LOCUS       sequence-488257-        8859 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            Vidr_028, derived from human protein R3HD2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8859)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8859)
  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_028
FEATURES             Location/Qualifiers
     source          1..8859
                     /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(3890..4600,4601..4603,4604..4606))
                     /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(3932..3953)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4281..4300
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4540..4561
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4603..4612
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4688..4891)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4717..4737)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4892..5195
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(5452..5496)
                     /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    5681..5914
                     /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    6274..6391
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6883..7008
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             7055..7312
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        7314..7693
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7724..7828
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7829..7897,7898..8689)
                     /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(8047..8066)
                     /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 ccggttcaaa gcaatgagta gcgcccatct ccttgcgcag
     2101 gtcctccagg gcgtggaaat tgggtcagca ctggaagggg gcttaaacct gtgcaaactg
     2161 aggaaagact agtgacggga gaaggtgcag gtgactgcgt tggagaggtc accggggagg
     2221 aagacatttg ggtattggcc tgaggagatg agtcggggga atacaggtct ccaggttttt
     2281 gcccgcgctg gtccatgcta taatatttgc agtgagacca ctgtaccata ctcggatttt
     2341 gtggtggttg gggaccatta gggacattta actgcattac caccactccc ggacccgatg
     2401 gcgacacgcc agaatactgt tgaggcattt gggggggcga gcagggggac ggactctgtg
     2461 gcatttgata ttgctccgat cctggaggct gaagaaaccc cacggaagga cttgttccat
     2521 tctgttgggc cgggggaatc atactgtagt agaccggtac gcctgccgct ggcaatccgc
     2581 cttgcaccga ctggctgacg ggaaccaaca ttggttgttg aaagggaggt tgtaccacat
     2641 tctgactgtc agaacctacc ggtacttggt agctcggcaa cggggtatac tgcaccacaa
     2701 gtccctgcat ttgaccgccc atggagctgc gttgagaaga taacagcccc tgattctgcg
     2761 gttgttgtac cccaatcatc ccttgataag ctgcttgttg ttgatttggc atcatcggtt
     2821 gcagtcctgg ctgttgtgag ggttggggta actgttgtcc acgttgtggg ctgtaagcga
     2881 ccggatgact caatggacga tattgttggt tgctattagg atattgacca gggggataat
     2941 atgatacttg gatctgctgc ggtggctgca tgtatccctg tggcggaagt acttgctggg
     3001 tgggaggtgc gtgggaagat gtggaatagt tacttgtcgg cagcggctgg cccgttgaag
     3061 ccatgatgaa actggtctgc tgaggatgtt gtgagatcaa tggcgtctga aacagcgcag
     3121 cgctggggtc cgctgcctca gtactcccct gacgcgaaag ggacatctgc ccgaacgggt
     3181 tactcaggtc gtcggcctga gagatcatgt ggttattcaa cgggggctgt tgttgagggg
     3241 taggcggtaa agctggcaac tgctgctgtt gttgctgttg ttgttgttgc gccgtacagg
     3301 gcagaagtcc gcgtactgat tgagacgatg ttacctgatt acacacctcg ggagcgccca
     3361 gagccattcc cgggcgggaa atacgcccag cgctcccccc tttgctcgat ccaatagagt
     3421 ccccacgggt aaggatagaa atgccactga aagacgaggc ctttgtcacc ggcgggcgca
     3481 tcgaacggac tgatccgtcg ctatcggtac ttgaccacgg acggggctcc agagatttca
     3541 attcactatc agttgaagat tgacgcgagc tggaggtacg ggataatccc tcgcgattgc
     3601 cgcggaagat ctgacggcgt ttatgagacg acgagctaaa cgcttccttt gataaacgaa
     3661 tatcgttaag gtatccattt tggccagttt ggttatcgtc gcggtccatc gaagcatcgt
     3721 cgcgctttaa aatgaaacgt tgctgaaatt ccgtattttt ctcgtctttg atgtgttcac
     3781 taaagcgctg ttcaggaata cgggtatttg atgtcttgtt aatgattact gcttttccag
     3841 tctggtcccg ggaagcggcc gcacttcccc cggagcctcc gcttccgccc ttgtacagct
     3901 cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc
     3961 gcttctcgtt ggggtctttg ctcagcttgg actgggtgct caggtagtgg ttgtcgggca
     4021 gcagcacggg gccgtcgccg atgggggtgt tctgctggta gtggtcggcg agctgcacgc
     4081 tgccgtcctc gatgttgtgg cggatcttga agttcacctt gatgccgttc ttctgcttgt
     4141 cggccatgat atagacgttg tggctgttgt agttgtactc cagcttgtgc cccaggatgt
     4201 tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg gttcaccagg gtgtcgccct
     4261 cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct
     4321 cctggacgta gccttcgggc atggcggact tgaagaagtc gtgctgcttc atgtggtcgg
     4381 ggtagcggct gaagcactgc acgccgtagg tcagggtggt cacgagggtg ggccagggca
     4441 cgggcagctt gccggtggtg cagatgaact tcagggtcag cttgccgtag gtggcatcgc
     4501 cctcgccctc gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca
     4561 ggatgggcac caccccggtg aacagctcct cgcccttgct caccatggtg gcgaccggtg
     4621 gatcccgggc ccgcggtacc gtcgactgca gaattcggtt cactaaacga gcccggttca
