LOCUS       sequence-488258-        8370 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            Vidr_029, derived from human protein DLGP3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8370)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8370)
  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_029
FEATURES             Location/Qualifiers
     source          1..8370
                     /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(2657..2686)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             complement(2660..2689)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             complement(2663..2692)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             complement(join(3401..4111,4112..4114,4115..4117))
                     /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(3443..3464)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3792..3811
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4051..4072
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4114..4123
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4199..4402)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4228..4248)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4403..4706
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4963..5007)
                     /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    5192..5425
                     /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    5785..5902
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6394..6519
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6566..6823
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6825..7204
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7235..7339
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7340..7408,7409..8200)
                     /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(7558..7577)
                     /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 ccggttcaag agctacgacg cgtggtaaag cggcgggcca
     2101 cagccttggg gctagtcttg gggcttccat cactgtcacc cgactcttca tcccccattg
     2161 ctttaatgta gctaccagag cgcatacgac gacaggggat ttcaccatcc ttgccacctg
     2221 tggggtaacc gccccagtca tcctggggca cctgaaggta gtggtacgtg cgtgctttcg
     2281 cctttgtctc cgggcctaat aaccctttcc ccggaccagc tccaggatac ccgtcacgtc
     2341 cttggctaac catcattgta tgccacgcag agcgcttaac gctttgaccg tccagggaca
     2401 tcgacatgcc ggtacaagca aggcaacgcc cttccgaacc tcccgagcgg cccttaaagc
     2461 tcaagtcgcg gtaggagcca tctggccctt caaggcaaaa gggaccgcca ggttctccgg
     2521 gcggacgccc accggccagg aatccggaat cagagtccag attgtcgtct gaagaccacc
     2581 acccggtgga cttcgcttga tggcgaccat cccccttgcg atccttactt ttactgcgtt
     2641 ttccatgacg ggactgatgg tgatggtggt gatgatgatg gtgatgatga tggcttgtgt
     2701 gtggtccccc actaccgggc ccgggatagg aatctccccc actaccgccg cgaccctccg
     2761 ctttgggccc gttgtaatca cgtttgccag gggcttccag cgagtgcgat ttagcaaaca
     2821 gtttctgaac agagtgtact aagtggcgaa tgcgagatgg gctctcacta cggggttctg
     2881 gagcggtccc tgtccctggt gcaggacctg gaccggcgcc ggcagggcca cgttggtacg
     2941 gcaaagtatg gaagccatcc tgttgaacag gaagttgttt ttcaaattga tcaagcaaag
     3001 tcggcggtaa acgcggggcg cccttcccct gggggtgtcc gacacaatcc tcgcatgtat
     3061 caaatggacc ttgacctggg tacatgcgcg ggaacgttga actcccaccc ccaacgccag
     3121 ctcctgcagg ccccccttcc gggccaacac ttggcccttc ggacagggac aaaggccctt
     3181 caggactaat atggcccaag cctgcacgcg gagcacaaaa gcgaggttca gtggagaagg
     3241 cctcgcgaga acccagaagg tatggcgcac gtgcagcggg acccacgtcc atgtgttgtt
     3301 ggtccgcaaa gcgagcggga cgcgggtgcg atccgcgatc cccgtgataa ccgcgcatcc
     3361 gggaagcggc cgcacttccc ccggagcctc cgcttccgcc cttgtacagc tcgtccatgc
     3421 cgagagtgat cccggcggcg gtcacgaact ccagcaggac catgtgatcg cgcttctcgt
     3481 tggggtcttt gctcagcttg gactgggtgc tcaggtagtg gttgtcgggc agcagcacgg
     3541 ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc gagctgcacg ctgccgtcct
     3601 cgatgttgtg gcggatcttg aagttcacct tgatgccgtt cttctgcttg tcggccatga
     3661 tatagacgtt gtggctgttg tagttgtact ccagcttgtg ccccaggatg ttgccgtcct
     3721 ccttgaagtc gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca
     3781 cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt
     3841 agccttcggg catggcggac ttgaagaagt cgtgctgctt catgtggtcg gggtagcggc
     3901 tgaagcactg cacgccgtag gtcagggtgg tcacgagggt gggccagggc acgggcagct
     3961 tgccggtggt gcagatgaac ttcagggtca gcttgccgta ggtggcatcg ccctcgccct
     4021 cgccggacac gctgaacttg tggccgttta cgtcgccgtc cagctcgacc aggatgggca
     4081 ccaccccggt gaacagctcc tcgcccttgc tcaccatggt ggcgaccggt ggatcccggg
     4141 cccgcggtac cgtcgactgc agaattcggt tcactaaacg agcccggttc actaaacgag
     4201 ctctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg
     4261 agttgttacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga
