LOCUS       sequence-473444-        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_GMCL1-FLAG
FEATURES             Location/Qualifiers
     source          1..7139
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     586..605
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(726..1055)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      740..875
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(794..813)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     1040..1060
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(1069..1497)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1179..1200)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1391..1410
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    1543..1767
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1756..1773
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        1793..1811
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1793..1810
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(1843..1866)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             complement(1903..1926)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             complement(1942..1965)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(1981..2004)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3621..3640)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3622..3640)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3685..3888)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3714..3734)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3889..4268
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4440..4459
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4534..4638
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4639..4707,4708..5499)
                     /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     complement(4857..4876)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5670..6258
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6159..6178
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6412..6429
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6546..6567
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6582..6587,6588..6605,6606..6612)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6620..6636
                     /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)."
     primer_bind     6625..6647
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6644..6660
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6644..6660
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    6709..6830
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6720..6739)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6774..6793
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(7004..7139,1..659))
                     /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(7115..7134)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca
       61 ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg
      121 tcgggcatgc gcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct
      181 tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg
      241 cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc
      301 attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc
      361 tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc
      421 acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc
      481 agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct
      541 gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg
      601 aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg
      661 cgaaacgatc ctcatcctgt ctcttgatca gatccgaaaa tggatataca agctcccggg
      721 agctttttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca
      781 gaggcagagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga
      841 gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg cgggactatg
      901 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
      961 tttccacacc tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg
     1021 gagcctgggg actttccaca ccctaactga cacacattcc acagaattaa ttcgcgttaa
     1081 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     1141 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     1201 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     1261 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     1321 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     1381 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     1441 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtggg
     1501 gataccccct agagccccag ctggttcttt ccgcctcaga agccatagag cccaccgcat
     1561 ccccagcatg cctgctattg tcttcccaat cctccccctt gctgtcctgc cccaccccac
     1621 cccccagaat agaatgacac ctactcagac aatgcgatgc aatttcctca ttttattagg
     1681 aaaggacagt gggagtggca ccttccaggg tcaaggaagg cacgggggag gggcaaacaa
     1741 cagatggctg gcaactagaa ggcacagtcg aggctgatca gcgagctcta gcatttaggt
     1801 gacactatag aatagggccc tctagactca ggatccagct cctttctcga actgcgggtg
     1861 gctccagctt cctcctccgc tgcctccgcc gcttcctcct cccttctcga actgagggtg
     1921 gctccagctg gcggccgacc ctttatcatc atcatcttta taatccatag atcctggtac
     1981 cttgtcatcg tcgtccttgt agtccattcc tgatcctcca ttcttttttt ctggtgatat
     2041 atacaagaag ttacagcaga tatataaagg gaagatcaga agcctgctgt ccaagttcat
     2101 caccacttgt tcctgatcct tttcaagtgt aagtatttga tagccagttg ttctactaca
     2161 tattagtttt ccactactat caaaagaagc caaacgtaat ctaaatgcta tgctccttcg
     2221 aggctgtaaa ctgacagatc cgctacatgg ctgattcagt gtattgcgtt tgaaaatgat
     2281 gtatcgattg gtgtaagtta caagtaggtc gaagccgaag ttaaaacctg tccaacgcca
     2341 gcagtattca ccatctttgg caagctttct accacacctc atgctgtttc cctctagttc
     2401 ttctttattg atttcttgag gccccacctc actgtcctgt tctgcccgca gcatagcaaa
     2461 ccactgctgt ttatacacag aagagagcca ttctgaaggt actacagcat cttgttcaat
     2521 aattcttgca gaagccagat cactgataat atattgtaac cttaaatgtc tgaatactga
     2581 cacaaatggt tttccttgtt cagtttcaag aaaggccata ccttcaaaat ctttcctctg
     2641 tttagaaaac cagacatctg tttctgtcaa aagctgtttt aaagatccat tccaagaagg
     2701 cacaagttga aggaacatcc acttttttag agcagtgtat atatccatct ccacttgcat
     2761 cacaaataag ttagatgaac caatgagctg tttcatgaca tttatactga gttctttaaa
     2821 aagttcaaca ttctggtgag tcatcaaatt gtttagaagc cattcaaggc actttttctt
     2881 tacagaatct aatccatagg tccctgctga tgtgtaatag ccacatacag ttttcacatt
     2941 aactgtttcc ttcattgtct caccacactg ctgtattaaa ccgtccaact gcagcaaaca
     3001 agctgctgcc aaaatggcaa caactcgact gggctttatc aagacatcat ctcgatacag
     3061 tgaaccaaat gcaacctgca gtgcttctac atcaatgttc tggtcaggaa tctccagttc
     3121 aataatattc atgctggatt ctttccaaga accactgaac atactagaaa agtagccaga
     3181 ttgacataaa tatattttgt gtaagctcca ttcttctcct agagcacaaa tcttaatgtc
     3241 actgttttca ccattcaaaa ataatgtttg ataaatatat ttagatgtac tctttaattt
     3301 tttccttcga ggggtgttga ggagccgctg ctgctcgtcc ccctcctcct catcctcgtc
     3361 cgtctccgag tcagggtgac agtagcagaa ggacccgctg ctccgcttgc gcttgtggct
     3421 gcccgcacag taacagaatc cgtggcccgc cgcatcgtct ccagtgtccg gcctccgggc
     3481 cgagcccccc gccctggcac cctgcgcctg ctgggcaagg gctggtcttg gctggcgcag
     3541 cacccggctg ctcaacgatc ccatccagca cactggcggc cgttactagt ggatccgagc
     3601 tcggtaccaa gcttgggtct ccctatagtg agtcgtatta atttcgataa gccagtaagc
     3661 agtgggttct ctagttagcc agagagctct gcttatatag acctcccacc gtacacgcct
     3721 accgcccatt tgcgtcaatg gggcggagtt gttacgacat tttggaaagt cccgttgatt
     3781 ttggtgccaa aacaaactcc cattgacgtc aatggggtgg agacttggaa atccccgtga
     3841 gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac cgcatcacca tggtaatagc
     3901 gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc
     3961 ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg
     4021 atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca
     4081 atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg
     4141 ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaactc
     4201 catatatggg ctatgaacta atgaccccgt aattgattac tattaataac tagtcaataa
     4261 tcaatgtcaa cgcgtatatc tggcccgtac atcgcgaagc agcgcaaaac gcctaaccct
     4321 aagcagattc ttcatgcaat tgtcggtcaa gccttgcctt gttgtagctt aaattttgct
     4381 cgcgcactac tcagcgacct ccaacacaca agcagggagc agatactggc ttaactatgc
     4441 ggcatcagag cagattgtac tgagagtgca ccatagggga tcgggagatc tcccgatccg
     4501 tcgacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4561 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4621 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4681 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     4741 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     4801 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     4861 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     4921 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     4981 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5041 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5101 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5161 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     5221 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     5281 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     5341 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     5401 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     5461 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     5521 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5581 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5641 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5701 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5761 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5821 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5881 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5941 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6001 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6061 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6121 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6181 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6241 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6301 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     6361 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     6421 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     6481 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     6541 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     6601 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     6661 catgattacg ccaagctcta gctagaggtc gacggtatac agacatgata agatacattg
     6721 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     6781 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt ggggtgggcg
     6841 aagaactcca gcatgagatc cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg
     6901 attccgaagc ccaacctttc atagaaggcg gcggtggaat cgaaatctcg tgatggcagg
     6961 ttgggcgtcg cttggtcggt catttcgaac cccagagtcc cgctcagaag aactcgtcaa
     7021 gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga
     7081 agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatg
//
