LOCUS       sequence-488133-        7308 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_110, derived from human fusion protein MSH6_FBXO11.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7308)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7308)
  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_110
FEATURES             Location/Qualifiers
     source          1..7308
                     /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(841..1551,1552..1554,1555..1557))
                     /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(883..904)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1232..1251
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1491..1512
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1554..1563
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1639..1842)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1668..1688)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1843..2146
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2403..2447)
                     /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    2632..2865
                     /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    3225..3342
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    3834..3959
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4006..4263
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4265..4644
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4675..4779
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4780..4848,4849..5640)
                     /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(4998..5017)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5811..6399
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6300..6319
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6553..6570
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6638..6772
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6662..6681)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6716..6735
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6778..6973)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6792..6927
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7032..7053
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7068..7073,7074..7091,7092..7098)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7106..7122
                     /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     7111..7133
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7130..7146
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7130..7146
                     /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 ggcaggcgca acgctgcgac gaagacctcc
      601 gtttaagttc ttagccttcg ggggagaagc ggaacgcgcc aaagggcgag gccccggacc
      661 cgcctctgac cacgctgcat caccacctgg cgatggactt gccccagggg ctgctgctgc
      721 acgtccaccc tcacgcgacg cacgtgcgga cgctttattc gcatcgctca aagctggact
      781 cttgggaaaa aaggagtacc gggaagcggc cgcacttccc ccggagcctc cgcttccgcc
      841 cttgtacagc tcgtccatgc cgagagtgat cccggcggcg gtcacgaact ccagcaggac
      901 catgtgatcg cgcttctcgt tggggtcttt gctcagcttg gactgggtgc tcaggtagtg
      961 gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc
     1021 gagctgcacg ctgccgtcct cgatgttgtg gcggatcttg aagttcacct tgatgccgtt
     1081 cttctgcttg tcggccatga tatagacgtt gtggctgttg tagttgtact ccagcttgtg
     1141 ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc ggttcaccag
     1201 ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa
     1261 gatggtgcgc tcctggacgt agccttcggg catggcggac ttgaagaagt cgtgctgctt
     1321 catgtggtcg gggtagcggc tgaagcactg cacgccgtag gtcagggtgg tcacgagggt
     1381 gggccagggc acgggcagct tgccggtggt gcagatgaac ttcagggtca gcttgccgta
     1441 ggtggcatcg ccctcgccct cgccggacac gctgaacttg tggccgttta cgtcgccgtc
     1501 cagctcgacc aggatgggca ccaccccggt gaacagctcc tcgcccttgc tcaccatggt
     1561 ggcgaccggt ggatcccggg cccgcggtac cgtcgactgc agaattcggt tcactaaacg
     1621 agcccggttc actaaacgag ctctgcttat atagacctcc caccgtacac gcctaccgcc
     1681 catttgcgtc aatggggcgg agttgttacg acattttgga aagtcccgtt gattttggtg
     1741 ccaaaacaaa ctcccattga cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa
     1801 ccgctatcca cgcccattga tgtactgcca aaaccgcatc accatggtaa tagcgatgac
     1861 taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg
     1921 ccaggcgggc catttaccgt cattgacgtc aatagggggc gtacttggca tatgatacac
     1981 ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa
     2041 agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc
     2101 gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacgcgga actccatata
     2161 tgggctatga actaatgacc ccgtaattga ttactattaa taactagcac gcgttcgaag
     2221 cttgagctcg gtcccctcgg ggttgggagg tgggtctgaa acgataatgg tgaatatccc
     2281 tgcctaactc tattcactat agaaagtaca gcaaaaacta ttcttaaacc taccaagcct
     2341 cctactatca ttatgaataa ttttatatac cacagccaat ttgttatgtt aaaccaattc
     2401 cacaaacttg cccatttatc taattccaat aattcttgtt cattcttttc ttgctggttt
     2461 tgcgattctt caattaagga gtgtattaag cttgtgtaat tgttaatttc tctgtcccac
     2521 tccatccagg tcgtgtgatt ccaaatctgt tccagagatt tattactcca actagcattc
     2581 caaggcacag cagtggtgca aatgagtttt ccagagcaac cccaaatccc caggagctgt
     2641 tgatccttta ggtatctttc cacagccagg attcttgcct ggagctgctt gatgccccag
