LOCUS       sequence-487966-        8487 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_053, derived from human fusion protein DOC2B_USP43.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8487)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8487)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 2, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: CL20_mEGFP_FOidr_053
FEATURES             Location/Qualifiers
     source          1..8487
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     181..200
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    199..254
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     300..319
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(join(2020..2730,2731..2733,2734..2736))
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2062..2083)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2411..2430
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2670..2691
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2733..2742
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2818..3021)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2847..2867)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3022..3325
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3582..3626)
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    3811..4044
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    4404..4521
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5013..5138
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5185..5442
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5444..5823
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5854..5958
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5959..6027,6028..6819)
                     /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(6177..6196)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6990..7578
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7479..7498
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7732..7749
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7817..7951
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7841..7860)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7895..7914
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7957..8152)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7971..8106
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    8211..8232
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8247..8252,8253..8270,8271..8277)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8285..8301
                     /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     8290..8312
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8309..8325
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8309..8325
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aaaaaaaatc taacctcaag tcaaggcttt tctatggaat aaggaatgga cagcaggggg
       61 ctgtttcata tactgatgac ctctttatag ccaacctttg ttcatggcag ccagcatatg
      121 ggcatatgtt gccaaactct aaaccaaata ctcattctga tgttttaaat gatttgccct
      181 cccatatgtc cttccgagtg agagacacaa aaaattccaa cacactattg caatgaaaat
      241 aaatttcctt tattagccag aagtcagatg ctcaaggggc ttcatgatgt ccccataatt
      301 tttggcagag ggaaaaagat ctgctagctc gagcagctga agctttattg aggcttaagc
      361 agtgggttcc ctagttagcc agagagctcc caggctcaga tctggtctaa ccagagagac
      421 ccagtacagg caaaaagcag atcttgtctt ctttgggagt gaattagccc ttccagtccc
      481 cccttttctt ttaaaaagtg gctaagatct acagctgcct tgtaagtcat tggtcttaaa
      541 ggtaccgtaa tcttatctag aaccggttca gaacgaactc tctggaagag tctttttgcg
      601 tcccaatgat aacgtgtggt acttaacgcg ctgaaatgtc ccgtagctag cctgctcacc
      661 gggaacggta cgctgggaca ttcctaaggc agagccacgt ggtgcacggg aggctttgcg
      721 cggaagacgt aatgagctcg gggcactcca aacatcacgc tcaaggcgac taccaagtcc
      781 acgagccaac ccctctggaa tgcgcagcgc cggcggtgac ccaggtgctt gcccatccat
      841 tgcgggagaa agacctccag acaccaagga taccggtggg acttgaggtg acacttctgc
      901 acgctcgttg cggcgatttt cctttttgcg aaagacacta cgcagaagcg taagggtccc
      961 ctgaagggga cgatctaact cactcgtacc acgacgcgag tccttagaat tgccggaaaa
     1021 tcccatgtat ggtgaaggtg gactgctttt agaacccatc tgaaatgact ccttccagtt
     1081 cattgctttt tgcccctcgg ggcgcgcagg tttttcatgt tcaacactag gcataacagt
     1141 caagggcagg tcaaacttac gcggcagttt aatgtcctga cctgcatttt ctttcagctt