     4321 cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga
     4381 tgtactgcca aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc
     4441 aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt
     4501 cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg
     4561 cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta
     4621 tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg
     4681 ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc
     4741 ccgtaattga ttactattaa taactagcac gcgttcgaag cttgagctcg gtcccctcgg
     4801 ggttgggagg tgggtctgaa acgataatgg tgaatatccc tgcctaactc tattcactat
     4861 agaaagtaca gcaaaaacta ttcttaaacc taccaagcct cctactatca ttatgaataa
     4921 ttttatatac cacagccaat ttgttatgtt aaaccaattc cacaaacttg cccatttatc
     4981 taattccaat aattcttgtt cattcttttc ttgctggttt tgcgattctt caattaagga
     5041 gtgtattaag cttgtgtaat tgttaatttc tctgtcccac tccatccagg tcgtgtgatt
     5101 ccaaatctgt tccagagatt tattactcca actagcattc caaggcacag cagtggtgca
     5161 aatgagtttt ccagagcaac cccaaatccc caggagctgt tgatccttta ggtatctttc
     5221 cacagccagg attcttgcct ggagctgctt gatgccccag actgtgagtt gcaacagatg
     5281 ctgttgcgcc tcaatagccc tcagcaaatt gttctgctgc tgcactatac cagacaataa
     5341 ttgtctggcc tgtaccgtca gcgtcattga cgctgcgccc atagtgcttc ctgctgctcc
     5401 caagaaccca aggaacaaag ctcctattcc cactgctctt ttttctctct gcaccactct
     5461 tctctttgcc ttggtgggtg ctactcctaa tggttcaatt tttactactt tatatttata
     5521 taattcactt ctccaattgt ccctcatatc tcctccttat ggtgttttac taaacttttc
     5581 catgttctaa tcctcatcct gtctacttgc cacacaatca tcacctgcca tctgttttcc
     5641 ataatcccta atgatctttg cttttcttct tggcactact tttatgtcac tattatcttg
     5701 tattactact gccccttcac ctttccagag gagctttgct ggtcctttcc aaagtggatt
     5761 tctgctgtcc ctgtaataaa cccgaaaatt ttgaattttt gtaatttgtt tttgtaattc
     5821 tttagtttgt atgtctgttg ctattatgtc tactattctt tcccctgcac tgtacccccc
     5881 aatcccccct tttcttttaa aattgtggat gaatactgcc atttgtactg ctgtcttaag
     5941 atgttcagcc tgatctctta cctgtcctat aattttcttt aattctttat tcatagattc
     6001 tactactcct tgactttggg gattgtaggg aatttccagg tctgaagatc ctgctgcttg
     6061 ctgtgctttt ttcttacttt tgttttgctc ttcctctatc ttgtctaaag cttccttggt
     6121 gtcttttatc tctatccttt gatgcacaca atagagggtt gctactgtat tatataatga
     6181 tctaagttct tctgatcctg tctgaaggga tggttgtagc tgtcccagta tttgtctaca
     6241 gccttctgat gtttctaaca ggccagtatt aactgcgaat cgttctagct ccctgcttgc
     6301 ccatactata tgttttaatt tatatttttt ctttccccct ggccttaacc gaattttttc
     6361 ccatcgcgat ctaattctcc cccgcttaat actgacgctc tcgcacccat ctctctcctt
     6421 ctagcctccg ctagtcaaaa tttttggcgt actcaccagt cgccgcccct cgcctcttgc
     6481 cgtgcgcgct tcagcaagcc gagtcctgcg tcgagagagc tcctctggtt tccctttcgc
     6541 tttcaagtcc ctgttcgggc gccactgcta gagattttcc acactgacta aaagggtctg
     6601 agggatctct agttaccaga gtcacacaac agacgggcac acactacttg aagcactcaa
     6661 ggcaagcttt attgaggctt aagcagtggg ttccctagtt agccagagag ctcccaggct
     6721 cagatctggt ctaaccagag agacccagta caggcaaaaa gcagctgctt atatgcagga
     6781 tctgagggct cgccactccc cagtcccgcc caggccacgc ctcccatggt aatagcgatg
     6841 actaatacgt agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa
     6901 tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac
     6961 acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg
     7021 aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg
     7081 tcgttgggcg gtcagccagg cgggccattt accgtaagtt atgtaacgcg gaactccata
     7141 tatgggctat gaactaatga ccccgtaatt gattactatt aataactagt caataatcaa
     7201 tgtcgaccca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     7261 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     7321 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     7381 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     7441 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     7501 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     7561 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     7621 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     7681 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     7741 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     7801 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     7861 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     7921 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     7981 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     8041 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     8101 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     8161 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     8221 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     8281 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     8341 agaccccgta gaaaagatca aaggatcttc
//
