LOCUS       sequence-488259-        8745 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            Vidr_030, derived from human protein ARI1A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8745)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8745)
  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_030
FEATURES             Location/Qualifiers
     source          1..8745
                     /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(3776..4486,4487..4489,4490..4492))
                     /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(3818..3839)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4167..4186
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4426..4447
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4489..4498
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4574..4777)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4603..4623)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4778..5081
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(5338..5382)
                     /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    5567..5800
                     /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    6160..6277
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6769..6894
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6941..7198
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        7200..7579
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7610..7714
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7715..7783,7784..8575)
                     /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(7933..7952)
                     /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 tgtccttcat tgtaaggcga cgacgctgct
     2101 tcaatacagg ctgtgtagct tcaacggaac ccggcggaaa agtgatgtca cgacggatca
     2161 ttgggggctg gacaggagcc ggcgcaatgt gggatgcagg aacaggaggt ccagccttct
     2221 gcattttcat accagaatgt aaaaacgggc tcttagacgg tgatgtgcgg ttctccatag
     2281 ggcgaggaag aggagcgggg ctactcacct gtgggatatg gttctgcatg ctgggtggtg
     2341 gctgataatt agaaggggga gggcgtgtca taggcgggac cggcgctgac ggtccataag
     2401 gtggttgacg tgtcccatga ctaggccagg aaccctcatg atttgcgcgt tggtcggtat
     2461 gcagcatttc gtctgtgcga ttcacaccgt gatatgctgg tccctggggt gctgatcccg
     2521 tggactggcg gttggcataa ttataggtca tatcgttgcg cccttgccac atggtgccct
     2581 gttgggctac ttccgcggac gcttgaattg gcccgcccat catttggggt ggcatatttt
     2641 gttgtgcgtt cgtgcctggt ggtgctgaca cacgatcgcg cccgaattga aagggaaact
     2701 ggttctgcgg tccgccggcc gggcgacgtt cggtggctgc agtggccgtc gcagggtatg
     2761 cgttaccata ctggttgtaa acatcctgct gaggacttgg ttgggccgct tgttgttggg
     2821 aggcgggctg aggctgagca gggggcagct gttgttgctg cggttgtcct tggccggtag
     2881 aataaggtac actgtacatt tcgccttcat gacgtttagc aggtgggcca taagtgccat
     2941 ccattggacg tttgtagttt tgttgttgtt gttggtacat agtggtttgc tggctcggaa
     3001 aagggcttcc agatggtgtt ccctgggttg agaactgatt tccataggaa tcatgacgtt
     3061 gttgttgctg ctgctgctgt tgtggtgggt aacgactggg agaatacatg ccggaatctg
     3121 gattagaggg cataagattg ggctgaggag ctccagtaga catgtttcct tcgggtccga
     3181 tacctggttc tgtgcggaca cgatcatagg gacccccgta agggtagtgt tggcggggcc
     3241 ccatggcaac atttccaagg ccggggccgg cagcacggga atacgggtct cccatacccc
     3301 cattgggtcc ttgccccgaa ctcatgaaag ggtcgctacc tggcgctttg cgcatcgaac
     3361 cgtatggatc tttgttcggt tcatacgaca tgcgacccat catgtccgaa gtattcatgg
     3421 atggttgata tcccgggttt ggggtcatcg aattacgctt ttgaaacgtg ctatcactac
     3481 catcgttaaa cgcgtcttga atgcctactg aattgctacg gctcataccg ggcaacggcg
     3541 ggatctgtga gtggggggtt gacgcaggcg ttggaggctt caggtcgccc ccttccgcca
     3601 ttgacgaact agtggactgg ggggtttgag gcccttgcat tgaaccgcta ccggcagggg
     3661 acgggggctg aatttttggc tgtgatttct tagagtcggc agcggcaaag atgtcgggag
     3721 gcggatcttc tccccgggaa gcggccgcac ttcccccgga gcctccgctt ccgcccttgt
     3781 acagctcgtc catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt
     3841 gatcgcgctt ctcgttgggg tctttgctca gcttggactg ggtgctcagg tagtggttgt
     3901 cgggcagcag cacggggccg tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct
     3961 gcacgctgcc gtcctcgatg ttgtggcgga tcttgaagtt caccttgatg ccgttcttct
     4021 gcttgtcggc catgatatag acgttgtggc tgttgtagtt gtactccagc ttgtgcccca
     4081 ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc accagggtgt
     4141 cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg
     4201 tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt
     4261 ggtcggggta gcggctgaag cactgcacgc cgtaggtcag ggtggtcacg agggtgggcc
     4321 agggcacggg cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg
     4381 catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg ccgtccagct
     4441 cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc atggtggcga
     4501 ccggtggatc ccgggcccgc ggtaccgtcg actgcagaat tcggttcact aaacgagccc
     4561 ggttcactaa acgagctctg cttatataga cctcccaccg tacacgccta ccgcccattt
