LOCUS       sequence-488250-        7806 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            Vidr_021, derived from human protein CEP95.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7806)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7806)
  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_021
FEATURES             Location/Qualifiers
     source          1..7806
                     /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(2837..3547,3548..3550,3551..3553))
                     /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(2879..2900)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3228..3247
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3487..3508
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3550..3559
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3635..3838)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3664..3684)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3839..4142
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4399..4443)
                     /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    4628..4861
                     /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    5221..5338
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5830..5955
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6002..6259
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6261..6640
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6671..6775
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6776..6844,6845..7636)
                     /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(6994..7013)
                     /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 ccggttcaac taactttcat tggaactgcc ttatttggtt
     2101 tgggcaatcc cccacgcaga ctctgtgttg tggttttacg actataaatc ttccacggct
     2161 gactgcattc gggagtaccc ttacggactg cttctccacg gtagatcttc tcagtttctt
     2221 cttctttctc gtggatgcga agcggaccaa tgttctcctg aaccttgtgg cggcgtaatt
     2281 cgcgctcgaa agcctcagta aatgcttgtg cctgaaattg acgtacatct gtctcgcgcg
     2341 gtttgcgctg gcgcttacgt tccagatgga actgcttatg cttatttact ggggatgggg
     2401 acaggctatg ggagcgatag gggggcttgc ggcgcatcga caacccagga cgtgactttt
     2461 ttggtccgta ctcgacaata ccgtctgaat gctggctcaa agtctcttca gtgccgtcct
     2521 ccacctcctc gtttcctgtg ttctcctctt tatgatcggt tttctccttg atacgatcgc
     2581 ccaacgcaga tttcagcatc cagtccagct cagaaaggcg ttgcgacagt ttttcggaga
     2641 catcatgaag ttcttgctct gtcaggcgct tgcgaggacg ctgaggaaac ggggagttgc
     2701 atgaggatgc agttgcacga ttctcgttac gttttccttt gggcggctta cgtgtgcgag
     2761 ggccctgaac ttgggcaggg tacacctccc acttccggga agcggccgca cttcccccgg
     2821 agcctccgct tccgcccttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca
     2881 cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc agcttggact
     2941 gggtgctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct
     3001 gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt
     3061 tcaccttgat gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt
     3121 tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct
     3181 cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc
     3241 cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga
     3301 agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg ccgtaggtca
     3361 gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atgaacttca
     3421 gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc
     3481 cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc
     3541 ccttgctcac catggtggcg accggtggat cccgggcccg cggtaccgtc gactgcagaa
     3601 ttcggttcac taaacgagcc cggttcacta aacgagctct gcttatatag acctcccacc
     3661 gtacacgcct accgcccatt tgcgtcaatg gggcggagtt gttacgacat tttggaaagt
     3721 cccgttgatt ttggtgccaa aacaaactcc cattgacgtc aatggggtgg agacttggaa
     3781 atccccgtga gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac cgcatcacca
     3841 tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca
     3901 tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac
     3961 ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc
     4021 attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg
     4081 tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa
     4141 cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac tattaataac
