LOCUS       sequence-488238-        7305 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            Vidr_008, derived from human protein GCM2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7305)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7305)
  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_008
FEATURES             Location/Qualifiers
     source          1..7305
                     /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(2336..3046,3047..3049,3050..3052))
                     /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(2378..2399)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2727..2746
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2986..3007
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3049..3058
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3134..3337)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3163..3183)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3338..3641
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3898..3942)
                     /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    4127..4360
                     /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    4720..4837
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5329..5454
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5501..5758
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5760..6139
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6170..6274
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6275..6343,6344..7135)
                     /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(6493..6512)
                     /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 ccggttcatg ggatgggaaa ggaggtctcg gtaacgatat
     2101 cgaaatcttc cgggttttcc aaggggggaa tattggaaaa atgtccactg gagtcttgat
     2161 tttcttcggc ctcggattca cggatacgct ttttctgggg ctggtagaag gaagccatct
     2221 ggcgcttgat agctgagcgg cgtgcctctg tctctgactt tgattccggg cgaggatggt
     2281 cgtgtacccc cttccgggaa gcggccgcac ttcccccgga gcctccgctt ccgcccttgt
     2341 acagctcgtc catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt
     2401 gatcgcgctt ctcgttgggg tctttgctca gcttggactg ggtgctcagg tagtggttgt
     2461 cgggcagcag cacggggccg tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct
     2521 gcacgctgcc gtcctcgatg ttgtggcgga tcttgaagtt caccttgatg ccgttcttct
     2581 gcttgtcggc catgatatag acgttgtggc tgttgtagtt gtactccagc ttgtgcccca
     2641 ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc accagggtgt
     2701 cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg
     2761 tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt
     2821 ggtcggggta gcggctgaag cactgcacgc cgtaggtcag ggtggtcacg agggtgggcc
     2881 agggcacggg cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg
     2941 catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg ccgtccagct
     3001 cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc atggtggcga
     3061 ccggtggatc ccgggcccgc ggtaccgtcg actgcagaat tcggttcact aaacgagccc
     3121 ggttcactaa acgagctctg cttatataga cctcccaccg tacacgccta ccgcccattt
     3181 gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa
     3241 acaaactccc attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct
     3301 atccacgccc attgatgtac tgccaaaacc gcatcaccat ggtaatagcg atgactaata
     3361 cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg
     3421 cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat
     3481 gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc
     3541 ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg
     3601 gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc
     3661 tatgaactaa tgaccccgta attgattact attaataact agcacgcgtt cgaagcttga
     3721 gctcggtccc ctcggggttg ggaggtgggt ctgaaacgat aatggtgaat atccctgcct
     3781 aactctattc actatagaaa gtacagcaaa aactattctt aaacctacca agcctcctac
     3841 tatcattatg aataatttta tataccacag ccaatttgtt atgttaaacc aattccacaa
     3901 acttgcccat ttatctaatt ccaataattc ttgttcattc ttttcttgct ggttttgcga
     3961 ttcttcaatt aaggagtgta ttaagcttgt gtaattgtta atttctctgt cccactccat
     4021 ccaggtcgtg tgattccaaa tctgttccag agatttatta ctccaactag cattccaagg
     4081 cacagcagtg gtgcaaatga gttttccaga gcaaccccaa atccccagga gctgttgatc
     4141 ctttaggtat ctttccacag ccaggattct tgcctggagc tgcttgatgc cccagactgt
     4201 gagttgcaac agatgctgtt gcgcctcaat agccctcagc aaattgttct gctgctgcac
     4261 tataccagac aataattgtc tggcctgtac cgtcagcgtc attgacgctg cgcccatagt
     4321 gcttcctgct gctcccaaga acccaaggaa caaagctcct attcccactg ctcttttttc
     4381 tctctgcacc actcttctct ttgccttggt gggtgctact cctaatggtt caatttttac
     4441 tactttatat ttatataatt cacttctcca attgtccctc atatctcctc cttatggtgt
     4501 tttactaaac ttttccatgt tctaatcctc atcctgtcta cttgccacac aatcatcacc
     4561 tgccatctgt tttccataat ccctaatgat ctttgctttt cttcttggca ctacttttat
     4621 gtcactatta tcttgtatta ctactgcccc ttcacctttc cagaggagct ttgctggtcc
     4681 tttccaaagt ggatttctgc tgtccctgta ataaacccga aaattttgaa tttttgtaat
     4741 ttgtttttgt aattctttag tttgtatgtc tgttgctatt atgtctacta ttctttcccc
     4801 tgcactgtac cccccaatcc ccccttttct tttaaaattg tggatgaata ctgccatttg
     4861 tactgctgtc ttaagatgtt cagcctgatc tcttacctgt cctataattt tctttaattc
     4921 tttattcata gattctacta ctccttgact ttggggattg tagggaattt ccaggtctga
     4981 agatcctgct gcttgctgtg cttttttctt acttttgttt tgctcttcct ctatcttgtc
     5041 taaagcttcc ttggtgtctt ttatctctat cctttgatgc acacaataga gggttgctac
     5101 tgtattatat aatgatctaa gttcttctga tcctgtctga agggatggtt gtagctgtcc
     5161 cagtatttgt ctacagcctt ctgatgtttc taacaggcca gtattaactg cgaatcgttc
     5221 tagctccctg cttgcccata ctatatgttt taatttatat tttttctttc cccctggcct
     5281 taaccgaatt ttttcccatc gcgatctaat tctcccccgc ttaatactga cgctctcgca
     5341 cccatctctc tccttctagc ctccgctagt caaaattttt ggcgtactca ccagtcgccg
     5401 cccctcgcct cttgccgtgc gcgcttcagc aagccgagtc ctgcgtcgag agagctcctc
     5461 tggtttccct ttcgctttca agtccctgtt cgggcgccac tgctagagat tttccacact
     5521 gactaaaagg gtctgaggga tctctagtta ccagagtcac acaacagacg ggcacacact
     5581 acttgaagca ctcaaggcaa gctttattga ggcttaagca gtgggttccc tagttagcca
     5641 gagagctccc aggctcagat ctggtctaac cagagagacc cagtacaggc aaaaagcagc
     5701 tgcttatatg caggatctga gggctcgcca ctccccagtc ccgcccaggc cacgcctccc
     5761 atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc
     5821 atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta
     5881 cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc
     5941 cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac
     6001 gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta
     6061 acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa
     6121 ctagtcaata atcaatgtcg acccaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     6181 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     6241 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     6301 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     6361 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     6421 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     6481 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     6541 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     6601 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     6661 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     6721 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     6781 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     6841 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     6901 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     6961 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     7021 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     7081 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     7141 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     7201 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     7261 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttc
//