     4621 gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa
     4681 acaaactccc attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct
     4741 atccacgccc attgatgtac tgccaaaacc gcatcaccat ggtaatagcg atgactaata
     4801 cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg
     4861 cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat
     4921 gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc
     4981 ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg
     5041 gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc
     5101 tatgaactaa tgaccccgta attgattact attaataact agcacgcgtt cgaagcttga
     5161 gctcggtccc ctcggggttg ggaggtgggt ctgaaacgat aatggtgaat atccctgcct
     5221 aactctattc actatagaaa gtacagcaaa aactattctt aaacctacca agcctcctac
     5281 tatcattatg aataatttta tataccacag ccaatttgtt atgttaaacc aattccacaa
     5341 acttgcccat ttatctaatt ccaataattc ttgttcattc ttttcttgct ggttttgcga
     5401 ttcttcaatt aaggagtgta ttaagcttgt gtaattgtta atttctctgt cccactccat
     5461 ccaggtcgtg tgattccaaa tctgttccag agatttatta ctccaactag cattccaagg
     5521 cacagcagtg gtgcaaatga gttttccaga gcaaccccaa atccccagga gctgttgatc
     5581 ctttaggtat ctttccacag ccaggattct tgcctggagc tgcttgatgc cccagactgt
     5641 gagttgcaac agatgctgtt gcgcctcaat agccctcagc aaattgttct gctgctgcac
     5701 tataccagac aataattgtc tggcctgtac cgtcagcgtc attgacgctg cgcccatagt
     5761 gcttcctgct gctcccaaga acccaaggaa caaagctcct attcccactg ctcttttttc
     5821 tctctgcacc actcttctct ttgccttggt gggtgctact cctaatggtt caatttttac
     5881 tactttatat ttatataatt cacttctcca attgtccctc atatctcctc cttatggtgt
     5941 tttactaaac ttttccatgt tctaatcctc atcctgtcta cttgccacac aatcatcacc
     6001 tgccatctgt tttccataat ccctaatgat ctttgctttt cttcttggca ctacttttat
     6061 gtcactatta tcttgtatta ctactgcccc ttcacctttc cagaggagct ttgctggtcc
     6121 tttccaaagt ggatttctgc tgtccctgta ataaacccga aaattttgaa tttttgtaat
     6181 ttgtttttgt aattctttag tttgtatgtc tgttgctatt atgtctacta ttctttcccc
     6241 tgcactgtac cccccaatcc ccccttttct tttaaaattg tggatgaata ctgccatttg
     6301 tactgctgtc ttaagatgtt cagcctgatc tcttacctgt cctataattt tctttaattc
     6361 tttattcata gattctacta ctccttgact ttggggattg tagggaattt ccaggtctga
     6421 agatcctgct gcttgctgtg cttttttctt acttttgttt tgctcttcct ctatcttgtc
     6481 taaagcttcc ttggtgtctt ttatctctat cctttgatgc acacaataga gggttgctac
     6541 tgtattatat aatgatctaa gttcttctga tcctgtctga agggatggtt gtagctgtcc
     6601 cagtatttgt ctacagcctt ctgatgtttc taacaggcca gtattaactg cgaatcgttc
     6661 tagctccctg cttgcccata ctatatgttt taatttatat tttttctttc cccctggcct
     6721 taaccgaatt ttttcccatc gcgatctaat tctcccccgc ttaatactga cgctctcgca
     6781 cccatctctc tccttctagc ctccgctagt caaaattttt ggcgtactca ccagtcgccg
     6841 cccctcgcct cttgccgtgc gcgcttcagc aagccgagtc ctgcgtcgag agagctcctc
     6901 tggtttccct ttcgctttca agtccctgtt cgggcgccac tgctagagat tttccacact
     6961 gactaaaagg gtctgaggga tctctagtta ccagagtcac acaacagacg ggcacacact
     7021 acttgaagca ctcaaggcaa gctttattga ggcttaagca gtgggttccc tagttagcca
     7081 gagagctccc aggctcagat ctggtctaac cagagagacc cagtacaggc aaaaagcagc
     7141 tgcttatatg caggatctga gggctcgcca ctccccagtc ccgcccaggc cacgcctccc
     7201 atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc
     7261 atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta
     7321 cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc
     7381 cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac
     7441 gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta
     7501 acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa
     7561 ctagtcaata atcaatgtcg acccaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     7621 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     7681 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     7741 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     7801 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     7861 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     7921 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     7981 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     8041 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     8101 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     8161 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     8221 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     8281 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     8341 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     8401 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     8461 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     8521 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     8581 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     8641 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     8701 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttc
//
