LOCUS       sequence-488254-        8103 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            Vidr_025, derived from human protein NKAP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8103)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8103)
  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_025
FEATURES             Location/Qualifiers
     source          1..8103
                     /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(3134..3844,3845..3847,3848..3850))
                     /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(3176..3197)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3525..3544
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3784..3805
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3847..3856
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3932..4135)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3961..3981)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4136..4439
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4696..4740)
                     /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    4925..5158
                     /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    5518..5635
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6127..6252
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6299..6556
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6558..6937
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6968..7072
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7073..7141,7142..7933)
                     /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(7291..7310)
                     /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 ccggttcacg acgtaaggcc aatctcaccg cggcgtggga
     2101 tgcgcttccc ggctttcaca tattcagcca tcgctgcgcc ctcgccggga agcaatgcat
     2161 gtccgtagtt caatggtttg tcatcttggg atgttaatgt tttgggcgct tcaggcccga
     2221 taagatcact tggctcctcg gccttggtac gatctttcca tggattttcc agaaactctt
     2281 cttggctttc tttactgctt gaatcggacg attctttacg tgacttcttg ctacgctttt
     2341 ttttatattt tttcgagcga tgtttcttct ttttctcttt cttttttgcc ttcttcgcac
     2401 ggcgcttgtt atcctcgtca gacgagtcag tttctgagtc cgagtcagag tcgctatcct
     2461 cactgtattt tttgtgttta cgcttactac tttttttttt acgacgtttt ttagagcgtt
     2521 ctttgctgcg gctggatttc ttttttttct cttcctcgga ggtgctagcc gacgtggtac
     2581 tcttcttagg ttcctcgtcc tcgacagggg tatgctcgtc ggagtctggc tctgggttct
     2641 ttggacttaa cccccacact tctggggccc ccaattctcc gatgcgctca cgctcactta
     2701 aacgcttttg acgcaaagac tcctcacgct ccttgtccaa aagactcggc cacggcttat
     2761 ccgaaccata gggacgggag taagacccat aatagaccga gctactggca gatgcgaatg
     2821 gaataccacg tggtgcacta gggcgctcgc gagagcgtga acgtgagcgg gagcgataac
     2881 tctggttgcg tgagccttgc gacaggccgc ccaactgatg cgttaatcca ttgcgatcac
     2941 ctgaacggct gcaactatgg ctacgcgaac ggcggccacg ggggctacgc gcactcttgg
     3001 acggcttcgg agactttgat gaactgcgac ggcgacctcc cgaaccactc gcttcacgat
     3061 caggtgaacg cgacccgcta actggcgcca tccgggaagc ggccgcactt cccccggagc
     3121 ctccgcttcc gcccttgtac agctcgtcca tgccgagagt gatcccggcg gcggtcacga
     3181 actccagcag gaccatgtga tcgcgcttct cgttggggtc tttgctcagc ttggactggg
     3241 tgctcaggta gtggttgtcg ggcagcagca cggggccgtc gccgatgggg gtgttctgct
     3301 ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt gtggcggatc ttgaagttca
     3361 ccttgatgcc gttcttctgc ttgtcggcca tgatatagac gttgtggctg ttgtagttgt
     3421 actccagctt gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga
     3481 tgcggttcac cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt
     3541 cgtccttgaa gaagatggtg cgctcctgga cgtagccttc gggcatggcg gacttgaaga
     3601 agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca ctgcacgccg taggtcaggg
     3661 tggtcacgag ggtgggccag ggcacgggca gcttgccggt ggtgcagatg aacttcaggg
     3721 tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt
     3781 ttacgtcgcc gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct
     3841 tgctcaccat ggtggcgacc ggtggatccc gggcccgcgg taccgtcgac tgcagaattc
     3901 ggttcactaa acgagcccgg ttcactaaac gagctctgct tatatagacc tcccaccgta
     3961 cacgcctacc gcccatttgc gtcaatgggg cggagttgtt acgacatttt ggaaagtccc
     4021 gttgattttg gtgccaaaac aaactcccat tgacgtcaat ggggtggaga cttggaaatc
     4081 cccgtgagtc aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc atcaccatgg
     4141 taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt
     4201 actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg
