LOCUS       sequence-487984-        8475 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_075, derived from human fusion protein JAK2_TPR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8475)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8475)
  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_075
FEATURES             Location/Qualifiers
     source          1..8475
                     /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(2008..2718,2719..2721,2722..2724))
                     /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(2050..2071)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2399..2418
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2658..2679
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2721..2730
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2806..3009)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2835..2855)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3010..3313
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3570..3614)
                     /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    3799..4032
                     /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    4392..4509
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5001..5126
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5173..5430
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5432..5811
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5842..5946
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5947..6015,6016..6807)
                     /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(6165..6184)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6978..7566
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7467..7486
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7720..7737
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7805..7939
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7829..7848)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7883..7902
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7945..8140)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7959..8094
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    8199..8220
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8235..8240,8241..8258,8259..8265)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8273..8289
                     /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     8278..8300
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8297..8313
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8297..8313
                     /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 gttaatatta cctctgttaa taccacctct
      601 gccgcccatg gcgtgagata cgcccctttg tcggttaaac tgtctacccc tgacaccttg
      661 gcgaagagta gtttgcagcc gaactctcct gaaaggcttt ggctgactag agcttgtgtc
      721 tacactgcta gaactcggtg ggtcctggct agtagaaggg agatctgact catcgctagc
      781 ttgcaccggc tcctcttcct gttgagagtc gatctcaagg ccggcctcag atgaaatacc
      841 ctcggattcg gcttccacga aaacctcatc tccatcgtcg cctgttgcgg tttcgtttgt
      901 agtgctcaag gtgccagtag aagtcgtcac cattgggacg gactgacttg cgtgctcgct
      961 agcatctgag gtagtgcttt ctgtaaacac tgtcacaggt gcggctacct ggaggggcgt
     1021 tgttgggacg cttcgaccac cactctcctc ttcatgcgcg aggaaaaggg gagtctcgta
     1081 cataccgagt ccgccttgag aggcgagttg gccgagatcg ctatgggaac tactcgtttg
     1141 aggcatgtcc tcaggaggac caaagcggaa tcgaggaaca cccgctactt gaggactatg
     1201 tatagcttct gcaaagccat cagtcctatg aggcaccaca agggtaggcg tggatggtac
     1261 tgtcctgtcc tcatcatcga aaaaatgttg ctgcatacca cctatcccag gggtcaactg
     1321 gaggcctctg ccaactgact gtcgccgagt catctgtatg cgttgaaccg gaggacccag
     1381 ctcctgtgga ggcgcgtgaa tggtaaggcg cggtgggaga ggatgggggg gtctccgtgg
     1441 actttgggga gccctcggag cttgtctttc ggaggctgag cttggctgct gctccctgct
     1501 cacttcctgt gaaaagctac tctcagctgc gccggtattc ccctccccag aattttggct
     1561 gtccgcggct ctgtgattcc cctctccacc acccatggat tcctcggtct ctgtgccagg
     1621 gtcagttccg tcccccccct cagcgtcatc ggcttcgtac ccgtcgttgc catcggcgct
     1681 acccgtccct tcattactat cttcaccttc atcccccatg ccagtgtcgt cctcatcgtc
     1741 atcatcgtcc tcttcatcct cctcataatc ttcatgttcg ccgtcatttt cgtcgtcatc
     1801 ctcttcttca tcatctgaat ctattacaat cacatcatcg cccttacctt gtccatcctg
     1861 agaacttgta gtagtttgct ggtcactttg aagggggccg agatcgattt gcagagattg
     1921 gctagtctct tctgaatcct ccataggcgt gtaatctcct tgcgtccggg aagcggccgc
     1981 acttcccccg gagcctccgc ttccgccctt gtacagctcg tccatgccga gagtgatccc
     2041 ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct
     2101 cagcttggac tgggtgctca ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat
     2161 gggggtgttc tgctggtagt ggtcggcgag ctgcacgctg ccgtcctcga tgttgtggcg
     2221 gatcttgaag ttcaccttga tgccgttctt ctgcttgtcg gccatgatat agacgttgtg
     2281 gctgttgtag ttgtactcca gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat
     2341 gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg aacttcacct cggcgcgggt
     2401 cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat
     2461 ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg tagcggctga agcactgcac
     2521 gccgtaggtc agggtggtca cgagggtggg ccagggcacg ggcagcttgc cggtggtgca
     2581 gatgaacttc agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct
     2641 gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg atgggcacca ccccggtgaa
     2701 cagctcctcg cccttgctca ccatggtggc gaccggtgga tcccgggccc gcggtaccgt
     2761 cgactgcaga attcggttca ctaaacgagc ccggttcact aaacgagctc tgcttatata
     2821 gacctcccac cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt tgttacgaca
     2881 ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg
     2941 gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattgatgt actgccaaaa
     3001 ccgcatcacc atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc
     3061 ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat
     3121 agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa
     3181 atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt
