LOCUS       sequence-488245-        8253 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            Vidr_015, derived from human protein TBC12.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8253)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8253)
  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_015
FEATURES             Location/Qualifiers
     source          1..8253
                     /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(3284..3994,3995..3997,3998..4000))
                     /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(3326..3347)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3675..3694
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3934..3955
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3997..4006
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4082..4285)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4111..4131)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4286..4589
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(4846..4890)
                     /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    5075..5308
                     /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    5668..5785
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    6277..6402
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6449..6706
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        6708..7087
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7118..7222
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7223..7291,7292..8083)
                     /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(7441..7460)
                     /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 ccggttcacg gctcaaactc aaagttttta cggcgcgaag
     2101 actggggggg tggtttgcaa gtacggcctg tacgcgcctc gtattcttgc tggatgatct
     2161 tgctcgtttt ttgaaggttt tcacggggcg gcaccttacc gaataatttc catcccgggg
     2221 cactgtggac gacagacttc aactctttgg tgcgctttgg gaataaattg cgtgtaaaga
     2281 aatccgcgaa ccctcctgaa cggcgggcgc gcgcacttgc accggcaggg ccttgttcag
     2341 cagctggcaa aggtacgggc gggccagcgg gtggcagatg gtcacgagcc gattggccgc
     2401 gtgaaacttg gaaagtgtta cgactattaa agtgaatatc cgagaagccc agacgtggct
     2461 ctgcacctcc atttgtacgc tcggcggagg ttgctggggg tgcaccttcc tctggaccac
     2521 ctgccccaac tccacgttgc tcggaatcct cggagctact gcaggaagat gcaggtgaat
     2581 ctcccgaatc cgacccggcc ccttccggct cttgagcgtc cgcagcgaca aggcagcatg
     2641 aacgcaatgg atcttccaac ggacttgccc aatctgatgg cgatccggca ccatcagcgt
     2701 cttcgtctcc tggcccttcc aaacgcaagc ggccgtatgg gcggcggcga cgagactcac
     2761 ggcccccagc acgggccaag ccacgtgctt cttccagatc acggcaatca cgaccgggaa
     2821 gaaagccact atcaccattt gtcatccctt cgtggcgtgg agcacgtccg tcggccacct
     2881 ccgccccacg atgcttacta ccactcccca gcaaacacgc atcactgcgt ggtgccgctg
     2941 aaggcggagg ggctccagct tgtcctgcag gcagaggaca atagcaaaga cccggttcta
     3001 actgttcgcc cgctgctgct aaataacgtt gcagaagttg acggggcgga gtttcttcct
     3061 cctcgtcagc ttcttcttct tcatctgctt cctctggtgg ctcaaccgca cccactccgc
     3121 ccccgaagcc tcccgtcgca cgaataacct tacgatcttg tccaactgga tcaggcgcag
     3181 ggaccggcag caatttaggg ttacgacccg aacatgcacc agcgtcttcc ggaccaacca
     3241 tccgggaagc ggccgcactt cccccggagc ctccgcttcc gcccttgtac agctcgtcca
     3301 tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct
     3361 cgttggggtc tttgctcagc ttggactggg tgctcaggta gtggttgtcg ggcagcagca
     3421 cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt
     3481 cctcgatgtt gtggcggatc ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca
     3541 tgatatagac gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt
     3601 cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact
     3661 tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga
     3721 cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc
     3781 ggctgaagca ctgcacgccg taggtcaggg tggtcacgag ggtgggccag ggcacgggca
     3841 gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc
     3901 cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg
     3961 gcaccacccc ggtgaacagc tcctcgccct tgctcaccat ggtggcgacc ggtggatccc
     4021 gggcccgcgg taccgtcgac tgcagaattc ggttcactaa acgagcccgg ttcactaaac
     4081 gagctctgct tatatagacc tcccaccgta cacgcctacc gcccatttgc gtcaatgggg
     4141 cggagttgtt acgacatttt ggaaagtccc gttgattttg gtgccaaaac aaactcccat
     4201 tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc aaaccgctat ccacgcccat
     4261 tgatgtactg ccaaaaccgc atcaccatgg taatagcgat gactaatacg tagatgtact
     4321 gccaagtagg aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac
