaatgaaag ctcttatttc ccaggaaatg ttctcacaag
     2941 cactgctgtg aaagatcaga ataaacatac tgcttcaata aatgacttag aaagagaaac
     3001 ccaaccttcc tatgatattg ataattttga catagatgac tttgatgatg atgatgactg
     3061 ggaagacata atgcataatt tagcagccag caaatcttcc acagctgcct atcaacccat
     3121 caaggaaggt cggccaatta aatcagtatc agaaagactt tcctcagcca agacagactg
     3181 tcttccagtg tcatctactg ctcaaaatat aaacttctca gagtcaattc agaattatac
     3241 tgacaagtca gcacaaaatt tagcatccag aaatctgaaa catgagcgtt tccaaagtct
     3301 tagttttcct catacaaagg aaatgatgaa gatttttcat aaaaaatttg gcctgcataa
     3361 ttttagaact aatcagctag aggcgatcaa tgctgcactg cttggtgaag actgttttat
     3421 cctgatgccg actggaggtg gtaagagttt gtgttaccag ctccctgcct gtgtttctcc
     3481 tggggtcact gttgtcattt ctcccttgag atcacttatc gtagatcaag tccaaaagct
     3541 gacttccttg gatattccag ctacatatct gacaggtgat aagactgact cagaagctac
     3601 aaatatttac ctccagttat caaaaaaaga cccaatcata aaacttctat atgtcactcc
     3661 agaaaagatc tgtgcaagta acagactcat ttctactctg gagaatctct atgagaggaa
     3721 gctcttggca cgttttgtta ttgatgaagc acattgtgtc agtcagtggg gacatgattt
     3781 tcgtcaagat tacaaaagaa tgaatatgct tcgccagaag tttccttctg ttccggtgat
     3841 ggctcttacg gccacagcta atcccagggt acagaaggac atcctgactc agctgaagat
     3901 tctcagacct caggtgttta gcatgagctt taacagacat aatctgaaat actatgtatt
     3961 accgaaaaag cctaaaaagg tggcatttga ttgcctagaa tggatcagaa agcaccaccc
     4021 atatgattca gggataattt actgcctctc caggcgagaa tgtgacacca tggctgacac
     4081 gttacagaga gatgggctcg ctgctcttgc ttaccatgct ggcctcagtg attctgccag
     4141 agatgaagtg cagcagaagt ggattaatca ggatggctgt caggttatct gtgctacaat
     4201 tgcatttgga atggggattg acaaaccgga cgtgcgattt gtgattcatg catctctccc
     4261 taaatctgtg gagggttact accaagaatc tggcagagct ggaagagatg gggaaatatc
     4321 tcactgcctg cttttctata cctatcatga tgtgaccaga ctgaaaagac ttataatgat
     4381 ggaaaaagat ggaaaccatc atacaagaga aactcacttc aataatttgt atagcatggt
     4441 acattactgt gaaaatataa cggaatgcag gagaatacag cttttggcct actttggtga
     4501 aaatggattt aatcctgatt tttgtaagaa acacccagat gtttcttgtg ataattgctg
     4561 taaaacaaag gattataaaa caagagatgt gactgacgat gtgaaaagta ttgtaagatt
     4621 tgttcaagaa catagttcat cacaaggaat gagaaatata aaacatgtag gtccttctgg
     4681 aagatttact atgaatatgc tggtcgacat tttcttgggg agtaagagtg caaaaatcca
     4741 gtcaggtata tttggaaaag gatctgctta ttcacgacac aatgccgaaa gactttttaa
     4801 aaagctgata cttgacaaga ttttggatga agacttatat atcaatgcca atgaccaggc
     4861 gatcgcttat gtgatgctcg gaaataaagc ccaaactgta ctaaatggca atttaaaggt
     4921 agactttatg gaaacagaaa attccagcag tgtgaaaaaa caaaaagcgt tagtagcaaa
     4981 agtgtctcag agggaagaga tggttaaaaa atgtcttgga gaacttacag aagtctgcaa
     5041 atctctgggg aaagtttttg gtgtccatta cttcaatatt tttaataccg tcactctcaa
     5101 gaagcttgca gaatctttat cttctgatcc tgaggttttg cttcaaattg atggtgttac
     5161 tgaagacaaa ctggaaaaat atggtgcgga agtgatttca gtattacaga aatactctga
     5221 atggacatcg ccagctgaag acagttcccc agggataagc ctgtccagca gcagaggccc
     5281 cggaagaagt gccgctgagg agctcgacga ggaaatacct gtatcttccc actactttgc
     5341 aagtaaaacc agaaatgaaa ggaagaggaa aaagatgcca gcctcccaaa ggtctaagag
     5401 gagaaaaact gcttccagtg gttccaaggc aaaggggggg tctgccacat gtagaaagat
     5461 atcttccaaa acgaaatcct ccagcatcat tggatccagt tcagcctcac atacttctca
     5521 agcgacatca ggagccaata gcaaattggg gattatggct ccaccgaagc ctataaatag
     5581 accgtttctt aagccttcat atgcattctc ataacaaccg aatctcaatg tacatagacc
     5641 ctctttcttg tttgtcagca tctgaccatc tgtgactata aagctgttat tcttgttata
     5701 ccaaaaaaaa aaaaaaaaaa actcgacggt accgcgggcc cgggatccac cggatctaga
     5761 taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     5821 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     5881 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     5941 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta
     6001 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     6061 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     6121 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     6181 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     6241 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     6301 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     6361 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     6421 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     6481 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     6541 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
     6601 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     6661 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     6721 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     6781 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     6841 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     6901 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     6961 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     7021 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     7081 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     7141 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     7201 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     7261 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     7321 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     7381 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     7441 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     7501 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     7561 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     7621 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     7681 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     7741 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     7801 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     7861 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     7921 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     7981 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     8041 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     8101 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     8161 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     8221 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     8281 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     8341 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     8401 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     8461 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     8521 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     8581 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     8641 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     8701 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     8761 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     8821 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     8881 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     8941 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     9001 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     9061 gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg
//
