LOCUS       sequence-488247-        8811 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            Vidr_018, derived from human protein P121A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8811)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8811)
  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_018
FEATURES             Location/Qualifiers
     source          1..8811
                     /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(3842..4552,4553..4555,4556..4558))
                     /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(3884..3905)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4233..4252
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4492..4513
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4555..4564
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4640..4843)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4669..4689)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4844..5147
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(5404..5448)
                     /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    5633..5866
                     /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    6226..6343
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6835..6960
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             7007..7264
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        7266..7645
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7676..7780
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7781..7849,7850..8641)
                     /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(7999..8018)
                     /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 ctggcttcga agcgctatcg gtcgaagggc
     2101 tggctgaggt aatcggggca accgccgagg tggccgatgc taaaccagta aaaagaggag
     2161 cggtagtcgt tggggccgtc gcaggtgtcg tagtttgctt gaagaagggt gccggcaacg
     2221 gaactgatgc gggaggcccc attgatgaga agactggttg gaatgtaggc gctgtcgtgg
     2281 aagtagtagt gggcagagac gaactagagg gtgcagtggc cgttacgctg ggtcctggtg
     2341 gcgttgggcc ctctttctct gacttaggtg gtgcggtgaa aattggtttg aacattggcg
     2401 gtgccgagct ggctgcgggg gcggctggcg aggaggggct ggtattctgt gtgccaaaaa
     2461 ggaaggattg cttaggcgct ggagaggggg ctgatgtggc ttggggtttt gtcgcagttt
     2521 ctgcctgcaa cgtagggggc gcttttgtat cagtagcagg caccatactt ggagcaggaa
     2581 ttaagcctaa taaagtcgtt ggcggtttag agtcgaagct tggggaaggt aataatccgg
     2641 gaggcccgga ctgggaaagg cccaggggag gcagtaaact gggggtctta ggcggactaa
     2701 gtgcttcggt agttgctgcc ccggcgcttt cgggacacgg cggcagtgat gggggagtct
     2761 gcattttctt caatgattcc agaagcgggt ttgtcgaggg agcaagtaac gacgtaggtg
     2821 gcgaggcagg tgcagcggct ggaagagtaa aagtaaatga tggttgcgtg ataggcgggg
     2881 tttctgttac actgttagac gcagcgtcac tcttatcttc cagagcctga ttgaaccatt
     2941 gtaacgaagc cttcttctca aggtccaagt cctcagctgt gatactgtat cccagctggg
     3001 gcgggggcgg taaggttaac tgttcaccgc ggcgagacgg cagcaattgt accttacgtt
     3061 tacgctgtcc ggagcttcct ggcgtactct gtgaattcga gttttgtttc ttacgaggcg
     3121 ttgtgtccgc ggccttctca ccttggcttt cgcgatccgc ggccaggggg gtggacgaac
     3181 tactatggtg gcacaactcc tcctcacgga tcttttttgc tgggcgctcg ggagtttgcg
     3241 agcgcgatga tgctgggcta gagaagggcg aagaactggg gccgttgcgt ttccacagct
     3301 gcgagatccc gcgagtagag ctgtagctgg atgtgattgc gttacgagac gagctaggga
     3361 taccggaggc atacgcacct gtcaaacttg acatggaact tgagcgggag cgcttattca
     3421 ggtgatcgtc tgaactttgg gaattcaatc cacgcttaag actccctggc ttgggcacaa
     3481 aggaagcggg tacaccatta gcgaccaaag gttcgaaagc cgaatggcca ctgcccgacg
     3541 agtcgtgacg gcggcgcttg ttttcttggc catctaagaa gatctggtct tcttcctcta
     3601 ccgtacgttt tttttccttc tctttaagag cagacagcac agtctcttta gcgcacgggt
     3661 caggcgcgtt gctactaggg gaactcaagg tgctcgaaat aatctgttcg ggaattgctg
     3721 aacgggaaaa acggcggtct ggtggcgcaa tgcgaacagt tacaggcgaa caaaccatac
     3781 gtgagttgcg cggagaaagc cgggaagcgg ccgcacttcc cccggagcct ccgcttccgc
     3841 ccttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga
     3901 ccatgtgatc gcgcttctcg ttggggtctt tgctcagctt ggactgggtg ctcaggtagt
     3961 ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg
     4021 cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc ttgatgccgt
     4081 tcttctgctt gtcggccatg atatagacgt tgtggctgtt gtagttgtac tccagcttgt
     4141 gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca
     4201 gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga
     4261 agatggtgcg ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct
     4321 tcatgtggtc ggggtagcgg ctgaagcact gcacgccgta ggtcagggtg gtcacgaggg
     4381 tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt
     4441 aggtggcatc gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt
     4501 ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatgg
     4561 tggcgaccgg tggatcccgg gcccgcggta ccgtcgactg cagaattcgg ttcactaaac
     4621 gagcccggtt cactaaacga gctctgctta tatagacctc ccaccgtaca cgcctaccgc
