LOCUS       sequence-487933-        7344 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_015, derived from human fusion protein ATP5B_VWA3B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7344)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7344)
  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_FOidr_015
FEATURES             Location/Qualifiers
     source          1..7344
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(381..400)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    446..501
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(500..519)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(862..878)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(862..878)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(875..897)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    886..902
                     /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)."
     promoter        complement(join(910..916,917..934,935..940))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    955..976
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1035..1230
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1081..1216
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    1236..1370
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1273..1292)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1327..1346
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1438..1455)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1609..2197)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1689..1708)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2368..3159,3160..3228))
                     /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     2991..3010
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3229..3333)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        3364..3743
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     LTR             3745..4002
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4049..4174
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4666..4783
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5143..5376
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5561..5605
                     /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"
     enhancer        5862..6165
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6166..6369
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6320..6340
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      6445..6454
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6451..6453,6454..6456,6457..7167)
                     /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(6496..6517)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6757..6776)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7104..7125
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 aaaaaaaagg tccttcatgc tgagagtacg aaaacctctt tgttacgttc tcaaatgagt
       61 agtttgcgct catcagcctg ctctgagcgc aaggacggcc tgagcaatgc atcgagtcgt
      121 cgcactgcct gaaccggttc tagataagat tacggtacct ttaagaccaa tgacttacaa
      181 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca
      241 ctcccaaaga agacaagatc tgctttttgc ctgtactggg tctctctggt tagaccagat
      301 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
      361 cagctgctcg agctagcaga tctttttccc tctgccaaaa attatgggga catcatgaag
      421 ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt
      481 tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat
      541 cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac
      601 aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta
      661 ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt
      721 ttttctttaa catccctaaa attttcctta catgttttac tagccagatt tttcctcctc
      781 tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct cgacctgcag
      841 cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
      901 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
      961 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     1021 gtgccagcgg atccgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     1081 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     1141 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     1201 ggcttttttg gaggcctagg cttttgcaaa aagctaactt gtttattgca gcttataatg
     1261 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     1321 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc cgctgcatta
     1381 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     1441 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     1501 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     1561 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1621 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1681 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1741 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1801 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1861 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1921 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     1981 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     2041 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     2101 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     2161 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     2221 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     2281 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     2341 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     2401 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     2461 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     2521 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     2581 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     2641 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     2701 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     2761 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     2821 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     2881 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     2941 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     3001 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     3061 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     3121 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     3181 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     3241 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     3301 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctgg
     3361 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
     3421 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     3481 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     3541 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     3601 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     3661 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     3721 tattagtcat cgctattacc atgggaggcg tggcctgggc gggactgggg agtggcgagc
     3781 cctcagatcc tgcatataag cagctgcttt ttgcctgtac tgggtctctc tggttagacc
     3841 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     3901 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     3961 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga
     4021 cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc
     4081 gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg
     4141 aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc
     4201 gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat
     4261 agtatgggca agcagggagc tagaacgatt cgcagttaat actggcctgt tagaaacatc
     4321 agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga
     4381 acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat
     4441 aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaaaaa
     4501 agcacagcaa gcagcaggat cttcagacct ggaaattccc tacaatcccc aaagtcaagg
     4561 agtagtagaa tctatgaata aagaattaaa gaaaattata ggacaggtaa gagatcaggc
     4621 tgaacatctt aagacagcag tacaaatggc agtattcatc cacaatttta aaagaaaagg
     4681 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     4741 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     4801 cagcagaaat ccactttgga aaggaccagc aaagctcctc tggaaaggtg aaggggcagt
     4861 agtaatacaa gataatagtg acataaaagt agtgccaaga agaaaagcaa agatcattag
     4921 ggattatgga aaacagatgg caggtgatga ttgtgtggca agtagacagg atgaggatta
     4981 gaacatggaa aagtttagta aaacaccata aggaggagat atgagggaca attggagaag
     5041 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     5101 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     5161 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     5221 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     5281 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     5341 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     5401 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     5461 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     5521 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     5581 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     5641 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     5701 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     5761 cccaaccccg aggggaccga gctcaagctt cgaacgcgtg ctagttatta atagtaatca
     5821 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
     5881 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     5941 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     6001 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     6061 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
     6121 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
     6181 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
     6241 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
     6301 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
     6361 agcagagctc gtttagtgaa ccgggctcgt ttagtgaacc gaattctgca gtcgacggta
     6421 ccgcgggccc gggatccacc ggtcgccacc atggtgagca agggcgagga gctgttcacc
     6481 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     6541 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     6601 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     6661 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     6721 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     6781 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     6841 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     6901 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     6961 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     7021 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccaa gctgagcaaa
     7081 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     7141 actctcggca tggacgagct gtacaagggc ggaagcggag gctccggggg aagtgcggcc
     7201 gcttcccggc atcaaaaaga gatttgtagt atgatctcca cccctgaaaa gtgtgcgaag
     7261 ccacagtctg atgttgattc aacgcaaacc tcttcactta acatgcttaa gggtccatgg
     7321 ggactttcag accaaaaagt gcag
//
