LOCUS       sequence-473588-        7139 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient overexpression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7139)
  TITLE     GMCL1 Controls 53BP1 Stability and Modulates Paclitaxel Sensitivity 
            in Cancer
REFERENCE   2  (bases 1 to 7139)
  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: pcDNA3_GMCL1E142K-FLAG
FEATURES             Location/Qualifiers
     source          1..7139
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     193..212
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(431..535)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(610..629)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        801..1180
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1181..1384
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1335..1355
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1429..1448
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1429..1447
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3065..3088
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3104..3127
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3143..3166
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             3203..3226
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     promoter        complement(3258..3276)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3259..3276)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3296..3313)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3302..3526
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3572..4000
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3659..3678)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3869..3890
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4009..4029)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4014..4343
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4194..4329
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4256..4275
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4410..5204
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4464..4483)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5074..5093
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5378..5499
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5415..5434)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5469..5488
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5548..5564)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5548..5564)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5561..5583)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5572..5588
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5596..5602,5603..5620,5621..5626))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5641..5662
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5779..5796)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5950..6538)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6030..6049)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6709..7139,1..361,362..430))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
       61 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
      121 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
      181 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
      241 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
      301 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
      361 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
      421 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
      481 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
      541 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg
      601 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg
      661 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
      721 gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt
      781 acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac
      841 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
      901 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      961 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     1021 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     1081 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     1141 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     1201 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     1261 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     1321 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     1381 agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca
     1441 ctatagggag acccaagctt ggtaccgagc tcggatccac tagtaacggc cgccagtgtg
     1501 ctggatggga tcgttgagca gccgggtgct gcgccagcca agaccagccc ttgcccagca
     1561 ggcgcagggt gccagggcgg ggggctcggc ccggaggccg gacactggag acgatgcggc
     1621 gggccacgga ttctgttact gtgcgggcag ccacaagcgc aagcggagca gcgggtcctt
     1681 ctgctactgt caccctgact cggagacgga cgaggatgag gaggaggggg acgagcagca
     1741 gcggctcctc aacacccctc gaaggaaaaa attaaagagt acatctaaat atatttatca
     1801 aacattattt ttgaatggtg aaaacagtga cattaagatt tgtgctctag gagaagaatg
     1861 gagcttacac aaaatatatt tatgtcaatc tggctacttt tctagtatgt tcagtggttc
     1921 ttggaaaaaa tccagcatga atattattga actggagatt cctgaccaga acattgatgt
     1981 agaagcactg caggttgcat ttggttcact gtatcgagat gatgtcttga taaagcccag
     2041 tcgagttgtt gccattttgg cagcagcttg tttgctgcag ttggacggtt taatacagca
     2101 gtgtggtgag acaatgaagg aaacagttaa tgtgaaaact gtatgtggct attacacatc
     2161 agcagggacc tatggattag attctgtaaa gaaaaagtgc cttgaatggc ttctaaacaa
     2221 tttgatgact caccagaatg ttgaactttt taaagaactc agtataaatg tcatgaaaca
     2281 gctcattggt tcatctaact tatttgtgat gcaagtggag atggatatat acactgctct
     2341 aaaaaagtgg atgttccttc aacttgtgcc ttcttggaat ggatctttaa aacagctttt
     2401 gacagaaaca gatgtctggt tttctaaaca gaggaaagat tttgaaggta tggcctttct
     2461 tgaaactgaa caaggaaaac catttgtgtc agtattcaga catttaaggt tacaatatat
     2521 tatcagtgat ctggcttctg caagaattat tgaacaagat gctgtagtac cttcagaatg
     2581 gctctcttct gtgtataaac agcagtggtt tgctatgctg cgggcagaac aggacagtga
     2641 ggtggggcct caagaaatca ataaagaaga actagaggga aacagcatga ggtgtggtag
     2701 aaagcttgcc aaagatggtg aatactgctg gcgttggaca ggttttaact tcggcttcga
     2761 cctacttgta acttacacca atcgatacat cattttcaaa cgcaatacac tgaatcagcc
     2821 atgtagcgga tctgtcagtt tacagcctcg aaggagcata gcatttagat tacgtttggc
     2881 ttcttttgat agtagtggaa aactaatatg tagtagaaca actggctatc aaatacttac
     2941 acttgaaaag gatcaggaac aagtggtgat gaacttggac agcaggcttc tgatcttccc
     3001 tttatatatc tgctgtaact tcttgtatat atcaccagaa aaaaagaatg gaggatcagg
     3061 aatggactac aaggacgacg atgacaaggt accaggatct atggattata aagatgatga
     3121 tgataaaggg tcggccgcca gctggagcca ccctcagttc gagaagggag gaggaagcgg
     3181 cggaggcagc ggaggaggaa gctggagcca cccgcagttc gagaaaggag ctggatcctg
     3241 agtctagagg gccctattct atagtgtcac ctaaatgcta gagctcgctg atcagcctcg
     3301 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     3361 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     3421 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     3481 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     3541 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     3601 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3661 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3721 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3781 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3841 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3901 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     3961 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     4021 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4081 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4141 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4201 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4261 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     4321 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     4381 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     4441 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4501 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     4561 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     4621 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4681 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4741 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     4801 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     4861 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     4921 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     4981 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     5041 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     5101 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     5161 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     5221 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     5281 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5341 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     5401 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5461 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     5521 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5581 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5641 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5701 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5761 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5821 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5881 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5941 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6001 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6061 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6121 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6181 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6241 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6301 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6361 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6421 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6481 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6541 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6601 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6661 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6721 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6781 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6841 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6901 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6961 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7021 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7081 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcct
//