     1201 acgcgcagtg ttcgggccat cactattacc tggtgcaagg caatgatttg gttgcgcgga
     1261 tggacatggg actccggcag gtccacgctc gatagcgcga cccccgtcct cagaatttgc
     1321 gggtaatgcg acattggagc gaggggaggc ggatttttcg tcttctcccg cggtgcctgt
     1381 aagcagggac acgattgatt gggatgtcga acgagggcgg cgacgggatt ccaagtactc
     1441 caccagctcg acgctagcac ctggcggctt ttctttcgac ccgaaactcc agcgtaatgg
     1501 catggtcttg aagggcccct gctttgcgcg tgacgtcgta ggtgccttca tcgaaatcga
     1561 gcggccttta accccacgaa ctaaacggcg aggctccagg ccgcgacagt catcggattc
     1621 atatccgtcg gcatcgggtt cgtcttcggg gggcttagca gggggacgag caggagaggg
     1681 accagggcta ggccctggtg aagacccgta tgctccgaac agttggtcaa catcctcgtc
     1741 gtcctcacgc gctccatctg aagggctacg gcgaccagct ccggctacgg caggacgagc
     1801 tggcgcatct gggggtgcag ccgcacgggg acccgcgtct ggtggcaagc cacggggaaa
     1861 acgggggaaa taatctgaaa tttgtttaat ggggcgaata ggaccagggc atacgtcaat
     1921 tgccatgtgc tcttggatac taatcgttgc tttttctcca cggcgacgta acgtcatccg
     1981 ggaagcggcc gcacttcccc cggagcctcc gcttccgccc ttgtacagct cgtccatgcc
     2041 gagagtgatc ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt
     2101 ggggtctttg ctcagcttgg actgggtgct caggtagtgg ttgtcgggca gcagcacggg
     2161 gccgtcgccg atgggggtgt tctgctggta gtggtcggcg agctgcacgc tgccgtcctc
     2221 gatgttgtgg cggatcttga agttcacctt gatgccgttc ttctgcttgt cggccatgat
     2281 atagacgttg tggctgttgt agttgtactc cagcttgtgc cccaggatgt tgccgtcctc
     2341 cttgaagtcg atgcccttca gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac
     2401 ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct cctggacgta
     2461 gccttcgggc atggcggact tgaagaagtc gtgctgcttc atgtggtcgg ggtagcggct
     2521 gaagcactgc acgccgtagg tcagggtggt cacgagggtg ggccagggca cgggcagctt
     2581 gccggtggtg cagatgaact tcagggtcag cttgccgtag gtggcatcgc cctcgccctc
     2641 gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca ggatgggcac
     2701 caccccggtg aacagctcct cgcccttgct caccatggtg gcgaccggtg gatcccgggc
     2761 ccgcggtacc gtcgactgca gaattcggtt cactaaacga gcccggttca ctaaacgagc
     2821 tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca atggggcgga
     2881 gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac tcccattgac
     2941 gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac gcccattgat
     3001 gtactgccaa aaccgcatca ccatggtaat agcgatgact aatacgtaga tgtactgcca
     3061 agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc
     3121 attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc
     3181 agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat
     3241 gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg
     3301 gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa ctaatgaccc
     3361 cgtaattgat tactattaat aactagcacg cgttcgaagc ttgagctcgg tcccctcggg
     3421 gttgggaggt gggtctgaaa cgataatggt gaatatccct gcctaactct attcactata
     3481 gaaagtacag caaaaactat tcttaaacct accaagcctc ctactatcat tatgaataat
     3541 tttatatacc acagccaatt tgttatgtta aaccaattcc acaaacttgc ccatttatct
     3601 aattccaata attcttgttc attcttttct tgctggtttt gcgattcttc aattaaggag
     3661 tgtattaagc ttgtgtaatt gttaatttct ctgtcccact ccatccaggt cgtgtgattc
     3721 caaatctgtt ccagagattt attactccaa ctagcattcc aaggcacagc agtggtgcaa
     3781 atgagttttc cagagcaacc ccaaatcccc aggagctgtt gatcctttag gtatctttcc
     3841 acagccagga ttcttgcctg gagctgcttg atgccccaga ctgtgagttg caacagatgc
     3901 tgttgcgcct caatagccct cagcaaattg ttctgctgct gcactatacc agacaataat
     3961 tgtctggcct gtaccgtcag cgtcattgac gctgcgccca tagtgcttcc tgctgctccc
     4021 aagaacccaa ggaacaaagc tcctattccc actgctcttt tttctctctg caccactctt
     4081 ctctttgcct tggtgggtgc tactcctaat ggttcaattt ttactacttt atatttatat
     4141 aattcacttc tccaattgtc cctcatatct cctccttatg gtgttttact aaacttttcc
     4201 atgttctaat cctcatcctg tctacttgcc acacaatcat cacctgccat ctgttttcca
     4261 taatccctaa tgatctttgc ttttcttctt ggcactactt ttatgtcact attatcttgt
     4321 attactactg ccccttcacc tttccagagg agctttgctg gtcctttcca aagtggattt
     4381 ctgctgtccc tgtaataaac ccgaaaattt tgaatttttg taatttgttt ttgtaattct
     4441 ttagtttgta tgtctgttgc tattatgtct actattcttt cccctgcact gtacccccca
     4501 atcccccctt ttcttttaaa attgtggatg aatactgcca tttgtactgc tgtcttaaga
     4561 tgttcagcct gatctcttac ctgtcctata attttcttta attctttatt catagattct
