LOCUS       sequence-487967-        7392 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_054, derived from human fusion protein EBF1_PDGFRB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7392)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7392)
  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_054
FEATURES             Location/Qualifiers
     source          1..7392
                     /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(925..1635,1636..1638,1639..1641))
                     /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(967..988)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1316..1335
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1575..1596
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1638..1647
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1723..1926)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1752..1772)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1927..2230
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2487..2531)
                     /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    2716..2949
                     /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    3309..3426
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    3918..4043
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4090..4347
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4349..4728
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4759..4863
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4864..4932,4933..5724)
                     /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(5082..5101)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5895..6483
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6384..6403
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6637..6654
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6722..6856
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6746..6765)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6800..6819
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6862..7057)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6876..7011
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7116..7137
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7152..7157,7158..7175,7176..7182)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7190..7206
                     /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     7195..7217
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7214..7230
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7214..7230
                     /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 caggaagcta tcctctgctt ccgcccgagg
      601 cgcagggcac cccgaatccg gcaactgttc cagctctggc tccggctcca cctggagctc
      661 aagctggggc tctggctctg gttcgtcctg gggctccagg gggctgtcac aggagatggt
      721 tgaggaggtg ttgacttcat tcagggtgga gctggctagg ctgggggaac cctccagtgg
      781 gccctcgtca gcaacctcgg gtttggggtc aggcaggggg atgatatagt cgttgtcacc
      841 ctcattgggc tgcacggcag tatagaggac ggagctggtg tcccgggaag cggccgcact
      901 tcccccggag cctccgcttc cgcccttgta cagctcgtcc atgccgagag tgatcccggc
      961 ggcggtcacg aactccagca ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag
     1021 cttggactgg gtgctcaggt agtggttgtc gggcagcagc acggggccgt cgccgatggg
     1081 ggtgttctgc tggtagtggt cggcgagctg cacgctgccg tcctcgatgt tgtggcggat
     1141 cttgaagttc accttgatgc cgttcttctg cttgtcggcc atgatataga cgttgtggct
     1201 gttgtagttg tactccagct tgtgccccag gatgttgccg tcctccttga agtcgatgcc
     1261 cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt
     1321 gtagttgccg tcgtccttga agaagatggt gcgctcctgg acgtagcctt cgggcatggc
     1381 ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag cggctgaagc actgcacgcc
     1441 gtaggtcagg gtggtcacga gggtgggcca gggcacgggc agcttgccgg tggtgcagat
     1501 gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa
     1561 cttgtggccg tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc cggtgaacag
     1621 ctcctcgccc ttgctcacca tggtggcgac cggtggatcc cgggcccgcg gtaccgtcga
     1681 ctgcagaatt cggttcacta aacgagcccg gttcactaaa cgagctctgc ttatatagac
     1741 ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt tacgacattt
     1801 tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggggtggag
     1861 acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact gccaaaaccg
     1921 catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca
     1981 taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg
     2041 gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata
     2101 ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat
     2161 tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag
     2221 ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta
     2281 ttaataacta gcacgcgttc gaagcttgag ctcggtcccc tcggggttgg gaggtgggtc
     2341 tgaaacgata atggtgaata tccctgccta actctattca ctatagaaag tacagcaaaa
     2401 actattctta aacctaccaa gcctcctact atcattatga ataattttat ataccacagc
     2461 caatttgtta tgttaaacca attccacaaa cttgcccatt tatctaattc caataattct
     2521 tgttcattct tttcttgctg gttttgcgat tcttcaatta aggagtgtat taagcttgtg
     2581 taattgttaa tttctctgtc ccactccatc caggtcgtgt gattccaaat ctgttccaga
     2641 gatttattac tccaactagc attccaaggc acagcagtgg tgcaaatgag ttttccagag
