LOCUS       sequence-488261-        8472 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            Vidr_032, derived from human protein F90AE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8472)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8472)
  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_032
FEATURES             Location/Qualifiers
     source          1..8472
                     /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(3503..4213,4214..4216,4217..4219))
                     /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(3545..3566)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3894..3913
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4153..4174
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4216..4225
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4301..4504)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4330..4350)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4505..4808
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(5065..5109)
                     /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    5294..5527
                     /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    5887..6004
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6496..6621
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6668..6925
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6927..7306
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7337..7441
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7442..7510,7511..8302)
                     /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(7660..7679)
                     /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 ccggttcatt ctaagtcaga gtctgagtct tcagaagaag
     2101 aggaaacctg aagatcctcg taaagaacgc tcgggggaac acgcacacat ggggcttctg
     2161 atttttcgga gacatggggc gattgtgcca aaaatgcgcc cggtttttcg ggcgaatgaa
     2221 agctgggggc ggccaacaaa ctggacgacc agcgcccgtt ctcaaggcga cgaaacagaa
     2281 cacgcaaggg ttgtgcccca tcatgggagg ctgcgggatg gtgcgacatc gtacaagcct
     2341 gcgcggtagg taagcaggga cgactatgag gtggctggcg atctactgac ggtacttgtg
     2401 cgggtgtgcg acgacccatt tgcggactgg tagacggtcc taactcagtc gcagcaggtg
     2461 gaggctgaag gttctcgggt gctcccaact caccgccttg aattgcagac tcaggaattt
     2521 gaaacgggcc caggcgaggt ttcttgggga agttcaagca tgcttggatt ggcgcttgtg
     2581 cagggcgttt tgcacccggt ccgaacgaca agttcgaacc caaacctaag ctatgggtag
     2641 ctgccggggg acaaggttgt gacgtaacag cagggcgctt gtcctgtgcc tgcggacgtg
     2701 ccgcttgaag caaaccatgg gttttggacg cggcttgcgg aacttcacga cagtctccct
     2761 ctggacgaga atgggttggc ttgactacca gcaaaggttc gcggccttgg tgacgaacag
     2821 ctggctgcgg catgtctgca gcggcacccg tttgacgctc tttgggacca agagatgaag
     2881 atgaagataa ggacgctttg cgaaggggag acagagaggc cagtactgag ccacgcccac
     2941 tcatttccgt ggccgaacgg tcagctaaca cggggtctaa acgggggcgc ttgctagttg
     3001 tatgcacagg catggggcca ctggctacgc gtaagtagtc acttggttcc gtggagccct
     3061 taccattcgg cagtggcttc tcgggtggct ttccactaaa catgtgaagt aatgctttgc
     3121 gctgcgggtc ctgctggcgt gggcgctcct ctttttctcc cttatctttg ttaagtgggc
     3181 ctggatttgc ttcaacacgc ggtttccacg gcttcaagtt ctctttacct tcttttttgc
     3241 ccagcgtagc aggaaccagc gccgccttcc agcatttcat tgggcaacga gtactacgcg
     3301 cagtgtgtcc aaaagctccg caattcttac attttaagcg ggggtcctcc tcatccggag
     3361 ggggtgcacg aggtcctacc ggcgcacggc gctgtttttg cagggtttgc gcgcggacta
     3421 agcgtttagc ccatgatgtc ggatcacggc gagccatcat ccgggaagcg gccgcacttc
     3481 ccccggagcc tccgcttccg cccttgtaca gctcgtccat gccgagagtg atcccggcgg
     3541 cggtcacgaa ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcagct
     3601 tggactgggt gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg
     3661 tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg tggcggatct
     3721 tgaagttcac cttgatgccg ttcttctgct tgtcggccat gatatagacg ttgtggctgt
     3781 tgtagttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct
     3841 tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt
     3901 agttgccgtc gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg
     3961 acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg gctgaagcac tgcacgccgt
     4021 aggtcagggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatga
     4081 acttcagggt cagcttgccg taggtggcat cgccctcgcc ctcgccggac acgctgaact
     4141 tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg gtgaacagct
     4201 cctcgccctt gctcaccatg gtggcgaccg gtggatcccg ggcccgcggt accgtcgact
     4261 gcagaattcg gttcactaaa cgagcccggt tcactaaacg agctctgctt atatagacct
     4321 cccaccgtac acgcctaccg cccatttgcg tcaatggggc ggagttgtta cgacattttg
     4381 gaaagtcccg ttgattttgg tgccaaaaca aactcccatt gacgtcaatg gggtggagac
     4441 ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt gatgtactgc caaaaccgca
