LOCUS       sequence-488255-        8187 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            Vidr_026, derived from human protein SRSF4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8187)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8187)
  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_Vidr_026
FEATURES             Location/Qualifiers
     source          1..8187
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    828..962
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(852..871)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     906..925
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(968..1163)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      982..1117
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    1222..1243
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1258..1263,1264..1281,1282..1288)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1296..1312
                     /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     1301..1323
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1320..1336
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1320..1336
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1679..1698
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    1697..1752
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     1798..1817
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(join(3218..3928,3929..3931,3932..3934))
                     /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(3260..3281)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3609..3628
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3868..3889
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3931..3940
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4016..4219)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4045..4065)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4220..4523
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4780..4824)
                     /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    5009..5242
                     /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    5602..5719
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6211..6336
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6383..6640
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6642..7021
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7052..7156
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7157..7225,7226..8017)
                     /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(7375..7394)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagcggat ccagacatga
      841 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
      901 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
      961 ttagcttttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct
     1021 cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg
     1081 gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta
     1141 tggttgctga ctaattgaga tgcggatccg ctggcacgac aggtttcccg actggaaagc
     1201 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     1261 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     1321 aggaaacagc tatgaccatg attacgccaa gcttgggctg caggtcgagg gatctccata
     1381 agagaagagg gacagctatg actgggagta gtcaggagag gaggaaaaat ctggctagta
     1441 aaacatgtaa ggaaaatttt agggatgtta aagaaaaaaa taacacaaaa caaaatataa
     1501 aaaaaatcta acctcaagtc aaggcttttc tatggaataa ggaatggaca gcagggggct
     1561 gtttcatata ctgatgacct ctttatagcc aacctttgtt catggcagcc agcatatggg
     1621 catatgttgc caaactctaa accaaatact cattctgatg ttttaaatga tttgccctcc
     1681 catatgtcct tccgagtgag agacacaaaa aattccaaca cactattgca atgaaaataa
     1741 atttccttta ttagccagaa gtcagatgct caaggggctt catgatgtcc ccataatttt
     1801 tggcagaggg aaaaagatct gctagctcga gcagctgaag ctttattgag gcttaagcag
     1861 tgggttccct agttagccag agagctccca ggctcagatc tggtctaacc agagagaccc
     1921 agtacaggca aaaagcagat cttgtcttct ttgggagtga attagccctt ccagtccccc
     1981 cttttctttt aaaaagtggc taagatctac agctgccttg taagtcattg gtcttaaagg
     2041 taccgtaatc ttatctagaa ccggttcaac tacgcgagtg gctacggcta cgcgagcgag
     2101 aaggtgagcg ggaggcgcta cgagaacgag attttgagcg cgaacgggtc ttacttgccg
     2161 attttgaacg ggagcgagat tcggagggta aatttggttt actcttcgaa tttgagcgac
     2221 tgcgggagcg ggtctcttgg ttggtgcctg cgttctcgga ctcaccgcgt ccttcgcgct
     2281 gagaagactc agactttgca tgctcgcgct ccttgctcac tgaacgagat gggctgcgcg
     2341 actgggaacg atcggtatct tctttttttt tcttttttga cgacccagcc ttagaatcac
     2401 gtttcgagcc acgcttgcgg gagcgttcac tcttactgcg actacgacta cgcgaacggg
     2461 attcttcacg actgcgctta cggcccttac gtttatcctt tgacttagaa cggctacgag
     2521 agcgtgaacg ggacccgccc ttgctgcgac tacggctacg agactgacgc agggacttct
     2581 cctgtgaacg tccacgactg accgatccac gtttttcttc ctcgacgcgg cgctcctggg
     2641 agcgagagcg gctcttggac tttgacttgt gacgggatgg ggagcgcgac tttggcttac
     2701 ccacgttgtc attattctgg atcttttcct ctgcttggtc ttttgactta ctgcgtgatt
     2761 tgccagctga atgtgaacgg gagcgcgact tttccttact cggcgaacgt gatttttctt
     2821 ttttagagcg gctgcggctt tgtgaacgac tacgggattt tgaacgtgaa cgcgacccgc
     2881 tgcgcgagcg tgagcgagac ttactgtgtg acgacttgct agacccactg cgggaacgcg
     2941 atttgcgtga gtggcgcgag cgcgaacggg agcgactgtg actacggctg cgagaataag
     3001 agcgacgacg gcgtgagccc ggcttgtctt caaccaaacg gatcttgcga ccattcactt
     3061 ccgtgccatc taacttctca agcgcacgct tcatgtcact ataagagacg aactcgataa
     3121 caccctcgtt tttgcgccct ttgtgtgcat ctgcatacgt gacttcgcct gcctgccggg
     3181 aagcggccgc acttcccccg gagcctccgc ttccgccctt gtacagctcg tccatgccga
     3241 gagtgatccc ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg
     3301 ggtctttgct cagcttggac tgggtgctca ggtagtggtt gtcgggcagc agcacggggc
     3361 cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag ctgcacgctg ccgtcctcga