     3241 cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt
     3301 aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta
     3361 ctattaataa ctagcacgcg ttcgaagctt gagctcggtc ccctcggggt tgggaggtgg
     3421 gtctgaaacg ataatggtga atatccctgc ctaactctat tcactataga aagtacagca
     3481 aaaactattc ttaaacctac caagcctcct actatcatta tgaataattt tatataccac
     3541 agccaatttg ttatgttaaa ccaattccac aaacttgccc atttatctaa ttccaataat
     3601 tcttgttcat tcttttcttg ctggttttgc gattcttcaa ttaaggagtg tattaagctt
     3661 gtgtaattgt taatttctct gtcccactcc atccaggtcg tgtgattcca aatctgttcc
     3721 agagatttat tactccaact agcattccaa ggcacagcag tggtgcaaat gagttttcca
     3781 gagcaacccc aaatccccag gagctgttga tcctttaggt atctttccac agccaggatt
     3841 cttgcctgga gctgcttgat gccccagact gtgagttgca acagatgctg ttgcgcctca
     3901 atagccctca gcaaattgtt ctgctgctgc actataccag acaataattg tctggcctgt
     3961 accgtcagcg tcattgacgc tgcgcccata gtgcttcctg ctgctcccaa gaacccaagg
     4021 aacaaagctc ctattcccac tgctcttttt tctctctgca ccactcttct ctttgccttg
     4081 gtgggtgcta ctcctaatgg ttcaattttt actactttat atttatataa ttcacttctc
     4141 caattgtccc tcatatctcc tccttatggt gttttactaa acttttccat gttctaatcc
     4201 tcatcctgtc tacttgccac acaatcatca cctgccatct gttttccata atccctaatg
     4261 atctttgctt ttcttcttgg cactactttt atgtcactat tatcttgtat tactactgcc
     4321 ccttcacctt tccagaggag ctttgctggt cctttccaaa gtggatttct gctgtccctg
     4381 taataaaccc gaaaattttg aatttttgta atttgttttt gtaattcttt agtttgtatg
     4441 tctgttgcta ttatgtctac tattctttcc cctgcactgt accccccaat cccccctttt
     4501 cttttaaaat tgtggatgaa tactgccatt tgtactgctg tcttaagatg ttcagcctga
     4561 tctcttacct gtcctataat tttctttaat tctttattca tagattctac tactccttga
     4621 ctttggggat tgtagggaat ttccaggtct gaagatcctg ctgcttgctg tgcttttttc
     4681 ttacttttgt tttgctcttc ctctatcttg tctaaagctt ccttggtgtc ttttatctct
     4741 atcctttgat gcacacaata gagggttgct actgtattat ataatgatct aagttcttct
     4801 gatcctgtct gaagggatgg ttgtagctgt cccagtattt gtctacagcc ttctgatgtt
     4861 tctaacaggc cagtattaac tgcgaatcgt tctagctccc tgcttgccca tactatatgt
     4921 tttaatttat attttttctt tccccctggc cttaaccgaa ttttttccca tcgcgatcta
     4981 attctccccc gcttaatact gacgctctcg cacccatctc tctccttcta gcctccgcta
     5041 gtcaaaattt ttggcgtact caccagtcgc cgcccctcgc ctcttgccgt gcgcgcttca
     5101 gcaagccgag tcctgcgtcg agagagctcc tctggtttcc ctttcgcttt caagtccctg
     5161 ttcgggcgcc actgctagag attttccaca ctgactaaaa gggtctgagg gatctctagt
     5221 taccagagtc acacaacaga cgggcacaca ctacttgaag cactcaaggc aagctttatt
     5281 gaggcttaag cagtgggttc cctagttagc cagagagctc ccaggctcag atctggtcta
     5341 accagagaga cccagtacag gcaaaaagca gctgcttata tgcaggatct gagggctcgc
     5401 cactccccag tcccgcccag gccacgcctc ccatggtaat agcgatgact aatacgtaga
     5461 tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc
     5521 atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg
     5581 ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg
     5641 gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc
     5701 agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa
     5761 ctaatgaccc cgtaattgat tactattaat aactagtcaa taatcaatgt cgacccaggt
     5821 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     5881 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     5941 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     6001 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     6061 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     6121 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     6181 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     6241 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     6301 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     6361 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     6421 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     6481 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     6541 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     6601 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     6661 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     6721 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     6781 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     6841 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     6901 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     6961 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     7021 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     7081 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     7141 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     7201 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     7261 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     7321 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     7381 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     7441 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     7501 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     7561 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     7621 cacatgttct ttcctgcgtt atcccctgct tctgtggata accgtattac cgcctttgag
     7681 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     7741 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     7801 agcggatcca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     7861 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     7921 aagctgcaat aaacaagtta gctttttgca aaagcctagg cctccaaaaa agcctcctca
     7981 ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat
     8041 tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg
     8101 agttaggggc gggactatgg ttgctgacta attgagatgc ggatccgctg gcacgacagg
     8161 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     8221 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     8281 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct tgggctgcag
     8341 gtcgagggat ctccataaga gaagagggac agctatgact gggagtagtc aggagaggag
     8401 gaaaaatctg gctagtaaaa catgtaagga aaattttagg gatgttaaag aaaaaaataa
     8461 cacaaaacaa aatat
//
