LOCUS       sequence-473590-        7059 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 7059)
  TITLE     GMCL1 Controls 53BP1 Stability and Modulates Paclitaxel Sensitivity 
            in Cancer
REFERENCE   2  (bases 1 to 7059)
  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_GMCL2-FLAG
FEATURES             Location/Qualifiers
     source          1..7059
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     361..380
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(599..703)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(778..797)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        969..1348
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1349..1552
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1503..1523
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1597..1616
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1597..1615
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1678..1687
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3274..3297
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3310..3333
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(3346..3364)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3347..3364)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3384..3401)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3390..3614
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3660..4088
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3747..3766)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3957..3978
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4097..4117)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4102..4431
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4282..4417
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4344..4363
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4498..5292
                     /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(4552..4571)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5162..5181
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5466..5587
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5503..5522)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5557..5576
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5636..5652)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5636..5652)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5649..5671)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5660..5676
                     /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(5684..5690,5691..5708,5709..5714))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5729..5750
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5867..5884)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6038..6626)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6118..6137)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6797..7059,1..529,530..598))
                     /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 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
       61 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
      121 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
      181 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
      241 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
      301 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
      361 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
      421 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
      481 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
      541 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
      601 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
      661 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
      721 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac
      781 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
      841 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
      901 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata
      961 tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
     1021 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     1081 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     1141 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     1201 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     1261 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     1321 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
     1381 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     1441 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
     1501 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct
     1561 aactagagaa cccactgctt actggcttat cgaaattaat acgactcact atagggagac
     1621 ccaagcttgg taccgagctc ggatccacta gtaacggccg ccagtgtgct ggaattcgcc
     1681 accatgggat cgtcgagcag ccgggtgctg ggccagccga ggcgagccct tgcccagcag
     1741 gaacagggtg ccagggccag gggctcggcc cggaggccgg acactggaga cgatgcggcg
     1801 agctacggct tctgttactg cccgggcagt cacaagcgca agcggagcag cggggcctgc
     1861 cgctactgtg acccggactc gcacagggag gagcatgagg aggaggggga caagcagcag
     1921 ccgctcctca acacccctgc aaggaaaaaa ttaaggagta catccaaata tatttatcaa
     1981 acattatttt tgaatggtga aaacagtgac attaagattt gtgctctagg agaagaatgg
     2041 cgattacaca aaatatattt atgtcaatct ggctactttt ctagtatgtt cagtggttct
     2101 tggaaagaat ccagcatgaa tattattgaa ctggagattc ctgaccagaa cattgatgta
     2161 gacgcactgc aggttgcgtt tggttcactg tatcgagatg atgtcttgat aaaacccagt
     2221 cgagttgttg ccattttggc agcagcttgt atgctgcagc tggatggttt aatacagcag
     2281 tgtggtgaga caatgaagga aacaattaat gtgaaaactg tatgcggtta ttacacatca
     2341 gtagagatct atggattaga ttctgtaaag aaaaagtgcc ttgaatggct tctaaacaat
     2401 ttgatgactc accagaatgt taaacttttt aaagaactcg gtataaatgt catgaaacag
     2461 ctcattggtt cctctaactt atttgtgatg caagtggaga tggatgtata caccactcta
     2521 aaaaagtgga tgttccttca acttgtgcct tcttggaatg gatctttaaa acagcttttg
     2581 acagaaacag atgtctggtt ttctaaacag agaaaagatt ttgaaggtat ggcctttctt
     2641 gaaactgaac caggaaaacc atttgtgtca gtattcagac atttaaggtt acaatatatt
     2701 atcagtgacc tagcttctgc aagaattatt gaacaagatg gtatagtacc ttcagaatgg
     2761 ctgtcttctg tgtataaaca gcagtggttt gctatgctgc gggcagaaca agaccgtgag
     2821 gtagggcctc aagaaatcaa taaagaagac ctagagggaa atagcatgag gtgtggtaga
     2881 aagcttgcca aagatggtga atactactgg tgttggacgg gttttaactt cggctttgac
     2941 ctacttgtaa tttacaccaa tggatacatc attttcaaac gcaatacact gaatcagcca
     3001 cgcagcgggt ctgtcagttt acggcctcga aggagcatag catttagatt acgcttggct
     3061 tcttttgata gtagtggaaa actagtatgt agtagaacaa ctggctatca aatacttata
     3121 cttaaaaagg atcaggaaca agtggtgatg aacttggaca gcaggtttct gaccttccct
     3181 ttatatatct gctgtaactt cttgtatata tcaccagaaa aaggaattga aaataatcgt
     3241 cacccagaaa atccagaaaa cggaggatca ggagactaca aggacgacga tgacaaggga
     3301 ggatcaggag attataaaga tgatgatgat aaatgagggc cctattctat agtgtcacct
     3361 aaatgctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     3421 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     3481 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     3541 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     3601 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     3661 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3721 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3781 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3841 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3901 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3961 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     4021 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4081 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4141 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4201 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4261 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4321 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     4381 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     4441 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     4501 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4561 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4621 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     4681 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4741 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4801 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4861 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4921 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4981 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     5041 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     5101 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     5161 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     5221 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     5281 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     5341 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     5401 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     5461 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5521 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5581 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     5641 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     5701 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5761 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5821 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5881 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5941 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6001 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6061 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6121 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6181 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6241 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6301 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6361 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6421 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6481 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6541 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6601 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6661 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6721 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6781 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6841 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6901 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6961 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7021 tatccgcctc catccagtct attaattgtt gccgggaag
//