     4621 actactcctt gactttgggg attgtaggga atttccaggt ctgaagatcc tgctgcttgc
     4681 tgtgcttttt tcttactttt gttttgctct tcctctatct tgtctaaagc ttccttggtg
     4741 tcttttatct ctatcctttg atgcacacaa tagagggttg ctactgtatt atataatgat
     4801 ctaagttctt ctgatcctgt ctgaagggat ggttgtagct gtcccagtat ttgtctacag
     4861 ccttctgatg tttctaacag gccagtatta actgcgaatc gttctagctc cctgcttgcc
     4921 catactatat gttttaattt atattttttc tttccccctg gccttaaccg aattttttcc
     4981 catcgcgatc taattctccc ccgcttaata ctgacgctct cgcacccatc tctctccttc
     5041 tagcctccgc tagtcaaaat ttttggcgta ctcaccagtc gccgcccctc gcctcttgcc
     5101 gtgcgcgctt cagcaagccg agtcctgcgt cgagagagct cctctggttt ccctttcgct
     5161 ttcaagtccc tgttcgggcg ccactgctag agattttcca cactgactaa aagggtctga
     5221 gggatctcta gttaccagag tcacacaaca gacgggcaca cactacttga agcactcaag
     5281 gcaagcttta ttgaggctta agcagtgggt tccctagtta gccagagagc tcccaggctc
     5341 agatctggtc taaccagaga gacccagtac aggcaaaaag cagctgctta tatgcaggat
     5401 ctgagggctc gccactcccc agtcccgccc aggccacgcc tcccatggta atagcgatga
     5461 ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat
     5521 gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca
     5581 cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga
     5641 aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt
     5701 cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat
     5761 atgggctatg aactaatgac cccgtaattg attactatta ataactagtc aataatcaat
     5821 gtcgacccag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     5881 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5941 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     6001 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     6061 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     6121 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     6181 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     6241 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     6301 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     6361 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     6421 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     6481 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     6541 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     6601 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     6661 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     6721 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     6781 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     6841 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6901 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6961 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     7021 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     7081 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     7141 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     7201 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     7261 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     7321 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     7381 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     7441 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     7501 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     7561 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     7621 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     7681 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     7741 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     7801 attcattaat gcagcggatc cagacatgat aagatacatt gatgagtttg gacaaaccac
     7861 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     7921 tgtaaccatt ataagctgca ataaacaagt tagctttttg caaaagccta ggcctccaaa
     7981 aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg cctctgcata
     8041 aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg
     8101 cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat gcggatccgc
     8161 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     8221 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     8281 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     8341 cttgggctgc aggtcgaggg atctccataa gagaagaggg acagctatga ctgggagtag
     8401 tcaggagagg aggaaaaatc tggctagtaa aacatgtaag gaaaatttta gggatgttaa
     8461 agaaaaaaat aacacaaaac aaaatat
//