     4381 cgtcattgac gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt
     4441 gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta
     4501 ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag
     4561 gcgggccatt taccgtaagt tatgtaacgc ggaactccat atatgggcta tgaactaatg
     4621 accccgtaat tgattactat taataactag cacgcgttcg aagcttgagc tcggtcccct
     4681 cggggttggg aggtgggtct gaaacgataa tggtgaatat ccctgcctaa ctctattcac
     4741 tatagaaagt acagcaaaaa ctattcttaa acctaccaag cctcctacta tcattatgaa
     4801 taattttata taccacagcc aatttgttat gttaaaccaa ttccacaaac ttgcccattt
     4861 atctaattcc aataattctt gttcattctt ttcttgctgg ttttgcgatt cttcaattaa
     4921 ggagtgtatt aagcttgtgt aattgttaat ttctctgtcc cactccatcc aggtcgtgtg
     4981 attccaaatc tgttccagag atttattact ccaactagca ttccaaggca cagcagtggt
     5041 gcaaatgagt tttccagagc aaccccaaat ccccaggagc tgttgatcct ttaggtatct
     5101 ttccacagcc aggattcttg cctggagctg cttgatgccc cagactgtga gttgcaacag
     5161 atgctgttgc gcctcaatag ccctcagcaa attgttctgc tgctgcacta taccagacaa
     5221 taattgtctg gcctgtaccg tcagcgtcat tgacgctgcg cccatagtgc ttcctgctgc
     5281 tcccaagaac ccaaggaaca aagctcctat tcccactgct cttttttctc tctgcaccac
     5341 tcttctcttt gccttggtgg gtgctactcc taatggttca atttttacta ctttatattt
     5401 atataattca cttctccaat tgtccctcat atctcctcct tatggtgttt tactaaactt
     5461 ttccatgttc taatcctcat cctgtctact tgccacacaa tcatcacctg ccatctgttt
     5521 tccataatcc ctaatgatct ttgcttttct tcttggcact acttttatgt cactattatc
     5581 ttgtattact actgcccctt cacctttcca gaggagcttt gctggtcctt tccaaagtgg
     5641 atttctgctg tccctgtaat aaacccgaaa attttgaatt tttgtaattt gtttttgtaa
     5701 ttctttagtt tgtatgtctg ttgctattat gtctactatt ctttcccctg cactgtaccc
     5761 cccaatcccc ccttttcttt taaaattgtg gatgaatact gccatttgta ctgctgtctt
     5821 aagatgttca gcctgatctc ttacctgtcc tataattttc tttaattctt tattcataga
     5881 ttctactact ccttgacttt ggggattgta gggaatttcc aggtctgaag atcctgctgc
     5941 ttgctgtgct tttttcttac ttttgttttg ctcttcctct atcttgtcta aagcttcctt
     6001 ggtgtctttt atctctatcc tttgatgcac acaatagagg gttgctactg tattatataa
     6061 tgatctaagt tcttctgatc ctgtctgaag ggatggttgt agctgtccca gtatttgtct
     6121 acagccttct gatgtttcta acaggccagt attaactgcg aatcgttcta gctccctgct
     6181 tgcccatact atatgtttta atttatattt tttctttccc cctggcctta accgaatttt
     6241 ttcccatcgc gatctaattc tcccccgctt aatactgacg ctctcgcacc catctctctc
     6301 cttctagcct ccgctagtca aaatttttgg cgtactcacc agtcgccgcc cctcgcctct
     6361 tgccgtgcgc gcttcagcaa gccgagtcct gcgtcgagag agctcctctg gtttcccttt
     6421 cgctttcaag tccctgttcg ggcgccactg ctagagattt tccacactga ctaaaagggt
     6481 ctgagggatc tctagttacc agagtcacac aacagacggg cacacactac ttgaagcact
     6541 caaggcaagc tttattgagg cttaagcagt gggttcccta gttagccaga gagctcccag
     6601 gctcagatct ggtctaacca gagagaccca gtacaggcaa aaagcagctg cttatatgca
     6661 ggatctgagg gctcgccact ccccagtccc gcccaggcca cgcctcccat ggtaatagcg
     6721 atgactaata cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca
     6781 taatgccagg cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga
     6841 tacacttgat gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa
     6901 tggaaagtcc ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg
     6961 gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc
     7021 atatatgggc tatgaactaa tgaccccgta attgattact attaataact agtcaataat
     7081 caatgtcgac ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     7141 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     7201 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     7261 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     7321 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     7381 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     7441 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     7501 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     7561 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     7621 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     7681 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     7741 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     7801 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     7861 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     7921 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     7981 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     8041 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     8101 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     8161 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     8221 gtcagacccc gtagaaaaga tcaaaggatc ttc
//