     4261 gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt
     4321 gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca
     4381 atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc
     4441 ggaactccat atatgggcta tgaactaatg accccgtaat tgattactat taataactag
     4501 cacgcgttcg aagcttgagc tcggtcccct cggggttggg aggtgggtct gaaacgataa
     4561 tggtgaatat ccctgcctaa ctctattcac tatagaaagt acagcaaaaa ctattcttaa
     4621 acctaccaag cctcctacta tcattatgaa taattttata taccacagcc aatttgttat
     4681 gttaaaccaa ttccacaaac ttgcccattt atctaattcc aataattctt gttcattctt
     4741 ttcttgctgg ttttgcgatt cttcaattaa ggagtgtatt aagcttgtgt aattgttaat
     4801 ttctctgtcc cactccatcc aggtcgtgtg attccaaatc tgttccagag atttattact
     4861 ccaactagca ttccaaggca cagcagtggt gcaaatgagt tttccagagc aaccccaaat
     4921 ccccaggagc tgttgatcct ttaggtatct ttccacagcc aggattcttg cctggagctg
     4981 cttgatgccc cagactgtga gttgcaacag atgctgttgc gcctcaatag ccctcagcaa
     5041 attgttctgc tgctgcacta taccagacaa taattgtctg gcctgtaccg tcagcgtcat
     5101 tgacgctgcg cccatagtgc ttcctgctgc tcccaagaac ccaaggaaca aagctcctat
     5161 tcccactgct cttttttctc tctgcaccac tcttctcttt gccttggtgg gtgctactcc
     5221 taatggttca atttttacta ctttatattt atataattca cttctccaat tgtccctcat
     5281 atctcctcct tatggtgttt tactaaactt ttccatgttc taatcctcat cctgtctact
     5341 tgccacacaa tcatcacctg ccatctgttt tccataatcc ctaatgatct ttgcttttct
     5401 tcttggcact acttttatgt cactattatc ttgtattact actgcccctt cacctttcca
     5461 gaggagcttt gctggtcctt tccaaagtgg atttctgctg tccctgtaat aaacccgaaa
     5521 attttgaatt tttgtaattt gtttttgtaa ttctttagtt tgtatgtctg ttgctattat
     5581 gtctactatt ctttcccctg cactgtaccc cccaatcccc ccttttcttt taaaattgtg
     5641 gatgaatact gccatttgta ctgctgtctt aagatgttca gcctgatctc ttacctgtcc
     5701 tataattttc tttaattctt tattcataga ttctactact ccttgacttt ggggattgta
     5761 gggaatttcc aggtctgaag atcctgctgc ttgctgtgct tttttcttac ttttgttttg
     5821 ctcttcctct atcttgtcta aagcttcctt ggtgtctttt atctctatcc tttgatgcac
     5881 acaatagagg gttgctactg tattatataa tgatctaagt tcttctgatc ctgtctgaag
     5941 ggatggttgt agctgtccca gtatttgtct acagccttct gatgtttcta acaggccagt
     6001 attaactgcg aatcgttcta gctccctgct tgcccatact atatgtttta atttatattt
     6061 tttctttccc cctggcctta accgaatttt ttcccatcgc gatctaattc tcccccgctt
     6121 aatactgacg ctctcgcacc catctctctc cttctagcct ccgctagtca aaatttttgg
     6181 cgtactcacc agtcgccgcc cctcgcctct tgccgtgcgc gcttcagcaa gccgagtcct
     6241 gcgtcgagag agctcctctg gtttcccttt cgctttcaag tccctgttcg ggcgccactg
     6301 ctagagattt tccacactga ctaaaagggt ctgagggatc tctagttacc agagtcacac
     6361 aacagacggg cacacactac ttgaagcact caaggcaagc tttattgagg cttaagcagt
     6421 gggttcccta gttagccaga gagctcccag gctcagatct ggtctaacca gagagaccca
     6481 gtacaggcaa aaagcagctg cttatatgca ggatctgagg gctcgccact ccccagtccc
     6541 gcccaggcca cgcctcccat ggtaatagcg atgactaata cgtagatgta ctgccaagta
     6601 ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg
     6661 acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt
     6721 taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga
     6781 acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca
     6841 tttaccgtaa gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta
     6901 attgattact attaataact agtcaataat caatgtcgac ccaggtggca cttttcgggg
     6961 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     7021 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     7081 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     7141 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     7201 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     7261 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     7321 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     7381 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     7441 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     7501 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     7561 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     7621 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     7681 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     7741 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     7801 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     7861 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     7921 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     7981 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     8041 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     8101 ttc
//