     4501 tcaccatggt aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccata
     4561 aggtcatgta ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg
     4621 gcgtacttgg catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact
     4681 ccacccattg acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta
     4741 ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg cgggccattt accgtaagtt
     4801 atgtaacgcg gaactccata tatgggctat gaactaatga ccccgtaatt gattactatt
     4861 aataactagc acgcgttcga agcttgagct cggtcccctc ggggttggga ggtgggtctg
     4921 aaacgataat ggtgaatatc cctgcctaac tctattcact atagaaagta cagcaaaaac
     4981 tattcttaaa cctaccaagc ctcctactat cattatgaat aattttatat accacagcca
     5041 atttgttatg ttaaaccaat tccacaaact tgcccattta tctaattcca ataattcttg
     5101 ttcattcttt tcttgctggt tttgcgattc ttcaattaag gagtgtatta agcttgtgta
     5161 attgttaatt tctctgtccc actccatcca ggtcgtgtga ttccaaatct gttccagaga
     5221 tttattactc caactagcat tccaaggcac agcagtggtg caaatgagtt ttccagagca
     5281 accccaaatc cccaggagct gttgatcctt taggtatctt tccacagcca ggattcttgc
     5341 ctggagctgc ttgatgcccc agactgtgag ttgcaacaga tgctgttgcg cctcaatagc
     5401 cctcagcaaa ttgttctgct gctgcactat accagacaat aattgtctgg cctgtaccgt
     5461 cagcgtcatt gacgctgcgc ccatagtgct tcctgctgct cccaagaacc caaggaacaa
     5521 agctcctatt cccactgctc ttttttctct ctgcaccact cttctctttg ccttggtggg
     5581 tgctactcct aatggttcaa tttttactac tttatattta tataattcac ttctccaatt
     5641 gtccctcata tctcctcctt atggtgtttt actaaacttt tccatgttct aatcctcatc
     5701 ctgtctactt gccacacaat catcacctgc catctgtttt ccataatccc taatgatctt
     5761 tgcttttctt cttggcacta cttttatgtc actattatct tgtattacta ctgccccttc
     5821 acctttccag aggagctttg ctggtccttt ccaaagtgga tttctgctgt ccctgtaata
     5881 aacccgaaaa ttttgaattt ttgtaatttg tttttgtaat tctttagttt gtatgtctgt
     5941 tgctattatg tctactattc tttcccctgc actgtacccc ccaatccccc cttttctttt
     6001 aaaattgtgg atgaatactg ccatttgtac tgctgtctta agatgttcag cctgatctct
     6061 tacctgtcct ataattttct ttaattcttt attcatagat tctactactc cttgactttg
     6121 gggattgtag ggaatttcca ggtctgaaga tcctgctgct tgctgtgctt ttttcttact
     6181 tttgttttgc tcttcctcta tcttgtctaa agcttccttg gtgtctttta tctctatcct
     6241 ttgatgcaca caatagaggg ttgctactgt attatataat gatctaagtt cttctgatcc
     6301 tgtctgaagg gatggttgta gctgtcccag tatttgtcta cagccttctg atgtttctaa
     6361 caggccagta ttaactgcga atcgttctag ctccctgctt gcccatacta tatgttttaa
     6421 tttatatttt ttctttcccc ctggccttaa ccgaattttt tcccatcgcg atctaattct
     6481 cccccgctta atactgacgc tctcgcaccc atctctctcc ttctagcctc cgctagtcaa
     6541 aatttttggc gtactcacca gtcgccgccc ctcgcctctt gccgtgcgcg cttcagcaag
     6601 ccgagtcctg cgtcgagaga gctcctctgg tttccctttc gctttcaagt ccctgttcgg
     6661 gcgccactgc tagagatttt ccacactgac taaaagggtc tgagggatct ctagttacca
     6721 gagtcacaca acagacgggc acacactact tgaagcactc aaggcaagct ttattgaggc
     6781 ttaagcagtg ggttccctag ttagccagag agctcccagg ctcagatctg gtctaaccag
     6841 agagacccag tacaggcaaa aagcagctgc ttatatgcag gatctgaggg ctcgccactc
     6901 cccagtcccg cccaggccac gcctcccatg gtaatagcga tgactaatac gtagatgtac
     6961 tgccaagtag gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta
     7021 ccgtcattga cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag
     7081 tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt
     7141 actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca
     7201 ggcgggccat ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat
     7261 gaccccgtaa ttgattacta ttaataacta gtcaataatc aatgtcgacc caggtggcac
     7321 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     7381 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     7441 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     7501 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     7561 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     7621 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     7681 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     7741 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     7801 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     7861 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     7921 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     7981 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     8041 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     8101 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     8161 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     8221 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     8281 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     8341 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     8401 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     8461 caaaggatct tc
//