     2701 actgtgagtt gcaacagatg ctgttgcgcc tcaatagccc tcagcaaatt gttctgctgc
     2761 tgcactatac cagacaataa ttgtctggcc tgtaccgtca gcgtcattga cgctgcgccc
     2821 atagtgcttc ctgctgctcc caagaaccca aggaacaaag ctcctattcc cactgctctt
     2881 ttttctctct gcaccactct tctctttgcc ttggtgggtg ctactcctaa tggttcaatt
     2941 tttactactt tatatttata taattcactt ctccaattgt ccctcatatc tcctccttat
     3001 ggtgttttac taaacttttc catgttctaa tcctcatcct gtctacttgc cacacaatca
     3061 tcacctgcca tctgttttcc ataatcccta atgatctttg cttttcttct tggcactact
     3121 tttatgtcac tattatcttg tattactact gccccttcac ctttccagag gagctttgct
     3181 ggtcctttcc aaagtggatt tctgctgtcc ctgtaataaa cccgaaaatt ttgaattttt
     3241 gtaatttgtt tttgtaattc tttagtttgt atgtctgttg ctattatgtc tactattctt
     3301 tcccctgcac tgtacccccc aatcccccct tttcttttaa aattgtggat gaatactgcc
     3361 atttgtactg ctgtcttaag atgttcagcc tgatctctta cctgtcctat aattttcttt
     3421 aattctttat tcatagattc tactactcct tgactttggg gattgtaggg aatttccagg
     3481 tctgaagatc ctgctgcttg ctgtgctttt ttcttacttt tgttttgctc ttcctctatc
     3541 ttgtctaaag cttccttggt gtcttttatc tctatccttt gatgcacaca atagagggtt
     3601 gctactgtat tatataatga tctaagttct tctgatcctg tctgaaggga tggttgtagc
     3661 tgtcccagta tttgtctaca gccttctgat gtttctaaca ggccagtatt aactgcgaat
     3721 cgttctagct ccctgcttgc ccatactata tgttttaatt tatatttttt ctttccccct
     3781 ggccttaacc gaattttttc ccatcgcgat ctaattctcc cccgcttaat actgacgctc
     3841 tcgcacccat ctctctcctt ctagcctccg ctagtcaaaa tttttggcgt actcaccagt
     3901 cgccgcccct cgcctcttgc cgtgcgcgct tcagcaagcc gagtcctgcg tcgagagagc
     3961 tcctctggtt tccctttcgc tttcaagtcc ctgttcgggc gccactgcta gagattttcc
     4021 acactgacta aaagggtctg agggatctct agttaccaga gtcacacaac agacgggcac
     4081 acactacttg aagcactcaa ggcaagcttt attgaggctt aagcagtggg ttccctagtt
     4141 agccagagag ctcccaggct cagatctggt ctaaccagag agacccagta caggcaaaaa
     4201 gcagctgctt atatgcagga tctgagggct cgccactccc cagtcccgcc caggccacgc
     4261 ctcccatggt aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccata
     4321 aggtcatgta ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg
     4381 gcgtacttgg catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact
     4441 ccacccattg acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta
     4501 ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg cgggccattt accgtaagtt
     4561 atgtaacgcg gaactccata tatgggctat gaactaatga ccccgtaatt gattactatt
     4621 aataactagt caataatcaa tgtcgaccca ggtggcactt ttcggggaaa tgtgcgcgga
     4681 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4741 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     4801 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4861 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4921 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4981 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     5041 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     5101 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     5161 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     5221 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5281 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     5341 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5401 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5461 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5521 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5581 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5641 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     5701 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5761 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5821 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5881 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5941 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     6001 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6061 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6121 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6181 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6241 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6301 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6361 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6421 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     6481 gcttctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     6541 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     6601 cctctccccg cgcgttggcc gattcattaa tgcagcggat ccagacatga taagatacat
     6661 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     6721 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttagcttttt
     6781 gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggccga
     6841 ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc
     6901 ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga
     6961 ctaattgaga tgcggatccg ctggcacgac aggtttcccg actggaaagc gggcagtgag
     7021 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     7081 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     7141 tatgaccatg attacgccaa gcttgggctg caggtcgagg gatctccata agagaagagg
     7201 gacagctatg actgggagta gtcaggagag gaggaaaaat ctggctagta aaacatgtaa
     7261 ggaaaatttt agggatgtta aagaaaaaaa taacacaaaa caaaatat
//
