LOCUS       sequence-487980-        8052 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_071, derived from human fusion protein HEY1_NCOA2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8052)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8052)
  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_071
FEATURES             Location/Qualifiers
     source          1..8052
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     181..200
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    199..254
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     300..319
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(join(1585..2295,2296..2298,2299..2301))
                     /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(1627..1648)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1976..1995
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2235..2256
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2298..2307
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2383..2586)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2412..2432)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2587..2890
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3147..3191)
                     /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    3376..3609
                     /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    3969..4086
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4578..4703
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4750..5007
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5009..5388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5419..5523
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5524..5592,5593..6384)
                     /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(5742..5761)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6555..7143
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7044..7063
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7297..7314
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7382..7516
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7406..7425)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7460..7479
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7522..7717)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7536..7671
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7776..7797
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7812..7817,7818..7835,7836..7842)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7850..7866
                     /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     7855..7877
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7874..7890
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7874..7890
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aaaaaaaatc taacctcaag tcaaggcttt tctatggaat aaggaatgga cagcaggggg
       61 ctgtttcata tactgatgac ctctttatag ccaacctttg ttcatggcag ccagcatatg
      121 ggcatatgtt gccaaactct aaaccaaata ctcattctga tgttttaaat gatttgccct
      181 cccatatgtc cttccgagtg agagacacaa aaaattccaa cacactattg caatgaaaat
      241 aaatttcctt tattagccag aagtcagatg ctcaaggggc ttcatgatgt ccccataatt
      301 tttggcagag ggaaaaagat ctgctagctc gagcagctga agctttattg aggcttaagc
      361 agtgggttcc ctagttagcc agagagctcc caggctcaga tctggtctaa ccagagagac
      421 ccagtacagg caaaaagcag atcttgtctt ctttgggagt gaattagccc ttccagtccc
      481 cccttttctt ttaaaaagtg gctaagatct acagctgcct tgtaagtcat tggtcttaaa
      541 ggtaccgtaa tcttatctag aaccggttca ctggtccagg agagctccct catcactcgg
      601 agactcagct gcaggatgtg gacatagcaa gtcatctgga gaactgccaa atggctgcct
      661 gttttggctg gcaaaagacg cctggtctat aggcaggatg ctttcaggcc atggggcagt
      721 ctgatttgga ggagcttgtt gttggctata ctgaggtccc cccatgttca tctctaattc
      781 agatggccct atattcatga cctgagactg aagcgtctgt ctttggccag gctggctgct
      841 gggcctcatg gggatgctgg ctgctgggtt ccgaatcata cctccttgga ctggccggtt
      901 catggcactg gtggtagcag cacaggtgac tctcacagcc gaactctgcg gtgcccattc
      961 tccagatggc atagtaggcc gagaagcact gttaccaatc attcctgtgc tactgttccc
     1021 taaatttcct tggtttccta tcatcccttg attacccatc attcctggct gaggtatcac
     1081 tgagtaggga ctactgtttc tgattggtgg gaaaggtcca gcaccagttg ggctttgcaa
     1141 tgtgatgtca agtggtaaat tctggtttgg caataacctg cccagttgcc ctggtcgtgg
     1201 gttattaaaa gctttccctc ctgccgtatg cagcattttc aggtgatcca cggtcatctg
     1261 caggatctcg gctttttcta gcttagcaga tccctgcttc tcaaaagcac tgggtaccag
     1321 ccttctcagc tcagacaaac tgttattgat ccggtctcgt cggcgcttct caattattcc
     1381 tctccgtctt tttctggcca aaatctggga agatgtagtt ggggacatgg aacctagagc
     1441 cgaactcaag tttccattct cgtccgcact ctccttctcc acctcgatgg tctcgtccag
     1501 ctcgctgtcc gaggagctgt actcggggtg agctcgcttc atccgggaag cggccgcact
     1561 tcccccggag cctccgcttc cgcccttgta cagctcgtcc atgccgagag tgatcccggc
     1621 ggcggtcacg aactccagca ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag
     1681 cttggactgg gtgctcaggt agtggttgtc gggcagcagc acggggccgt cgccgatggg
     1741 ggtgttctgc tggtagtggt cggcgagctg cacgctgccg tcctcgatgt tgtggcggat
     1801 cttgaagttc accttgatgc cgttcttctg cttgtcggcc atgatataga cgttgtggct
     1861 gttgtagttg tactccagct tgtgccccag gatgttgccg tcctccttga agtcgatgcc
     1921 cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt
     1981 gtagttgccg tcgtccttga agaagatggt gcgctcctgg acgtagcctt cgggcatggc
     2041 ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag cggctgaagc actgcacgcc
     2101 gtaggtcagg gtggtcacga gggtgggcca gggcacgggc agcttgccgg tggtgcagat
     2161 gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa
     2221 cttgtggccg tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc cggtgaacag
     2281 ctcctcgccc ttgctcacca tggtggcgac cggtggatcc cgggcccgcg gtaccgtcga
     2341 ctgcagaatt cggttcacta aacgagcccg gttcactaaa cgagctctgc ttatatagac
     2401 ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt tacgacattt
     2461 tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggggtggag
     2521 acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact gccaaaaccg
     2581 catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca
