LOCUS       sequence-488234-        7836 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            Vidr_004, derived from human protein CMTA2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7836)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7836)
  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_004
FEATURES             Location/Qualifiers
     source          1..7836
                     /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             2493..2504
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(join(2867..3577,3578..3580,3581..3583))
                     /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(2909..2930)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3258..3277
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3517..3538
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3580..3589
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3665..3868)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3694..3714)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3869..4172
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4429..4473)
                     /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    4658..4891
                     /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    5251..5368
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5860..5985
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6032..6289
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6291..6670
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6701..6805
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6806..6874,6875..7666)
                     /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(7024..7043)
                     /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 cacccactgg gccgccaaaa agtgcttccc
     2101 cacgtcccac acggctggaa ggctccagcg gcgctggaga cggaggtgga gatggtgggg
     2161 gagacggaat aggaggtgct gctccatgac cctttaattc ctcaccacta tcatcatctt
     2221 gaatgaaaaa gcagtttcca cggcgcccgc cagcaacgct ttggggagga ggtccacgtg
     2281 cagcggcttg cggctctaac gctgccgcag gctccaacgc actacatgga gtgtgagcgg
     2341 cttcggcttc tgggaaatcg ggactgactc cctggcctcc tccataagtc tgtccgcgct
     2401 gcggcgagtt caggaagcgg tccggatcaa aagcaggact tggcggcgcc gggggtgcac
     2461 ttggttctgt tccaacaact actgccaagc tcattgaggg gcgcggatct gcctgagggg
     2521 caaggtgacg agtaggcgtt aatccaccag cacgctgttc cagtccggtc aaaagcagaa
     2581 ttgccgttcc accgcgcgaa ctgccaccgc ggcttgtcgg gggactcgga cggatctcca
     2641 atggctccgc aaaacccgaa gaactacttg aagaactgct actcggtgaa gtgtgcgctt
     2701 tgggaagttc aggcggcagt ggagcgatca gtggaggtgg ttcggggggg tgggcgtgcg
     2761 gaatactggt caatgtcaga gcacgtggtt caactttggg tgagataatg cgatgtttgg
     2821 ttgaccggga agcggccgca cttcccccgg agcctccgct tccgcccttg tacagctcgt
     2881 ccatgccgag agtgatcccg gcggcggtca cgaactccag caggaccatg tgatcgcgct
     2941 tctcgttggg gtctttgctc agcttggact gggtgctcag gtagtggttg tcgggcagca
     3001 gcacggggcc gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc tgcacgctgc
     3061 cgtcctcgat gttgtggcgg atcttgaagt tcaccttgat gccgttcttc tgcttgtcgg
     3121 ccatgatata gacgttgtgg ctgttgtagt tgtactccag cttgtgcccc aggatgttgc
     3181 cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt caccagggtg tcgccctcga
     3241 acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct
     3301 ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg tggtcggggt
     3361 agcggctgaa gcactgcacg ccgtaggtca gggtggtcac gagggtgggc cagggcacgg
     3421 gcagcttgcc ggtggtgcag atgaacttca gggtcagctt gccgtaggtg gcatcgccct
     3481 cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc gccgtccagc tcgaccagga
     3541 tgggcaccac cccggtgaac agctcctcgc ccttgctcac catggtggcg accggtggat
     3601 cccgggcccg cggtaccgtc gactgcagaa ttcggttcac taaacgagcc cggttcacta
     3661 aacgagctct gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg
     3721 gggcggagtt gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc
     3781 cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc
     3841 cattgatgta ctgccaaaac cgcatcacca tggtaatagc gatgactaat acgtagatgt
     3901 actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt
     3961 taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca
     4021 agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg
     4081 ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc
