LOCUS       sequence-488233-        7758 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            Vidr_003, derived from human protein EFCB5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7758)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7758)
  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_003
FEATURES             Location/Qualifiers
     source          1..7758
                     /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(2789..3499,3500..3502,3503..3505))
                     /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(2831..2852)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3180..3199
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3439..3460
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3502..3511
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3587..3790)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3616..3636)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3791..4094
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4351..4395)
                     /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    4580..4813
                     /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    5173..5290
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5782..5907
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5954..6211
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6213..6592
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6623..6727
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6728..6796,6797..7588)
                     /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(6946..6965)
                     /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 ccggttcatt cttgtaaggg cacttcgata tattctgatg
     2101 tgatgggttc tccatactta gtgctcttca agatagaatc tttacgggat tgactagtag
     2161 atccgatgtc ttcttgttct tcagagatga tttccccgta tggcccttgc tcgctcttct
     2221 ggtacggttc ttcaataaca gactcacgaa gtgagccctg ctcggctgca gacatacggc
     2281 gcgaaccctg ttccgcaacg ctccctttgt gacgatcttg ctcagcgatt gactcacgac
     2341 gagaaccctg ctcagcaacc gactcgcggc tacttccctg ctccgagacg ctgacgcggc
     2401 ggggtccctg gccctcaata ctacccttgt gttggccctg ctcggtcgtg ctttcacgat
     2461 gcgtttcctg ttctgttaac aagcggcgac tggagccttg ctctaatgta gattcggtgt
     2521 gtgttccggg ctcgataacg ctctcaatat acagctcttg ttcagccgta ggcttcttcc
     2581 cctgctgttg ctcctcaata ctaatacgct ggggaccctg ctcggcggtg accccgcgct
     2641 gctgctcggg ggggtttgga cttggtgtgc tggtacgttg ttcgtttaac ttaggttggt
     2701 cgggtgattc aagtaattta ctttgagtct ttgaggcatg gttcgcccgg gaagcggccg
     2761 cacttccccc ggagcctccg cttccgccct tgtacagctc gtccatgccg agagtgatcc
     2821 cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc
     2881 tcagcttgga ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga
     2941 tgggggtgtt ctgctggtag tggtcggcga gctgcacgct gccgtcctcg atgttgtggc
     3001 ggatcttgaa gttcaccttg atgccgttct tctgcttgtc ggccatgata tagacgttgt
     3061 ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga
     3121 tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg
     3181 tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca
     3241 tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg aagcactgca
     3301 cgccgtaggt cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc
     3361 agatgaactt cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc
     3421 tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga
     3481 acagctcctc gcccttgctc accatggtgg cgaccggtgg atcccgggcc cgcggtaccg
     3541 tcgactgcag aattcggttc actaaacgag cccggttcac taaacgagct ctgcttatat
     3601 agacctccca ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag ttgttacgac
     3661 attttggaaa gtcccgttga ttttggtgcc aaaacaaact cccattgacg tcaatggggt
     3721 ggagacttgg aaatccccgt gagtcaaacc gctatccacg cccattgatg tactgccaaa
     3781 accgcatcac catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt
     3841 cccataaggt catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa
     3901 tagggggcgt acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta
     3961 aatactccac ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg
     4021 tcattattga cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg
     4081 taagttatgt aacgcggaac tccatatatg ggctatgaac taatgacccc gtaattgatt