     4681 ctaaacgagc tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca
     4741 atggggcgga gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac
     4801 tcccattgac gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac
     4861 gcccattgat gtactgccaa aaccgcatca ccatggtaat agcgatgact aatacgtaga
     4921 tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc
     4981 atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg
     5041 ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg
     5101 gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc
     5161 agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa
     5221 ctaatgaccc cgtaattgat tactattaat aactagcacg cgttcgaagc ttgagctcgg
     5281 tcccctcggg gttgggaggt gggtctgaaa cgataatggt gaatatccct gcctaactct
     5341 attcactata gaaagtacag caaaaactat tcttaaacct accaagcctc ctactatcat
     5401 tatgaataat tttatatacc acagccaatt tgttatgtta aaccaattcc acaaacttgc
     5461 ccatttatct aattccaata attcttgttc attcttttct tgctggtttt gcgattcttc
     5521 aattaaggag tgtattaagc ttgtgtaatt gttaatttct ctgtcccact ccatccaggt
     5581 cgtgtgattc caaatctgtt ccagagattt attactccaa ctagcattcc aaggcacagc
     5641 agtggtgcaa atgagttttc cagagcaacc ccaaatcccc aggagctgtt gatcctttag
     5701 gtatctttcc acagccagga ttcttgcctg gagctgcttg atgccccaga ctgtgagttg
     5761 caacagatgc tgttgcgcct caatagccct cagcaaattg ttctgctgct gcactatacc
     5821 agacaataat tgtctggcct gtaccgtcag cgtcattgac gctgcgccca tagtgcttcc
     5881 tgctgctccc aagaacccaa ggaacaaagc tcctattccc actgctcttt tttctctctg
     5941 caccactctt ctctttgcct tggtgggtgc tactcctaat ggttcaattt ttactacttt
     6001 atatttatat aattcacttc tccaattgtc cctcatatct cctccttatg gtgttttact
     6061 aaacttttcc atgttctaat cctcatcctg tctacttgcc acacaatcat cacctgccat
     6121 ctgttttcca taatccctaa tgatctttgc ttttcttctt ggcactactt ttatgtcact
     6181 attatcttgt attactactg ccccttcacc tttccagagg agctttgctg gtcctttcca
     6241 aagtggattt ctgctgtccc tgtaataaac ccgaaaattt tgaatttttg taatttgttt
     6301 ttgtaattct ttagtttgta tgtctgttgc tattatgtct actattcttt cccctgcact
     6361 gtacccccca atcccccctt ttcttttaaa attgtggatg aatactgcca tttgtactgc
     6421 tgtcttaaga tgttcagcct gatctcttac ctgtcctata attttcttta attctttatt
     6481 catagattct actactcctt gactttgggg attgtaggga atttccaggt ctgaagatcc
     6541 tgctgcttgc tgtgcttttt tcttactttt gttttgctct tcctctatct tgtctaaagc
     6601 ttccttggtg tcttttatct ctatcctttg atgcacacaa tagagggttg ctactgtatt
     6661 atataatgat ctaagttctt ctgatcctgt ctgaagggat ggttgtagct gtcccagtat
     6721 ttgtctacag ccttctgatg tttctaacag gccagtatta actgcgaatc gttctagctc
     6781 cctgcttgcc catactatat gttttaattt atattttttc tttccccctg gccttaaccg
     6841 aattttttcc catcgcgatc taattctccc ccgcttaata ctgacgctct cgcacccatc
     6901 tctctccttc tagcctccgc tagtcaaaat ttttggcgta ctcaccagtc gccgcccctc
     6961 gcctcttgcc gtgcgcgctt cagcaagccg agtcctgcgt cgagagagct cctctggttt
     7021 ccctttcgct ttcaagtccc tgttcgggcg ccactgctag agattttcca cactgactaa
     7081 aagggtctga gggatctcta gttaccagag tcacacaaca gacgggcaca cactacttga
     7141 agcactcaag gcaagcttta ttgaggctta agcagtgggt tccctagtta gccagagagc
     7201 tcccaggctc agatctggtc taaccagaga gacccagtac aggcaaaaag cagctgctta
     7261 tatgcaggat ctgagggctc gccactcccc agtcccgccc aggccacgcc tcccatggta
     7321 atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac
     7381 tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc
     7441 atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga
     7501 cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat
     7561 gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg
     7621 aactccatat atgggctatg aactaatgac cccgtaattg attactatta ataactagtc
     7681 aataatcaat gtcgacccag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     7741 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     7801 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     7861 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     7921 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     7981 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     8041 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     8101 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     8161 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     8221 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     8281 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     8341 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     8401 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     8461 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     8521 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     8581 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     8641 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     8701 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     8761 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     8821 ctgagcgtca gaccccgtag aaaagatcaa aggatcttc
//