     4141 caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta
     4201 atgaccccgt aattgattac tattaataac tagcacgcgt tcgaagcttg agctcggtcc
     4261 cctcggggtt gggaggtggg tctgaaacga taatggtgaa tatccctgcc taactctatt
     4321 cactatagaa agtacagcaa aaactattct taaacctacc aagcctccta ctatcattat
     4381 gaataatttt atataccaca gccaatttgt tatgttaaac caattccaca aacttgccca
     4441 tttatctaat tccaataatt cttgttcatt cttttcttgc tggttttgcg attcttcaat
     4501 taaggagtgt attaagcttg tgtaattgtt aatttctctg tcccactcca tccaggtcgt
     4561 gtgattccaa atctgttcca gagatttatt actccaacta gcattccaag gcacagcagt
     4621 ggtgcaaatg agttttccag agcaacccca aatccccagg agctgttgat cctttaggta
     4681 tctttccaca gccaggattc ttgcctggag ctgcttgatg ccccagactg tgagttgcaa
     4741 cagatgctgt tgcgcctcaa tagccctcag caaattgttc tgctgctgca ctataccaga
     4801 caataattgt ctggcctgta ccgtcagcgt cattgacgct gcgcccatag tgcttcctgc
     4861 tgctcccaag aacccaagga acaaagctcc tattcccact gctctttttt ctctctgcac
     4921 cactcttctc tttgccttgg tgggtgctac tcctaatggt tcaattttta ctactttata
     4981 tttatataat tcacttctcc aattgtccct catatctcct ccttatggtg ttttactaaa
     5041 cttttccatg ttctaatcct catcctgtct acttgccaca caatcatcac ctgccatctg
     5101 ttttccataa tccctaatga tctttgcttt tcttcttggc actactttta tgtcactatt
     5161 atcttgtatt actactgccc cttcaccttt ccagaggagc tttgctggtc ctttccaaag
     5221 tggatttctg ctgtccctgt aataaacccg aaaattttga atttttgtaa tttgtttttg
     5281 taattcttta gtttgtatgt ctgttgctat tatgtctact attctttccc ctgcactgta
     5341 ccccccaatc cccccttttc ttttaaaatt gtggatgaat actgccattt gtactgctgt
     5401 cttaagatgt tcagcctgat ctcttacctg tcctataatt ttctttaatt ctttattcat
     5461 agattctact actccttgac tttggggatt gtagggaatt tccaggtctg aagatcctgc
     5521 tgcttgctgt gcttttttct tacttttgtt ttgctcttcc tctatcttgt ctaaagcttc
     5581 cttggtgtct tttatctcta tcctttgatg cacacaatag agggttgcta ctgtattata
     5641 taatgatcta agttcttctg atcctgtctg aagggatggt tgtagctgtc ccagtatttg
     5701 tctacagcct tctgatgttt ctaacaggcc agtattaact gcgaatcgtt ctagctccct
     5761 gcttgcccat actatatgtt ttaatttata ttttttcttt ccccctggcc ttaaccgaat
     5821 tttttcccat cgcgatctaa ttctcccccg cttaatactg acgctctcgc acccatctct
     5881 ctccttctag cctccgctag tcaaaatttt tggcgtactc accagtcgcc gcccctcgcc
     5941 tcttgccgtg cgcgcttcag caagccgagt cctgcgtcga gagagctcct ctggtttccc
     6001 tttcgctttc aagtccctgt tcgggcgcca ctgctagaga ttttccacac tgactaaaag
     6061 ggtctgaggg atctctagtt accagagtca cacaacagac gggcacacac tacttgaagc
     6121 actcaaggca agctttattg aggcttaagc agtgggttcc ctagttagcc agagagctcc
     6181 caggctcaga tctggtctaa ccagagagac ccagtacagg caaaaagcag ctgcttatat
     6241 gcaggatctg agggctcgcc actccccagt cccgcccagg ccacgcctcc catggtaata
     6301 gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg
     6361 gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata
     6421 tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt
     6481 caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggg
     6541 cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgcggaac
     6601 tccatatatg ggctatgaac taatgacccc gtaattgatt actattaata actagtcaat
     6661 aatcaatgtc gacccaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     6721 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     6781 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     6841 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     6901 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     6961 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     7021 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     7081 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     7141 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     7201 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     7261 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     7321 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     7381 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     7441 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     7501 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     7561 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     7621 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     7681 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     7741 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     7801 agcgtcagac cccgtagaaa agatcaaagg atcttc
//
