LOCUS       sequence-487962-        7566 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_049, derived from human fusion protein DCAF15_ZNF333.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7566)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7566)
  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_049
FEATURES             Location/Qualifiers
     source          1..7566
                     /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(1099..1809,1810..1812,1813..1815))
                     /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(1141..1162)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1490..1509
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1749..1770
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1812..1821
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1897..2100)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1926..1946)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2101..2404
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2661..2705)
                     /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    2890..3123
                     /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    3483..3600
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4092..4217
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4264..4521
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4523..4902
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4933..5037
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5038..5106,5107..5898)
                     /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(5256..5275)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6069..6657
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6558..6577
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6811..6828
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6896..7030
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6920..6939)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6974..6993
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7036..7231)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7050..7185
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7290..7311
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7326..7331,7332..7349,7350..7356)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7364..7380
                     /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     7369..7391
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7388..7404
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7388..7404
                     /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 aggctcctgc gaacatgctg ttggcagacc
      601 cggtgccata gcttcctctt caactttatc ttcttgtaac cacgctggat cgcgtgcggg
      661 gaccgcagag tcacgcgcca cccacggatt gcggtgacca agcgttggta ctggaataac
      721 gacgcgttgg atctgcatcg cggccttcaa acccgtacaa cccagtgacc acggaggctc
      781 ctgaagggct gggcggtcct cgcgcgtcag cggagtctcg cgttcctcga tagagggtac
      841 aagttgcatg cgtaagaaca acccaggccc catctgcatg tcacgggagg aggcctcctc
      901 aagcaggtcc tgcatggacg gcagttcttc aggcttcagt tgagattccc acgccacgcc
      961 cgttgcacgt aaaatgccgc gctccgttgc tttcggttca gcgcgttgtc ccagttgact
     1021 tacgcaactg ggtttacaag gtggagtccc tgccgaccgg gaagcggccg cacttccccc
     1081 ggagcctccg cttccgccct tgtacagctc gtccatgccg agagtgatcc cggcggcggt
     1141 cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc tcagcttgga
     1201 ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt
     1261 ctgctggtag tggtcggcga gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa
     1321 gttcaccttg atgccgttct tctgcttgtc ggccatgata tagacgttgt ggctgttgta
     1381 gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag
     1441 ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt
     1501 gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt
     1561 gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg aagcactgca cgccgtaggt
     1621 cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatgaactt
     1681 cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg
     1741 gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc
     1801 gcccttgctc accatggtgg cgaccggtgg atcccgggcc cgcggtaccg tcgactgcag
     1861 aattcggttc actaaacgag cccggttcac taaacgagct ctgcttatat agacctccca
     1921 ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag ttgttacgac attttggaaa
     1981 gtcccgttga ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg
     2041 aaatccccgt gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcac
     2101 catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt
     2161 catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt
     2221 acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac
     2281 ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga
     2341 cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt
     2401 aacgcggaac tccatatatg ggctatgaac taatgacccc gtaattgatt actattaata
     2461 actagcacgc gttcgaagct tgagctcggt cccctcgggg ttgggaggtg ggtctgaaac
     2521 gataatggtg aatatccctg cctaactcta ttcactatag aaagtacagc aaaaactatt
     2581 cttaaaccta ccaagcctcc tactatcatt atgaataatt ttatatacca cagccaattt
     2641 gttatgttaa accaattcca caaacttgcc catttatcta attccaataa ttcttgttca
     2701 ttcttttctt gctggttttg cgattcttca attaaggagt gtattaagct tgtgtaattg
     2761 ttaatttctc tgtcccactc catccaggtc gtgtgattcc aaatctgttc cagagattta