     2641 taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg
     2701 gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata
     2761 ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat
     2821 tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag
     2881 ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta
     2941 ttaataacta gcacgcgttc gaagcttgag ctcggtcccc tcggggttgg gaggtgggtc
     3001 tgaaacgata atggtgaata tccctgccta actctattca ctatagaaag tacagcaaaa
     3061 actattctta aacctaccaa gcctcctact atcattatga ataattttat ataccacagc
     3121 caatttgtta tgttaaacca attccacaaa cttgcccatt tatctaattc caataattct
     3181 tgttcattct tttcttgctg gttttgcgat tcttcaatta aggagtgtat taagcttgtg
     3241 taattgttaa tttctctgtc ccactccatc caggtcgtgt gattccaaat ctgttccaga
     3301 gatttattac tccaactagc attccaaggc acagcagtgg tgcaaatgag ttttccagag
     3361 caaccccaaa tccccaggag ctgttgatcc tttaggtatc tttccacagc caggattctt
     3421 gcctggagct gcttgatgcc ccagactgtg agttgcaaca gatgctgttg cgcctcaata
     3481 gccctcagca aattgttctg ctgctgcact ataccagaca ataattgtct ggcctgtacc
     3541 gtcagcgtca ttgacgctgc gcccatagtg cttcctgctg ctcccaagaa cccaaggaac
     3601 aaagctccta ttcccactgc tcttttttct ctctgcacca ctcttctctt tgccttggtg
     3661 ggtgctactc ctaatggttc aatttttact actttatatt tatataattc acttctccaa
     3721 ttgtccctca tatctcctcc ttatggtgtt ttactaaact tttccatgtt ctaatcctca
     3781 tcctgtctac ttgccacaca atcatcacct gccatctgtt ttccataatc cctaatgatc
     3841 tttgcttttc ttcttggcac tacttttatg tcactattat cttgtattac tactgcccct
     3901 tcacctttcc agaggagctt tgctggtcct ttccaaagtg gatttctgct gtccctgtaa
     3961 taaacccgaa aattttgaat ttttgtaatt tgtttttgta attctttagt ttgtatgtct
     4021 gttgctatta tgtctactat tctttcccct gcactgtacc ccccaatccc cccttttctt
     4081 ttaaaattgt ggatgaatac tgccatttgt actgctgtct taagatgttc agcctgatct
     4141 cttacctgtc ctataatttt ctttaattct ttattcatag attctactac tccttgactt
     4201 tggggattgt agggaatttc caggtctgaa gatcctgctg cttgctgtgc ttttttctta
     4261 cttttgtttt gctcttcctc tatcttgtct aaagcttcct tggtgtcttt tatctctatc
     4321 ctttgatgca cacaatagag ggttgctact gtattatata atgatctaag ttcttctgat
     4381 cctgtctgaa gggatggttg tagctgtccc agtatttgtc tacagccttc tgatgtttct
     4441 aacaggccag tattaactgc gaatcgttct agctccctgc ttgcccatac tatatgtttt
     4501 aatttatatt ttttctttcc ccctggcctt aaccgaattt tttcccatcg cgatctaatt
     4561 ctcccccgct taatactgac gctctcgcac ccatctctct ccttctagcc tccgctagtc
     4621 aaaatttttg gcgtactcac cagtcgccgc ccctcgcctc ttgccgtgcg cgcttcagca
     4681 agccgagtcc tgcgtcgaga gagctcctct ggtttccctt tcgctttcaa gtccctgttc
     4741 gggcgccact gctagagatt ttccacactg actaaaaggg tctgagggat ctctagttac
     4801 cagagtcaca caacagacgg gcacacacta cttgaagcac tcaaggcaag ctttattgag
     4861 gcttaagcag tgggttccct agttagccag agagctccca ggctcagatc tggtctaacc
     4921 agagagaccc agtacaggca aaaagcagct gcttatatgc aggatctgag ggctcgccac
     4981 tccccagtcc cgcccaggcc acgcctccca tggtaatagc gatgactaat acgtagatgt
     5041 actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt
     5101 taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca
     5161 agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg
     5221 ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc
     5281 caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta
     5341 atgaccccgt aattgattac tattaataac tagtcaataa tcaatgtcga cccaggtggc
     5401 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5461 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5521 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     5581 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5641 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5701 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     5761 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5821 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5881 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5941 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     6001 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     6061 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     6121 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     6181 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     6241 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     6301 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     6361 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6421 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6481 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6541 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6601 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6661 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6721 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6781 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6841 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6901 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6961 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     7021 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     7081 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     7141 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     7201 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     7261 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     7321 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     7381 ggatccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     7441 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     7501 ctgcaataaa caagttagct ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta
     7561 cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag
     7621 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
     7681 taggggcggg actatggttg ctgactaatt gagatgcgga tccgctggca cgacaggttt
     7741 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     7801 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     7861 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgg gctgcaggtc
     7921 gagggatctc cataagagaa gagggacagc tatgactggg agtagtcagg agaggaggaa
     7981 aaatctggct agtaaaacat gtaaggaaaa ttttagggat gttaaagaaa aaaataacac
     8041 aaaacaaaat at
//
