LOCUS       sequence-488168-        7536 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_147, derived from human fusion protein PAX5_ZCCHC7.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7536)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7536)
  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_147
FEATURES             Location/Qualifiers
     source          1..7536
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(81..100)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    146..201
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(200..219)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(562..578)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(562..578)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(575..597)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    586..602
                     /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)."
     promoter        complement(join(610..616,617..634,635..640))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    655..676
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        735..930
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      781..916
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    936..1070
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(973..992)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1027..1046
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1138..1155)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1309..1897)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1389..1408)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2068..2859,2860..2928))
                     /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     2691..2710
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2929..3033)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        3064..3443
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     LTR             3445..3702
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3749..3874
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4366..4483
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4843..5076
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5261..5305
                     /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"
     enhancer        5562..5865
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5866..6069
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6020..6040
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      6145..6154
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6151..6153,6154..6156,6157..6867)
                     /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(6196..6217)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6457..6476)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6804..6825
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
       61 cagctgctcg agctagcaga tctttttccc tctgccaaaa attatgggga catcatgaag
      121 ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt
      181 tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat
      241 cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac
      301 aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta
      361 ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt
      421 ttttctttaa catccctaaa attttcctta catgttttac tagccagatt tttcctcctc
      481 tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct cgacctgcag
      541 cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
      601 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
      661 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
      721 gtgccagcgg atccgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
      781 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
      841 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
      901 ggcttttttg gaggcctagg cttttgcaaa aagctaactt gtttattgca gcttataatg
      961 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     1021 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc cgctgcatta
     1081 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     1141 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     1201 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     1261 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1321 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1381 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1441 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1501 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1561 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1621 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     1681 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     1741 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     1801 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     1861 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     1921 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     1981 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     2041 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     2101 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     2161 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     2221 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     2281 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     2341 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     2401 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     2461 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     2521 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     2581 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     2641 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     2701 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     2761 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     2821 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     2881 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     2941 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     3001 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctgg
     3061 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
     3121 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     3181 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     3241 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     3301 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     3361 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     3421 tattagtcat cgctattacc atgggaggcg tggcctgggc gggactgggg agtggcgagc
     3481 cctcagatcc tgcatataag cagctgcttt ttgcctgtac tgggtctctc tggttagacc
     3541 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     3601 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     3661 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga
     3721 cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc
     3781 gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg
     3841 aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc
     3901 gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat
     3961 agtatgggca agcagggagc tagaacgatt cgcagttaat actggcctgt tagaaacatc
     4021 agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga
     4081 acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat
     4141 aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaaaaa
     4201 agcacagcaa gcagcaggat cttcagacct ggaaattccc tacaatcccc aaagtcaagg
     4261 agtagtagaa tctatgaata aagaattaaa gaaaattata ggacaggtaa gagatcaggc
     4321 tgaacatctt aagacagcag tacaaatggc agtattcatc cacaatttta aaagaaaagg
     4381 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     4441 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     4501 cagcagaaat ccactttgga aaggaccagc aaagctcctc tggaaaggtg aaggggcagt
     4561 agtaatacaa gataatagtg acataaaagt agtgccaaga agaaaagcaa agatcattag
     4621 ggattatgga aaacagatgg caggtgatga ttgtgtggca agtagacagg atgaggatta
     4681 gaacatggaa aagtttagta aaacaccata aggaggagat atgagggaca attggagaag
     4741 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     4801 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     4861 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     4921 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     4981 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     5041 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     5101 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     5161 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     5221 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     5281 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     5341 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     5401 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     5461 cccaaccccg aggggaccga gctcaagctt cgaacgcgtg ctagttatta atagtaatca
     5521 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
     5581 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     5641 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     5701 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     5761 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
     5821 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
     5881 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
     5941 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
     6001 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
     6061 agcagagctc gtttagtgaa ccgggctcgt ttagtgaacc gaattctgca gtcgacggta
     6121 ccgcgggccc gggatccacc ggtcgccacc atggtgagca agggcgagga gctgttcacc
     6181 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     6241 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     6301 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     6361 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     6421 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     6481 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     6541 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     6601 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     6661 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     6721 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccaa gctgagcaaa
     6781 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     6841 actctcggca tggacgagct gtacaagggc ggaagcggag gctccggggg aagtgcggcc
     6901 gcttcccgga agcgcttaaa gcaaaagatc aaggtattaa aaaaaaatgg agtgatcccc
     6961 gagccttcaa aactgcccta catcaaggcc gctaatgaaa accctcacca cgatatccgc
     7021 aagggccgtg cgagctggaa atctaaccgc tggccacaag agaacaagga gacgcaaaag
     7081 gagatgaaaa acaaaaaccg taactgggaa aaacatcgta aagccgatcg ccatcgcgaa
     7141 gtcgatgaag attttccgcg tggtcccaag acctattctt cccccggtag ctttaagacc
     7201 caaaagccat cgaagccctt tcatcgctct tcgcattatc atacaagtcg tgaggataaa
     7261 agccctaaag aaggcaaacg tggaaagcaa aaaaaaaaag aacgttgttg ggaggacgat
     7321 gataacgaca acttattcct gattaagcag cgcaaaaaaa agtcctgaac cggttctaga
     7381 taagattacg gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     7441 tttaaaagaa aaggggggac tggaagggct aattcactcc caaagaagac aagatctgct
     7501 ttttgcctgt actgggtctc tctggttaga ccagat
//
