LOCUS       sequence-488140-        8565 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_117, derived from human fusion protein NUP98_HOXD13.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8565)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8565)
  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_117
FEATURES             Location/Qualifiers
     source          1..8565
                     /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(2098..2808,2809..2811,2812..2814))
                     /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(2140..2161)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2489..2508
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2748..2769
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2811..2820
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2896..3099)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2925..2945)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3100..3403
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3660..3704)
                     /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    3889..4122
                     /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    4482..4599
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5091..5216
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5263..5520
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5522..5901
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5932..6036
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6037..6105,6106..6897)
                     /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(6255..6274)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7068..7656
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7557..7576
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7810..7827
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7895..8029
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7919..7938)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7973..7992
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(8035..8230)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      8049..8184
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    8289..8310
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8325..8330,8331..8348,8349..8355)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8363..8379
                     /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     8368..8390
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8387..8403
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8387..8403
                     /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 atcgggctga ttcagtgcga cgtcgcctac
      601 tggggcctgc ggcgccccaa aacccagtgt agccgtcgtg gtattgaagc ccggagcccc
      661 aaatgcgccg gtgcctaatg gacccccaat cttgggctgg ttgtttccga ataaactggc
      721 ttgccccgcg ccaagggccg ttccgaaccc agcgccaaga cccgttccaa gagttccggg
      781 cgctggtttt gagccgaaga tagagttccc tgaggtgttt gtaccgaaac caaaattaga
      841 ggtatttgtg ttagttccca atgtcagtgt aggtttattc ccaaaaagtc ctcccgatgt
      901 tgttccaaag gacggtgcgg acgtcgtact tgaaccgaag gtagttaatt tattattacc
      961 aaacaacgtc gatccaactg cgccaaatcc ggtattcgtt tgcccgaaca ggccggttcc
     1021 ggtgccgaag gcggtgccag tcgaggtgtt cgttgccgtt ccgaaaagac ctcccggctg
     1081 cgaagcctgt gttactccga ataaacccat tgtattcgtt gacggctgac cgattgtgga
     1141 agtattgccg aatgaaaagc cggtgttctg agtggtggtt gcttggccaa acggtttgga
     1201 aaacaggctc gtggtttgtt gattttgttg gccaaacaaa cctcccggat tagtcccaaa
     1261 accagtagta ctggtgccga acgcggtctt attttgccca taggcaaagc cgctgttggt
     1321 ggtagaagaa ctgaaaaggc ctgttgcgga agaagttgcc ggtgaacttc caaataatcc
     1381 ggtagtcgta cccgccccca cttggttctg cggtcctttg cggtttgcct gatagtcttc
     1441 caaacgcaat tcttcaagac ttttgctctc gtattccttc attgccgtaa tgcactggtg
     1501 ttttgtacta atgtttgtac tcacgcccgc tttcaccatg gtatctgtgc ctgtaggggg
     1561 attaaatttg atagtagttc ctgttggggc agccgtaaat gaactcggtc cgaacagact
     1621 tccgctcgta cttccaaagg ggttactcgt tgtgttcgtt gtgccgaaaa ggccgccact
     1681 tgaggttgaa gtcccgaaat taccaaaccc ggtgggttta ttttgtgcaa aggcgttatt
     1741 ctgggagcta aacagggacg tcccagtgga agcagtccca aataacgtat tggctgtccc
     1801 ggtggaggtt ccaaacccaa atcctgtaga agtagatgta gcgggttgac taaagctgga
     1861 agttccaaac aatcctccgg gtttagtttg tgaattgcca aacaaccccc cagtgttgtt
     1921 agagcttccg aaggcactcg ttccaaacgc gccaccgctg gttgttccga accccgtatt
     1981 ctgcccgaac gtactggtcg tgccgaaacc cccagtaccg cccccgaaag gtgttccgaa
     2041 cgacttgtta aacatccggg aagcggccgc acttcccccg gagcctccgc ttccgccctt
     2101 gtacagctcg tccatgccga gagtgatccc ggcggcggtc acgaactcca gcaggaccat
     2161 gtgatcgcgc ttctcgttgg ggtctttgct cagcttggac tgggtgctca ggtagtggtt
     2221 gtcgggcagc agcacggggc cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag
     2281 ctgcacgctg ccgtcctcga tgttgtggcg gatcttgaag ttcaccttga tgccgttctt
     2341 ctgcttgtcg gccatgatat agacgttgtg gctgttgtag ttgtactcca gcttgtgccc
     2401 caggatgttg ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt
     2461 gtcgccctcg aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat
     2521 ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg aagaagtcgt gctgcttcat
     2581 gtggtcgggg tagcggctga agcactgcac gccgtaggtc agggtggtca cgagggtggg
     2641 ccagggcacg ggcagcttgc cggtggtgca gatgaacttc agggtcagct tgccgtaggt
     2701 ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg ccgtttacgt cgccgtccag
     2761 ctcgaccagg atgggcacca ccccggtgaa cagctcctcg cccttgctca ccatggtggc
     2821 gaccggtgga tcccgggccc gcggtaccgt cgactgcaga attcggttca ctaaacgagc
     2881 ccggttcact aaacgagctc tgcttatata gacctcccac cgtacacgcc taccgcccat
     2941 ttgcgtcaat ggggcggagt tgttacgaca ttttggaaag tcccgttgat tttggtgcca
     3001 aaacaaactc ccattgacgt caatggggtg gagacttgga aatccccgtg agtcaaaccg
     3061 ctatccacgc ccattgatgt actgccaaaa ccgcatcacc atggtaatag cgatgactaa
     3121 tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca
     3181 ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg
     3241 atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt
