LOCUS       sequence-488246-        7491 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            Vidr_016, derived from human protein KDM5D.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7491)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7491)
  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_016
FEATURES             Location/Qualifiers
     source          1..7491
                     /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(2522..3232,3233..3235,3236..3238))
                     /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(2564..2585)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2913..2932
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3172..3193
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3235..3244
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3320..3523)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3349..3369)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3524..3827
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4084..4128)
                     /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    4313..4546
                     /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    4906..5023
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5515..5640
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5687..5944
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5946..6325
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6356..6460
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6461..6529,6530..7321)
                     /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(6679..6698)
                     /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 ccggttcaca actgctgctc gtctgagtaa ctcaactgta
     2101 acagcggcat caggctgggg caggccgcgc ttgatccgct atccttatgc tgtaatgagt
     2161 tttggttgcc ttttaagaag ggggagtgat ccgtgctagg tgttaaaaac atgttttcgc
     2221 ggtcggcctt ctcttggtaa tgttcctctt cgcgccctac ttgcgacata ataccacttg
     2281 agcgcgcacg cttgctctgt aattcttgct gaaccaagtt ttctacatta cgaccttggt
     2341 caactttctg gcggcggcgg cgacgttcta atgcgcggga acgagtacgc gaaccctggt
     2401 gctcgaactt ctccaattcc agcagagtac ggatacgatc caaatcgggg ggttgaccgg
     2461 cttgaagtaa ctgccaaatc cgggaagcgg ccgcacttcc cccggagcct ccgcttccgc
     2521 ccttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga
     2581 ccatgtgatc gcgcttctcg ttggggtctt tgctcagctt ggactgggtg ctcaggtagt
     2641 ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg
     2701 cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc ttgatgccgt
     2761 tcttctgctt gtcggccatg atatagacgt tgtggctgtt gtagttgtac tccagcttgt
     2821 gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca
     2881 gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga
     2941 agatggtgcg ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct
     3001 tcatgtggtc ggggtagcgg ctgaagcact gcacgccgta ggtcagggtg gtcacgaggg
     3061 tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt
     3121 aggtggcatc gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt
     3181 ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatgg
     3241 tggcgaccgg tggatcccgg gcccgcggta ccgtcgactg cagaattcgg ttcactaaac
     3301 gagcccggtt cactaaacga gctctgctta tatagacctc ccaccgtaca cgcctaccgc
     3361 ccatttgcgt caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt
     3421 gccaaaacaa actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa
     3481 accgctatcc acgcccattg atgtactgcc aaaaccgcat caccatggta atagcgatga
     3541 ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat
     3601 gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca
     3661 cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga
     3721 aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt
     3781 cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat
     3841 atgggctatg aactaatgac cccgtaattg attactatta ataactagca cgcgttcgaa
     3901 gcttgagctc ggtcccctcg gggttgggag gtgggtctga aacgataatg gtgaatatcc
     3961 ctgcctaact ctattcacta tagaaagtac agcaaaaact attcttaaac ctaccaagcc
     4021 tcctactatc attatgaata attttatata ccacagccaa tttgttatgt taaaccaatt
     4081 ccacaaactt gcccatttat ctaattccaa taattcttgt tcattctttt cttgctggtt
     4141 ttgcgattct tcaattaagg agtgtattaa gcttgtgtaa ttgttaattt ctctgtccca
     4201 ctccatccag gtcgtgtgat tccaaatctg ttccagagat ttattactcc aactagcatt
     4261 ccaaggcaca gcagtggtgc aaatgagttt tccagagcaa ccccaaatcc ccaggagctg
     4321 ttgatccttt aggtatcttt ccacagccag gattcttgcc tggagctgct tgatgcccca
     4381 gactgtgagt tgcaacagat gctgttgcgc ctcaatagcc ctcagcaaat tgttctgctg
     4441 ctgcactata ccagacaata attgtctggc ctgtaccgtc agcgtcattg acgctgcgcc
     4501 catagtgctt cctgctgctc ccaagaaccc aaggaacaaa gctcctattc ccactgctct
     4561 tttttctctc tgcaccactc ttctctttgc cttggtgggt gctactccta atggttcaat
     4621 ttttactact ttatatttat ataattcact tctccaattg tccctcatat ctcctcctta
     4681 tggtgtttta ctaaactttt ccatgttcta atcctcatcc tgtctacttg ccacacaatc
     4741 atcacctgcc atctgttttc cataatccct aatgatcttt gcttttcttc ttggcactac
     4801 ttttatgtca ctattatctt gtattactac tgccccttca cctttccaga ggagctttgc
     4861 tggtcctttc caaagtggat ttctgctgtc cctgtaataa acccgaaaat tttgaatttt
     4921 tgtaatttgt ttttgtaatt ctttagtttg tatgtctgtt gctattatgt ctactattct
     4981 ttcccctgca ctgtaccccc caatcccccc ttttctttta aaattgtgga tgaatactgc
     5041 catttgtact gctgtcttaa gatgttcagc ctgatctctt acctgtccta taattttctt
     5101 taattcttta ttcatagatt ctactactcc ttgactttgg ggattgtagg gaatttccag
     5161 gtctgaagat cctgctgctt gctgtgcttt tttcttactt ttgttttgct cttcctctat
     5221 cttgtctaaa gcttccttgg tgtcttttat ctctatcctt tgatgcacac aatagagggt
     5281 tgctactgta ttatataatg atctaagttc ttctgatcct gtctgaaggg atggttgtag
     5341 ctgtcccagt atttgtctac agccttctga tgtttctaac aggccagtat taactgcgaa
     5401 tcgttctagc tccctgcttg cccatactat atgttttaat ttatattttt tctttccccc
     5461 tggccttaac cgaatttttt cccatcgcga tctaattctc ccccgcttaa tactgacgct
     5521 ctcgcaccca tctctctcct tctagcctcc gctagtcaaa atttttggcg tactcaccag
     5581 tcgccgcccc tcgcctcttg ccgtgcgcgc ttcagcaagc cgagtcctgc gtcgagagag
     5641 ctcctctggt ttccctttcg ctttcaagtc cctgttcggg cgccactgct agagattttc
     5701 cacactgact aaaagggtct gagggatctc tagttaccag agtcacacaa cagacgggca
     5761 cacactactt gaagcactca aggcaagctt tattgaggct taagcagtgg gttccctagt
     5821 tagccagaga gctcccaggc tcagatctgg tctaaccaga gagacccagt acaggcaaaa
     5881 agcagctgct tatatgcagg atctgagggc tcgccactcc ccagtcccgc ccaggccacg
     5941 cctcccatgg taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat
     6001 aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg
     6061 ggcgtacttg gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac
     6121 tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt
     6181 attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt
     6241 tatgtaacgc ggaactccat atatgggcta tgaactaatg accccgtaat tgattactat
     6301 taataactag tcaataatca atgtcgaccc aggtggcact tttcggggaa atgtgcgcgg
     6361 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     6421 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     6481 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     6541 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     6601 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     6661 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     6721 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     6781 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     6841 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     6901 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     6961 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     7021 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     7081 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     7141 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     7201 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     7261 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     7321 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     7381 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     7441 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt c
//