     2701 caaccccaaa tccccaggag ctgttgatcc tttaggtatc tttccacagc caggattctt
     2761 gcctggagct gcttgatgcc ccagactgtg agttgcaaca gatgctgttg cgcctcaata
     2821 gccctcagca aattgttctg ctgctgcact ataccagaca ataattgtct ggcctgtacc
     2881 gtcagcgtca ttgacgctgc gcccatagtg cttcctgctg ctcccaagaa cccaaggaac
     2941 aaagctccta ttcccactgc tcttttttct ctctgcacca ctcttctctt tgccttggtg
     3001 ggtgctactc ctaatggttc aatttttact actttatatt tatataattc acttctccaa
     3061 ttgtccctca tatctcctcc ttatggtgtt ttactaaact tttccatgtt ctaatcctca
     3121 tcctgtctac ttgccacaca atcatcacct gccatctgtt ttccataatc cctaatgatc
     3181 tttgcttttc ttcttggcac tacttttatg tcactattat cttgtattac tactgcccct
     3241 tcacctttcc agaggagctt tgctggtcct ttccaaagtg gatttctgct gtccctgtaa
     3301 taaacccgaa aattttgaat ttttgtaatt tgtttttgta attctttagt ttgtatgtct
     3361 gttgctatta tgtctactat tctttcccct gcactgtacc ccccaatccc cccttttctt
     3421 ttaaaattgt ggatgaatac tgccatttgt actgctgtct taagatgttc agcctgatct
     3481 cttacctgtc ctataatttt ctttaattct ttattcatag attctactac tccttgactt
     3541 tggggattgt agggaatttc caggtctgaa gatcctgctg cttgctgtgc ttttttctta
     3601 cttttgtttt gctcttcctc tatcttgtct aaagcttcct tggtgtcttt tatctctatc
     3661 ctttgatgca cacaatagag ggttgctact gtattatata atgatctaag ttcttctgat
     3721 cctgtctgaa gggatggttg tagctgtccc agtatttgtc tacagccttc tgatgtttct
     3781 aacaggccag tattaactgc gaatcgttct agctccctgc ttgcccatac tatatgtttt
     3841 aatttatatt ttttctttcc ccctggcctt aaccgaattt tttcccatcg cgatctaatt
     3901 ctcccccgct taatactgac gctctcgcac ccatctctct ccttctagcc tccgctagtc
     3961 aaaatttttg gcgtactcac cagtcgccgc ccctcgcctc ttgccgtgcg cgcttcagca
     4021 agccgagtcc tgcgtcgaga gagctcctct ggtttccctt tcgctttcaa gtccctgttc
     4081 gggcgccact gctagagatt ttccacactg actaaaaggg tctgagggat ctctagttac
     4141 cagagtcaca caacagacgg gcacacacta cttgaagcac tcaaggcaag ctttattgag
     4201 gcttaagcag tgggttccct agttagccag agagctccca ggctcagatc tggtctaacc
     4261 agagagaccc agtacaggca aaaagcagct gcttatatgc aggatctgag ggctcgccac
     4321 tccccagtcc cgcccaggcc acgcctccca tggtaatagc gatgactaat acgtagatgt
     4381 actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt
     4441 taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca
     4501 agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg
     4561 ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc
     4621 caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta
     4681 atgaccccgt aattgattac tattaataac tagtcaataa tcaatgtcga cccaggtggc
     4741 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     4801 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4861 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     4921 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     4981 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5041 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     5101 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5161 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5221 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5281 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5341 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5401 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5461 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5521 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     5581 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     5641 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     5701 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     5761 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5821 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5881 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5941 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6001 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6061 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6121 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6181 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6241 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6301 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6361 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6421 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6481 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6541 atgttctttc ctgcgttatc ccctgcttct gtggataacc gtattaccgc ctttgagtga
     6601 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     6661 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     6721 ggatccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     6781 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     6841 ctgcaataaa caagttagct ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta
     6901 cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag
     6961 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
     7021 taggggcggg actatggttg ctgactaatt gagatgcgga tccgctggca cgacaggttt
     7081 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     7141 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     7201 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgg gctgcaggtc
     7261 gagggatctc cataagagaa gagggacagc tatgactggg agtagtcagg agaggaggaa
     7321 aaatctggct agtaaaacat gtaaggaaaa ttttagggat gttaaagaaa aaaataacac
     7381 aaaacaaaat at
//