     4681 ccatttgcgt caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt
     4741 gccaaaacaa actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa
     4801 accgctatcc acgcccattg atgtactgcc aaaaccgcat caccatggta atagcgatga
     4861 ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat
     4921 gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca
     4981 cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga
     5041 aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt
     5101 cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat
     5161 atgggctatg aactaatgac cccgtaattg attactatta ataactagca cgcgttcgaa
     5221 gcttgagctc ggtcccctcg gggttgggag gtgggtctga aacgataatg gtgaatatcc
     5281 ctgcctaact ctattcacta tagaaagtac agcaaaaact attcttaaac ctaccaagcc
     5341 tcctactatc attatgaata attttatata ccacagccaa tttgttatgt taaaccaatt
     5401 ccacaaactt gcccatttat ctaattccaa taattcttgt tcattctttt cttgctggtt
     5461 ttgcgattct tcaattaagg agtgtattaa gcttgtgtaa ttgttaattt ctctgtccca
     5521 ctccatccag gtcgtgtgat tccaaatctg ttccagagat ttattactcc aactagcatt
     5581 ccaaggcaca gcagtggtgc aaatgagttt tccagagcaa ccccaaatcc ccaggagctg
     5641 ttgatccttt aggtatcttt ccacagccag gattcttgcc tggagctgct tgatgcccca
     5701 gactgtgagt tgcaacagat gctgttgcgc ctcaatagcc ctcagcaaat tgttctgctg
     5761 ctgcactata ccagacaata attgtctggc ctgtaccgtc agcgtcattg acgctgcgcc
     5821 catagtgctt cctgctgctc ccaagaaccc aaggaacaaa gctcctattc ccactgctct
     5881 tttttctctc tgcaccactc ttctctttgc cttggtgggt gctactccta atggttcaat
     5941 ttttactact ttatatttat ataattcact tctccaattg tccctcatat ctcctcctta
     6001 tggtgtttta ctaaactttt ccatgttcta atcctcatcc tgtctacttg ccacacaatc
     6061 atcacctgcc atctgttttc cataatccct aatgatcttt gcttttcttc ttggcactac
     6121 ttttatgtca ctattatctt gtattactac tgccccttca cctttccaga ggagctttgc
     6181 tggtcctttc caaagtggat ttctgctgtc cctgtaataa acccgaaaat tttgaatttt
     6241 tgtaatttgt ttttgtaatt ctttagtttg tatgtctgtt gctattatgt ctactattct
     6301 ttcccctgca ctgtaccccc caatcccccc ttttctttta aaattgtgga tgaatactgc
     6361 catttgtact gctgtcttaa gatgttcagc ctgatctctt acctgtccta taattttctt
     6421 taattcttta ttcatagatt ctactactcc ttgactttgg ggattgtagg gaatttccag
     6481 gtctgaagat cctgctgctt gctgtgcttt tttcttactt ttgttttgct cttcctctat
     6541 cttgtctaaa gcttccttgg tgtcttttat ctctatcctt tgatgcacac aatagagggt
     6601 tgctactgta ttatataatg atctaagttc ttctgatcct gtctgaaggg atggttgtag
     6661 ctgtcccagt atttgtctac agccttctga tgtttctaac aggccagtat taactgcgaa
     6721 tcgttctagc tccctgcttg cccatactat atgttttaat ttatattttt tctttccccc
     6781 tggccttaac cgaatttttt cccatcgcga tctaattctc ccccgcttaa tactgacgct
     6841 ctcgcaccca tctctctcct tctagcctcc gctagtcaaa atttttggcg tactcaccag
     6901 tcgccgcccc tcgcctcttg ccgtgcgcgc ttcagcaagc cgagtcctgc gtcgagagag
     6961 ctcctctggt ttccctttcg ctttcaagtc cctgttcggg cgccactgct agagattttc
     7021 cacactgact aaaagggtct gagggatctc tagttaccag agtcacacaa cagacgggca
     7081 cacactactt gaagcactca aggcaagctt tattgaggct taagcagtgg gttccctagt
     7141 tagccagaga gctcccaggc tcagatctgg tctaaccaga gagacccagt acaggcaaaa
     7201 agcagctgct tatatgcagg atctgagggc tcgccactcc ccagtcccgc ccaggccacg
     7261 cctcccatgg taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat
     7321 aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg
     7381 ggcgtacttg gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac
     7441 tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt
     7501 attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt
     7561 tatgtaacgc ggaactccat atatgggcta tgaactaatg accccgtaat tgattactat
     7621 taataactag tcaataatca atgtcgaccc aggtggcact tttcggggaa atgtgcgcgg
     7681 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     7741 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     7801 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     7861 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     7921 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     7981 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     8041 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     8101 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     8161 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     8221 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     8281 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     8341 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     8401 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     8461 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     8521 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     8581 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     8641 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     8701 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     8761 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt c
//