     2821 ttactccaac tagcattcca aggcacagca gtggtgcaaa tgagttttcc agagcaaccc
     2881 caaatcccca ggagctgttg atcctttagg tatctttcca cagccaggat tcttgcctgg
     2941 agctgcttga tgccccagac tgtgagttgc aacagatgct gttgcgcctc aatagccctc
     3001 agcaaattgt tctgctgctg cactatacca gacaataatt gtctggcctg taccgtcagc
     3061 gtcattgacg ctgcgcccat agtgcttcct gctgctccca agaacccaag gaacaaagct
     3121 cctattccca ctgctctttt ttctctctgc accactcttc tctttgcctt ggtgggtgct
     3181 actcctaatg gttcaatttt tactacttta tatttatata attcacttct ccaattgtcc
     3241 ctcatatctc ctccttatgg tgttttacta aacttttcca tgttctaatc ctcatcctgt
     3301 ctacttgcca cacaatcatc acctgccatc tgttttccat aatccctaat gatctttgct
     3361 tttcttcttg gcactacttt tatgtcacta ttatcttgta ttactactgc cccttcacct
     3421 ttccagagga gctttgctgg tcctttccaa agtggatttc tgctgtccct gtaataaacc
     3481 cgaaaatttt gaatttttgt aatttgtttt tgtaattctt tagtttgtat gtctgttgct
     3541 attatgtcta ctattctttc ccctgcactg taccccccaa tccccccttt tcttttaaaa
     3601 ttgtggatga atactgccat ttgtactgct gtcttaagat gttcagcctg atctcttacc
     3661 tgtcctataa ttttctttaa ttctttattc atagattcta ctactccttg actttgggga
     3721 ttgtagggaa tttccaggtc tgaagatcct gctgcttgct gtgctttttt cttacttttg
     3781 ttttgctctt cctctatctt gtctaaagct tccttggtgt cttttatctc tatcctttga
     3841 tgcacacaat agagggttgc tactgtatta tataatgatc taagttcttc tgatcctgtc
     3901 tgaagggatg gttgtagctg tcccagtatt tgtctacagc cttctgatgt ttctaacagg
     3961 ccagtattaa ctgcgaatcg ttctagctcc ctgcttgccc atactatatg ttttaattta
     4021 tattttttct ttccccctgg ccttaaccga attttttccc atcgcgatct aattctcccc
     4081 cgcttaatac tgacgctctc gcacccatct ctctccttct agcctccgct agtcaaaatt
     4141 tttggcgtac tcaccagtcg ccgcccctcg cctcttgccg tgcgcgcttc agcaagccga
     4201 gtcctgcgtc gagagagctc ctctggtttc cctttcgctt tcaagtccct gttcgggcgc
     4261 cactgctaga gattttccac actgactaaa agggtctgag ggatctctag ttaccagagt
     4321 cacacaacag acgggcacac actacttgaa gcactcaagg caagctttat tgaggcttaa
     4381 gcagtgggtt ccctagttag ccagagagct cccaggctca gatctggtct aaccagagag
     4441 acccagtaca ggcaaaaagc agctgcttat atgcaggatc tgagggctcg ccactcccca
     4501 gtcccgccca ggccacgcct cccatggtaa tagcgatgac taatacgtag atgtactgcc
     4561 aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt
     4621 cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg
     4681 cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta
     4741 tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg
     4801 ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc
     4861 ccgtaattga ttactattaa taactagtca ataatcaatg tcgacccagg tggcactttt
     4921 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4981 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5041 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5101 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5161 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5221 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5281 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5341 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5401 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5461 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5521 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5581 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5641 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5701 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5761 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5821 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5881 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5941 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6001 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6061 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6121 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6181 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     6241 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6301 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6361 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6421 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6481 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6541 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6601 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6661 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     6721 tttcctgcgt tatcccctgc ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     6781 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     6841 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagcggatcc
     6901 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     6961 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     7021 taaacaagtt agctttttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg
     7081 gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc
     7141 atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg
     7201 cgggactatg gttgctgact aattgagatg cggatccgct ggcacgacag gtttcccgac
     7261 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     7321 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     7381 tttcacacag gaaacagcta tgaccatgat tacgccaagc ttgggctgca ggtcgaggga
     7441 tctccataag agaagaggga cagctatgac tgggagtagt caggagagga ggaaaaatct
     7501 ggctagtaaa acatgtaagg aaaattttag ggatgttaaa gaaaaaaata acacaaaaca
     7561 aaatat
//