     3301 ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt
     3361 gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg
     3421 gctatgaact aatgaccccg taattgatta ctattaataa ctagcacgcg ttcgaagctt
     3481 gagctcggtc ccctcggggt tgggaggtgg gtctgaaacg ataatggtga atatccctgc
     3541 ctaactctat tcactataga aagtacagca aaaactattc ttaaacctac caagcctcct
     3601 actatcatta tgaataattt tatataccac agccaatttg ttatgttaaa ccaattccac
     3661 aaacttgccc atttatctaa ttccaataat tcttgttcat tcttttcttg ctggttttgc
     3721 gattcttcaa ttaaggagtg tattaagctt gtgtaattgt taatttctct gtcccactcc
     3781 atccaggtcg tgtgattcca aatctgttcc agagatttat tactccaact agcattccaa
     3841 ggcacagcag tggtgcaaat gagttttcca gagcaacccc aaatccccag gagctgttga
     3901 tcctttaggt atctttccac agccaggatt cttgcctgga gctgcttgat gccccagact
     3961 gtgagttgca acagatgctg ttgcgcctca atagccctca gcaaattgtt ctgctgctgc
     4021 actataccag acaataattg tctggcctgt accgtcagcg tcattgacgc tgcgcccata
     4081 gtgcttcctg ctgctcccaa gaacccaagg aacaaagctc ctattcccac tgctcttttt
     4141 tctctctgca ccactcttct ctttgccttg gtgggtgcta ctcctaatgg ttcaattttt
     4201 actactttat atttatataa ttcacttctc caattgtccc tcatatctcc tccttatggt
     4261 gttttactaa acttttccat gttctaatcc tcatcctgtc tacttgccac acaatcatca
     4321 cctgccatct gttttccata atccctaatg atctttgctt ttcttcttgg cactactttt
     4381 atgtcactat tatcttgtat tactactgcc ccttcacctt tccagaggag ctttgctggt
     4441 cctttccaaa gtggatttct gctgtccctg taataaaccc gaaaattttg aatttttgta
     4501 atttgttttt gtaattcttt agtttgtatg tctgttgcta ttatgtctac tattctttcc
     4561 cctgcactgt accccccaat cccccctttt cttttaaaat tgtggatgaa tactgccatt
     4621 tgtactgctg tcttaagatg ttcagcctga tctcttacct gtcctataat tttctttaat
     4681 tctttattca tagattctac tactccttga ctttggggat tgtagggaat ttccaggtct
     4741 gaagatcctg ctgcttgctg tgcttttttc ttacttttgt tttgctcttc ctctatcttg
     4801 tctaaagctt ccttggtgtc ttttatctct atcctttgat gcacacaata gagggttgct
     4861 actgtattat ataatgatct aagttcttct gatcctgtct gaagggatgg ttgtagctgt
     4921 cccagtattt gtctacagcc ttctgatgtt tctaacaggc cagtattaac tgcgaatcgt
     4981 tctagctccc tgcttgccca tactatatgt tttaatttat attttttctt tccccctggc
     5041 cttaaccgaa ttttttccca tcgcgatcta attctccccc gcttaatact gacgctctcg
     5101 cacccatctc tctccttcta gcctccgcta gtcaaaattt ttggcgtact caccagtcgc
     5161 cgcccctcgc ctcttgccgt gcgcgcttca gcaagccgag tcctgcgtcg agagagctcc
     5221 tctggtttcc ctttcgcttt caagtccctg ttcgggcgcc actgctagag attttccaca
     5281 ctgactaaaa gggtctgagg gatctctagt taccagagtc acacaacaga cgggcacaca
     5341 ctacttgaag cactcaaggc aagctttatt gaggcttaag cagtgggttc cctagttagc
     5401 cagagagctc ccaggctcag atctggtcta accagagaga cccagtacag gcaaaaagca
     5461 gctgcttata tgcaggatct gagggctcgc cactccccag tcccgcccag gccacgcctc
     5521 ccatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg
     5581 tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg
     5641 tacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca
     5701 cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg
     5761 acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg
     5821 taacgcggaa ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactattaat
     5881 aactagtcaa taatcaatgt cgacccaggt ggcacttttc ggggaaatgt gcgcggaacc
     5941 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     6001 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     6061 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     6121 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     6181 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     6241 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     6301 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     6361 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     6421 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     6481 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     6541 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     6601 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     6661 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     6721 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     6781 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6841 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     6901 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6961 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     7021 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     7081 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     7141 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     7201 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     7261 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     7321 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     7381 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     7441 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     7501 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     7561 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     7621 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     7681 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     7741 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     7801 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     7861 ctccccgcgc gttggccgat tcattaatgc agcggatcca gacatgataa gatacattga
     7921 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     7981 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta gctttttgca
     8041 aaagcctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc
     8101 ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga
     8161 actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta
     8221 attgagatgc ggatccgctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     8281 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     8341 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     8401 gaccatgatt acgccaagct tgggctgcag gtcgagggat ctccataaga gaagagggac
     8461 agctatgact gggagtagtc aggagaggag gaaaaatctg gctagtaaaa catgtaagga
     8521 aaattttagg gatgttaaag aaaaaaataa cacaaaacaa aatat
//
