LOCUS       sequence-473174-        6494 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 6494)
  TITLE     GMCL1 Controls 53BP1 Stability and Modulates Paclitaxel Sensitivity 
            in Cancer
REFERENCE   2  (bases 1 to 6494)
  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_GMCL1BBO-FLAG
FEATURES             Location/Qualifiers
     source          1..6494
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(27..43)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(27..43)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(40..62)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    51..67
                     /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(75..81,82..99,100..105))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    120..141
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(258..275)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(429..1017)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(509..528)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1188..1979,1980..2048))
                     /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"
     primer_bind     1811..1830
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2049..2153)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2228..2247)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2419..2798
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2799..3002
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2953..2973
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3047..3066
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3047..3065
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4038..4061
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4077..4100
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4116..4139
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             4176..4199
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     promoter        complement(4231..4249)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4232..4249)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4269..4286)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4275..4499
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4545..4973
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4632..4651)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4842..4863
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4982..5002)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4987..5316
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5167..5302
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5229..5248
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5383..6177
                     /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(5437..5456)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6047..6066
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6351..6472
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6388..6407)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6442..6461
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
       61 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
      121 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
      181 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
      241 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
      301 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
      361 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
      421 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
      481 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
      541 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
      601 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
      661 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
      721 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
      781 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
      841 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
      901 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
      961 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     1021 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     1081 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     1141 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     1201 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     1261 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     1321 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     1381 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     1441 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     1501 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     1561 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     1621 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     1681 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     1741 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     1801 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     1861 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     1921 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     1981 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     2041 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     2101 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     2161 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca
     2221 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     2281 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     2341 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
     2401 gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
     2461 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     2521 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     2581 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     2641 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     2701 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     2761 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     2821 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     2881 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     2941 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     3001 ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat acgactcact
     3061 atagggagac ccaagcttgg taccgagctc ggatccacta gtaacggccg ccagtgtgct
     3121 ggatgggatc gttgagcagc cgggtgctgc gccagccaag accagccctt gcccagcagg
     3181 cgcagggtgc cagggcgggg ggctcggccc ggaggccgga cactggagac gatgcggcgg
     3241 gccacggatt ctgttactgt gcgggcagcc acaagcgcaa gcggagcagc gggtccttct
     3301 gctactgtca ccctgactcg gagacggacg aggatgagga ggagggggac gagcagcagc
     3361 ggctcctcaa cacccctcga aggaaaaaat taaagagtac atctaaatat atttatcaaa
     3421 cattattttt gaatggtgaa aacagtgaca ttaagatttg tgctctagga gaagaatgga
     3481 gcttacacaa aatatattta tgtcaatctg gctacttttc tagtatgttc agtggttctt
     3541 ggaaagaatc cagcatgaat attattgaac tggagattcc tgaccagaac attgatgtag
     3601 aagcactgca ggttgcattt ggttcactgt atcgagatga tgtcttgata aagcccagtc
     3661 gagttgttgc cattttggca gcagcttgtt tgctgcagtt ggacggttta atacagcagt
     3721 gtggtgagac aatgaaggaa acagttaatg tgaaaactgt atgtggctat tacacatcag
     3781 cagggaccta tggattagat tctgtaaaga aaaagtgcct tgaatggctt ctaaacaatt
     3841 tgatgactca ccagaatgtt gaacttttta aagaactcag tataaatgtc atgaaacagc
     3901 tcattggttc atctaactta tttgtgatgc aagtggagat ggatatatac actgctctaa
     3961 aaaagtggat gttccttcaa cttgtgcctt cttggaatgg atctttaaaa cagcttttga
     4021 caggaggatc aggaatggac tacaaggacg acgatgacaa ggtaccagga tctatggatt
     4081 ataaagatga tgatgataaa gggtcggccg ccagctggag ccaccctcag ttcgagaagg
     4141 gaggaggaag cggcggaggc agcggaggag gaagctggag ccacccgcag ttcgagaaag
     4201 gagctggatc ctgagtctag agggccctat tctatagtgt cacctaaatg ctagagctcg
     4261 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     4321 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     4381 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     4441 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     4501 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     4561 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4621 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4681 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4741 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4801 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4861 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     4921 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     4981 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5041 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     5101 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     5161 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     5221 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     5281 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     5341 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     5401 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     5461 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     5521 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     5581 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5641 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5701 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5761 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5821 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5881 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     5941 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     6001 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     6061 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     6121 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     6181 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     6241 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     6301 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     6361 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     6421 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     6481 gtataccgtc gacc
//
