LOCUS       sequence-402918-        5395 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MCL-1 phosphodegron mutants.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5395)
  AUTHORS   Keyan KS, Alanany R, Kohil A, Khan OM
  TITLE     E3 Ubiquitin Ligase TRIP12 Controls Exit from Mitosis via Positive 
            Regulation of MCL-1 in Response to Taxol.
  JOURNAL   Cancers (Basel). 2023 Jan 13;15(2):505. doi: 
            10.3390/cancers15020505.
  PUBMED    36672454
REFERENCE   2  (bases 1 to 5395)
  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: "Cancers 
            (Basel)."; date: "2023-01-13"; volume: "15(2):505. doi"; pages: " 
            10.3390/cancers15020505"
COMMENT     Sequence Label: Flag- MCL1-3A (S159, S162, T163 mutated to A)
FEATURES             Location/Qualifiers
     source          1..5395
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        554..572
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             616..639
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1758..1774)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1759..1775)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1830..1849)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1831..1849)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2123..2244
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2160..2179)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2214..2233
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2251..2706)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2388..2409)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2600..2619
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2733..2837
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2839..3196
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2839..3030
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2862..2882)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3047..3182
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3231..4025
                     /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             3231..4025
                     /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(3285..3304)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3895..3914
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4213..4232)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4257..4304
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4633..5221
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5122..5141
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgatt
      541 acgccaagct cgaaattaac cctcactaaa gggaacaaaa gctggagctc caccgcggtg
      601 gcggccgcca ccatggatta caaggatgac gacgataagg cccgggcgga tccactagtc
      661 cagtgtggtg gaattgccct tatgtttggc ctcaaaagaa acgcggtaat cggactcaac
      721 ctctactgtg ggggggccgg cttgggggcc ggcagcggcg gcgccacccg cccgggaggg
      781 cgacttttgg ctacggagaa ggaggcctcg gcccggcgag agataggggg aggggaggcc
      841 ggcgcggtga ttggcggaag cgccggcgca agccccccgt ccaccctcac gccagactcc
      901 cggagggtcg cgcggccgcc gcccattggc gccgaggtcc ccgacgtcac cgcgaccccc
      961 gcgaggctgc ttttcttcgc gcccacccgc cgcgcggcgc cgcttgagga gatggaagcc
     1021 ccggccgctg acgccatcat gtcgcccgaa gaggagctgg acgggtacga gccggagcct
     1081 ctcgggaagc ggccggctgt cctgccgctg ctggagttgg tcggggaatc tggtaataac
     1141 accagtacgg acggggcact acccgcggcg ccgccgccag cagaggagga ggacgagttg
     1201 taccggcagt cgctggagat tatctctcgg taccttcggg agcaggccac cggcgccaag
     1261 gacacaaagc caatgggcag gtctggggcc accagcagga aggcgctgga gaccttacga
     1321 cgggttgggg atggcgtgca gcgcaaccac gagacggcct tccaaggcat gcttcggaaa
     1381 ctggacatca aaaacgaaga cgatgtgaaa tcgttgtctc gagtgatgat ccatgttttc
     1441 agcgacggcg taacaaactg gggcaggatt gtgactctca tttcttttgg tgcctttgtg
     1501 gctaaacact tgaagaccat aaaccaagaa agctgcatcg aaccattagc agaaagtatc
     1561 acagacgttc tcgtaaggac aaaacgggac tggctagtta aacaaagagg ctgggatggg
     1621 tttgtggagt tcttccatgt agaggaccta gaaggtggca tcaggaatgt gctgctggct
     1681 tttgcaggtg ttgctggagt aggagctggt ttggcatatc taataagata gggcaattct
     1741 gcagatatca agcttatcga taccgtcgac ctcgaggggg ggcccggtac cttaattaat
     1801 taaggtacca ggtaagtgta cccaattcgc cctatagtga gtcgtattac aattcactcg
     1861 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gagatccaat ttttaagtgt
     1921 ataatgtgtt aaactactga ttctaattgt ttgtgtattt tagattcaca gtcccaaggc
     1981 tcatttcagg cccctcagtc ctcacagtct gttcatgatc ataatcagcc ataccacatt
     2041 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     2101 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     2161 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     2221 gtccaaactc atcaatgtat cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat
     2281 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     2341 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     2401 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     2461 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     2521 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     2581 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     2641 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     2701 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     2761 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2821 attgaaaaag gaagaatcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     2881 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     2941 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3001 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3061 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3121 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     3181 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3241 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3301 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3361 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     3421 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     3481 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     3541 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     3601 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     3661 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gaacatcagg
     3721 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     3781 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     3841 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     3901 ctacccgtga tattgctgaa gaacttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     3961 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4021 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4081 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4141 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     4201 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     4261 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     4321 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     4381 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     4441 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     4501 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     4561 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     4621 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     4681 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     4741 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     4801 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4861 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4921 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4981 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     5041 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     5101 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     5161 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     5221 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     5281 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt
     5341 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccg
//