     4141 actattaata actagcacgc gttcgaagct tgagctcggt cccctcgggg ttgggaggtg
     4201 ggtctgaaac gataatggtg aatatccctg cctaactcta ttcactatag aaagtacagc
     4261 aaaaactatt cttaaaccta ccaagcctcc tactatcatt atgaataatt ttatatacca
     4321 cagccaattt gttatgttaa accaattcca caaacttgcc catttatcta attccaataa
     4381 ttcttgttca ttcttttctt gctggttttg cgattcttca attaaggagt gtattaagct
     4441 tgtgtaattg ttaatttctc tgtcccactc catccaggtc gtgtgattcc aaatctgttc
     4501 cagagattta ttactccaac tagcattcca aggcacagca gtggtgcaaa tgagttttcc
     4561 agagcaaccc caaatcccca ggagctgttg atcctttagg tatctttcca cagccaggat
     4621 tcttgcctgg agctgcttga tgccccagac tgtgagttgc aacagatgct gttgcgcctc
     4681 aatagccctc agcaaattgt tctgctgctg cactatacca gacaataatt gtctggcctg
     4741 taccgtcagc gtcattgacg ctgcgcccat agtgcttcct gctgctccca agaacccaag
     4801 gaacaaagct cctattccca ctgctctttt ttctctctgc accactcttc tctttgcctt
     4861 ggtgggtgct actcctaatg gttcaatttt tactacttta tatttatata attcacttct
     4921 ccaattgtcc ctcatatctc ctccttatgg tgttttacta aacttttcca tgttctaatc
     4981 ctcatcctgt ctacttgcca cacaatcatc acctgccatc tgttttccat aatccctaat
     5041 gatctttgct tttcttcttg gcactacttt tatgtcacta ttatcttgta ttactactgc
     5101 cccttcacct ttccagagga gctttgctgg tcctttccaa agtggatttc tgctgtccct
     5161 gtaataaacc cgaaaatttt gaatttttgt aatttgtttt tgtaattctt tagtttgtat
     5221 gtctgttgct attatgtcta ctattctttc ccctgcactg taccccccaa tccccccttt
     5281 tcttttaaaa ttgtggatga atactgccat ttgtactgct gtcttaagat gttcagcctg
     5341 atctcttacc tgtcctataa ttttctttaa ttctttattc atagattcta ctactccttg
     5401 actttgggga ttgtagggaa tttccaggtc tgaagatcct gctgcttgct gtgctttttt
     5461 cttacttttg ttttgctctt cctctatctt gtctaaagct tccttggtgt cttttatctc
     5521 tatcctttga tgcacacaat agagggttgc tactgtatta tataatgatc taagttcttc
     5581 tgatcctgtc tgaagggatg gttgtagctg tcccagtatt tgtctacagc cttctgatgt
     5641 ttctaacagg ccagtattaa ctgcgaatcg ttctagctcc ctgcttgccc atactatatg
     5701 ttttaattta tattttttct ttccccctgg ccttaaccga attttttccc atcgcgatct
     5761 aattctcccc cgcttaatac tgacgctctc gcacccatct ctctccttct agcctccgct
     5821 agtcaaaatt tttggcgtac tcaccagtcg ccgcccctcg cctcttgccg tgcgcgcttc
     5881 agcaagccga gtcctgcgtc gagagagctc ctctggtttc cctttcgctt tcaagtccct
     5941 gttcgggcgc cactgctaga gattttccac actgactaaa agggtctgag ggatctctag
     6001 ttaccagagt cacacaacag acgggcacac actacttgaa gcactcaagg caagctttat
     6061 tgaggcttaa gcagtgggtt ccctagttag ccagagagct cccaggctca gatctggtct
     6121 aaccagagag acccagtaca ggcaaaaagc agctgcttat atgcaggatc tgagggctcg
     6181 ccactcccca gtcccgccca ggccacgcct cccatggtaa tagcgatgac taatacgtag
     6241 atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc
     6301 catttaccgt cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact
     6361 gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt
     6421 ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt
     6481 cagccaggcg ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga
     6541 actaatgacc ccgtaattga ttactattaa taactagtca ataatcaatg tcgacccagg
     6601 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     6661 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     6721 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     6781 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     6841 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     6901 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     6961 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     7021 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     7081 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     7141 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     7201 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     7261 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     7321 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     7381 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     7441 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     7501 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     7561 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     7621 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     7681 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     7741 aaagatcaaa ggatcttc
//