     3421 tgttgtggcg gatcttgaag ttcaccttga tgccgttctt ctgcttgtcg gccatgatat
     3481 agacgttgtg gctgttgtag ttgtactcca gcttgtgccc caggatgttg ccgtcctcct
     3541 tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg aacttcacct
     3601 cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc
     3661 cttcgggcat ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg tagcggctga
     3721 agcactgcac gccgtaggtc agggtggtca cgagggtggg ccagggcacg ggcagcttgc
     3781 cggtggtgca gatgaacttc agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc
     3841 cggacacgct gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg atgggcacca
     3901 ccccggtgaa cagctcctcg cccttgctca ccatggtggc gaccggtgga tcccgggccc
     3961 gcggtaccgt cgactgcaga attcggttca ctaaacgagc ccggttcact aaacgagctc
     4021 tgcttatata gacctcccac cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt
     4081 tgttacgaca ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt
     4141 caatggggtg gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattgatgt
     4201 actgccaaaa ccgcatcacc atggtaatag cgatgactaa tacgtagatg tactgccaag
     4261 taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat
     4321 tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag
     4381 tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg
     4441 gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc
     4501 catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg
     4561 taattgatta ctattaataa ctagcacgcg ttcgaagctt gagctcggtc ccctcggggt
     4621 tgggaggtgg gtctgaaacg ataatggtga atatccctgc ctaactctat tcactataga
     4681 aagtacagca aaaactattc ttaaacctac caagcctcct actatcatta tgaataattt
     4741 tatataccac agccaatttg ttatgttaaa ccaattccac aaacttgccc atttatctaa
     4801 ttccaataat tcttgttcat tcttttcttg ctggttttgc gattcttcaa ttaaggagtg
     4861 tattaagctt gtgtaattgt taatttctct gtcccactcc atccaggtcg tgtgattcca
     4921 aatctgttcc agagatttat tactccaact agcattccaa ggcacagcag tggtgcaaat
     4981 gagttttcca gagcaacccc aaatccccag gagctgttga tcctttaggt atctttccac
     5041 agccaggatt cttgcctgga gctgcttgat gccccagact gtgagttgca acagatgctg
     5101 ttgcgcctca atagccctca gcaaattgtt ctgctgctgc actataccag acaataattg
     5161 tctggcctgt accgtcagcg tcattgacgc tgcgcccata gtgcttcctg ctgctcccaa
     5221 gaacccaagg aacaaagctc ctattcccac tgctcttttt tctctctgca ccactcttct
     5281 ctttgccttg gtgggtgcta ctcctaatgg ttcaattttt actactttat atttatataa
     5341 ttcacttctc caattgtccc tcatatctcc tccttatggt gttttactaa acttttccat
     5401 gttctaatcc tcatcctgtc tacttgccac acaatcatca cctgccatct gttttccata
     5461 atccctaatg atctttgctt ttcttcttgg cactactttt atgtcactat tatcttgtat
     5521 tactactgcc ccttcacctt tccagaggag ctttgctggt cctttccaaa gtggatttct
     5581 gctgtccctg taataaaccc gaaaattttg aatttttgta atttgttttt gtaattcttt
     5641 agtttgtatg tctgttgcta ttatgtctac tattctttcc cctgcactgt accccccaat
     5701 cccccctttt cttttaaaat tgtggatgaa tactgccatt tgtactgctg tcttaagatg
     5761 ttcagcctga tctcttacct gtcctataat tttctttaat tctttattca tagattctac
     5821 tactccttga ctttggggat tgtagggaat ttccaggtct gaagatcctg ctgcttgctg
     5881 tgcttttttc ttacttttgt tttgctcttc ctctatcttg tctaaagctt ccttggtgtc
     5941 ttttatctct atcctttgat gcacacaata gagggttgct actgtattat ataatgatct
     6001 aagttcttct gatcctgtct gaagggatgg ttgtagctgt cccagtattt gtctacagcc
     6061 ttctgatgtt tctaacaggc cagtattaac tgcgaatcgt tctagctccc tgcttgccca
     6121 tactatatgt tttaatttat attttttctt tccccctggc cttaaccgaa ttttttccca
     6181 tcgcgatcta attctccccc gcttaatact gacgctctcg cacccatctc tctccttcta
     6241 gcctccgcta gtcaaaattt ttggcgtact caccagtcgc cgcccctcgc ctcttgccgt
     6301 gcgcgcttca gcaagccgag tcctgcgtcg agagagctcc tctggtttcc ctttcgcttt
     6361 caagtccctg ttcgggcgcc actgctagag attttccaca ctgactaaaa gggtctgagg
     6421 gatctctagt taccagagtc acacaacaga cgggcacaca ctacttgaag cactcaaggc
     6481 aagctttatt gaggcttaag cagtgggttc cctagttagc cagagagctc ccaggctcag
     6541 atctggtcta accagagaga cccagtacag gcaaaaagca gctgcttata tgcaggatct
     6601 gagggctcgc cactccccag tcccgcccag gccacgcctc ccatggtaat agcgatgact
     6661 aatacgtaga tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc
     6721 caggcgggcc atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact
     6781 tgatgtactg ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa
     6841 gtccctattg gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg
     6901 ttgggcggtc agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat
     6961 gggctatgaa ctaatgaccc cgtaattgat tactattaat aactagtcaa taatcaatgt
     7021 cgacccaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     7081 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     7141 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     7201 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     7261 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     7321 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     7381 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     7441 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     7501 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     7561 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     7621 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     7681 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     7741 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     7801 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     7861 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     7921 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     7981 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     8041 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     8101 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     8161 ccccgtagaa aagatcaaag gatcttc
//