     4201 tagcacgcgt tcgaagcttg agctcggtcc cctcggggtt gggaggtggg tctgaaacga
     4261 taatggtgaa tatccctgcc taactctatt cactatagaa agtacagcaa aaactattct
     4321 taaacctacc aagcctccta ctatcattat gaataatttt atataccaca gccaatttgt
     4381 tatgttaaac caattccaca aacttgccca tttatctaat tccaataatt cttgttcatt
     4441 cttttcttgc tggttttgcg attcttcaat taaggagtgt attaagcttg tgtaattgtt
     4501 aatttctctg tcccactcca tccaggtcgt gtgattccaa atctgttcca gagatttatt
     4561 actccaacta gcattccaag gcacagcagt ggtgcaaatg agttttccag agcaacccca
     4621 aatccccagg agctgttgat cctttaggta tctttccaca gccaggattc ttgcctggag
     4681 ctgcttgatg ccccagactg tgagttgcaa cagatgctgt tgcgcctcaa tagccctcag
     4741 caaattgttc tgctgctgca ctataccaga caataattgt ctggcctgta ccgtcagcgt
     4801 cattgacgct gcgcccatag tgcttcctgc tgctcccaag aacccaagga acaaagctcc
     4861 tattcccact gctctttttt ctctctgcac cactcttctc tttgccttgg tgggtgctac
     4921 tcctaatggt tcaattttta ctactttata tttatataat tcacttctcc aattgtccct
     4981 catatctcct ccttatggtg ttttactaaa cttttccatg ttctaatcct catcctgtct
     5041 acttgccaca caatcatcac ctgccatctg ttttccataa tccctaatga tctttgcttt
     5101 tcttcttggc actactttta tgtcactatt atcttgtatt actactgccc cttcaccttt
     5161 ccagaggagc tttgctggtc ctttccaaag tggatttctg ctgtccctgt aataaacccg
     5221 aaaattttga atttttgtaa tttgtttttg taattcttta gtttgtatgt ctgttgctat
     5281 tatgtctact attctttccc ctgcactgta ccccccaatc cccccttttc ttttaaaatt
     5341 gtggatgaat actgccattt gtactgctgt cttaagatgt tcagcctgat ctcttacctg
     5401 tcctataatt ttctttaatt ctttattcat agattctact actccttgac tttggggatt
     5461 gtagggaatt tccaggtctg aagatcctgc tgcttgctgt gcttttttct tacttttgtt
     5521 ttgctcttcc tctatcttgt ctaaagcttc cttggtgtct tttatctcta tcctttgatg
     5581 cacacaatag agggttgcta ctgtattata taatgatcta agttcttctg atcctgtctg
     5641 aagggatggt tgtagctgtc ccagtatttg tctacagcct tctgatgttt ctaacaggcc
     5701 agtattaact gcgaatcgtt ctagctccct gcttgcccat actatatgtt ttaatttata
     5761 ttttttcttt ccccctggcc ttaaccgaat tttttcccat cgcgatctaa ttctcccccg
     5821 cttaatactg acgctctcgc acccatctct ctccttctag cctccgctag tcaaaatttt
     5881 tggcgtactc accagtcgcc gcccctcgcc tcttgccgtg cgcgcttcag caagccgagt
     5941 cctgcgtcga gagagctcct ctggtttccc tttcgctttc aagtccctgt tcgggcgcca
     6001 ctgctagaga ttttccacac tgactaaaag ggtctgaggg atctctagtt accagagtca
     6061 cacaacagac gggcacacac tacttgaagc actcaaggca agctttattg aggcttaagc
     6121 agtgggttcc ctagttagcc agagagctcc caggctcaga tctggtctaa ccagagagac
     6181 ccagtacagg caaaaagcag ctgcttatat gcaggatctg agggctcgcc actccccagt
     6241 cccgcccagg ccacgcctcc catggtaata gcgatgacta atacgtagat gtactgccaa
     6301 gtaggaaagt cccataaggt catgtactgg gcataatgcc aggcgggcca tttaccgtca
     6361 ttgacgtcaa tagggggcgt acttggcata tgatacactt gatgtactgc caagtgggca
     6421 gtttaccgta aatactccac ccattgacgt caatggaaag tccctattgg cgttactatg
     6481 ggaacatacg tcattattga cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg
     6541 ccatttaccg taagttatgt aacgcggaac tccatatatg ggctatgaac taatgacccc
     6601 gtaattgatt actattaata actagtcaat aatcaatgtc gacccaggtg gcacttttcg
     6661 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     6721 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     6781 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     6841 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     6901 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     6961 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     7021 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     7081 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     7141 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     7201 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     7261 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     7321 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     7381 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     7441 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     7501 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     7561 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     7621 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     7681 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     7741 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     7801 atcttc
//
