LOCUS       sequence-490780-        8316 bp    DNA     circular SYN 27-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_107, derived from human fusion protein MEF2D_SS18.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8316)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8316)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 27, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: CL20_mEGFP_FOidr_107
FEATURES             Location/Qualifiers
     source          1..8316
                     /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(1849..2559,2560..2562,2563..2565))
                     /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(1891..1912)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2240..2259
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2499..2520
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2562..2571
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2647..2850)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2676..2696)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2851..3154
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3411..3455)
                     /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    3640..3873
                     /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    4233..4350
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4842..4967
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5014..5271
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5273..5652
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5683..5787
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5788..5856,5857..6648)
                     /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(6006..6025)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6819..7407
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7308..7327
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7561..7578
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7646..7780
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7670..7689)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7724..7743
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7786..7981)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7800..7935
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    8040..8061
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8076..8081,8082..8099,8100..8106)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8114..8130
                     /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     8119..8141
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8138..8154
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8138..8154
                     /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 ctgttggtag ttgccgtatt gaccttggtc
      601 gtacccataa gggcgctgct gtggcggctg cggaggtcct ggttgcgtag gacgatagcc
      661 accgtactgt tgtccttggc cttggggata gtttgggtat tgcggtccgg ggccgccttg
      721 agagggccca tagccttgct gttggccagg gtacccctgc tgtccgggat actgctgctg
      781 ctggggtgga tagccctgct gcggtggggg tccctggtac gcatcctgtt gttggccata
      841 ttgtgagtta ccaccctcat agtaatgttg actactatct tcatacgggc ggtcataacc
      901 ctgttccgga taactcggtt gctgataccc gtagtcattg tgaccatcgg ggtagtattg
      961 ttggttcatt ccttccggag gaccctgacc gccatgactg tattgatccc cataatagtc
     1021 ttcttgaccc gaatattgtt gtggagggcc ctgctggggt gggcggtaag gcgggatttg
     1081 gcgttgcccc atcatgtgat taccctgatt gacctgcccc atcatcccca ttgggggctg
     1141 ttgaccctgg tagtgctggc caccaccctg gggcatatta tactgctgac ttgggggctg
     1201 ctgatgcatc atcggaccgg tgttatatgc ggtcggcatt gaagagaacg gtaagccttg
     1261 gctcagaagc gaaggtgttg ccacagaaac gacaggagta gtcaatgaat gcgttgactg
     1321 ggacactcct aagcgctggg cgttgttcag gtccaggtga tcctcagtaa gatggtgcat
     1381 taagcctttt cccgcctgtg aggtgattac acgaaggtcc ggtttacgag agggggcacc
     1441 taactgggtg ctgtgagtgg gcggtggcgg cgacttcgcg ggaatcacct tgttcaggga
     1501 atttccattg gccactggaa gcaatccagg ggacgcgcgc gcactgacat aaccattgcc
     1561 aaccgggctc gggcacgcac cgtttgcgga gttcaaatct cctcccagca tggcaccagc
     1621 agacgcagga cgctgaggaa gtccaggaga cacgctattg cgttgtaaag ccggttgttg
     1681 aggactaagc agacgagggt cggtaagaga tgaggtaacc aaactgggcg ttactaaact
     1741 tccggaggga ttgctgaatt gcaaagatga ttggttactt acaggaactg ttactggcat
     1801 agcgaaccgg gaagcggccg cacttccccc ggagcctccg cttccgccct tgtacagctc
     1861 gtccatgccg agagtgatcc cggcggcggt cacgaactcc agcaggacca tgtgatcgcg
     1921 cttctcgttg gggtctttgc tcagcttgga ctgggtgctc aggtagtggt tgtcgggcag
     1981 cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga gctgcacgct
     2041 gccgtcctcg atgttgtggc ggatcttgaa gttcaccttg atgccgttct tctgcttgtc
     2101 ggccatgata tagacgttgt ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt
     2161 gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc
     2221 gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc
     2281 ctggacgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca tgtggtcggg
     2341 gtagcggctg aagcactgca cgccgtaggt cagggtggtc acgagggtgg gccagggcac
     2401 gggcagcttg ccggtggtgc agatgaactt cagggtcagc ttgccgtagg tggcatcgcc
     2461 ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag
     2521 gatgggcacc accccggtga acagctcctc gcccttgctc accatggtgg cgaccggtgg
     2581 atcccgggcc cgcggtaccg tcgactgcag aattcggttc actaaacgag cccggttcac
     2641 taaacgagct ctgcttatat agacctccca ccgtacacgc ctaccgccca tttgcgtcaa
     2701 tggggcggag ttgttacgac attttggaaa gtcccgttga ttttggtgcc aaaacaaact
     2761 cccattgacg tcaatggggt ggagacttgg aaatccccgt gagtcaaacc gctatccacg
     2821 cccattgatg tactgccaaa accgcatcac catggtaata gcgatgacta atacgtagat
     2881 gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc aggcgggcca
     2941 tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt gatgtactgc
     3001 caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag tccctattgg
     3061 cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt tgggcggtca
     3121 gccaggcggg ccatttaccg taagttatgt aacgcggaac tccatatatg ggctatgaac
     3181 taatgacccc gtaattgatt actattaata actagcacgc gttcgaagct tgagctcggt
     3241 cccctcgggg ttgggaggtg ggtctgaaac gataatggtg aatatccctg cctaactcta
     3301 ttcactatag aaagtacagc aaaaactatt cttaaaccta ccaagcctcc tactatcatt
     3361 atgaataatt ttatatacca cagccaattt gttatgttaa accaattcca caaacttgcc
     3421 catttatcta attccaataa ttcttgttca ttcttttctt gctggttttg cgattcttca
     3481 attaaggagt gtattaagct tgtgtaattg ttaatttctc tgtcccactc catccaggtc
     3541 gtgtgattcc aaatctgttc cagagattta ttactccaac tagcattcca aggcacagca
     3601 gtggtgcaaa tgagttttcc agagcaaccc caaatcccca ggagctgttg atcctttagg
     3661 tatctttcca cagccaggat tcttgcctgg agctgcttga tgccccagac tgtgagttgc
     3721 aacagatgct gttgcgcctc aatagccctc agcaaattgt tctgctgctg cactatacca
     3781 gacaataatt gtctggcctg taccgtcagc gtcattgacg ctgcgcccat agtgcttcct
     3841 gctgctccca agaacccaag gaacaaagct cctattccca ctgctctttt ttctctctgc
     3901 accactcttc tctttgcctt ggtgggtgct actcctaatg gttcaatttt tactacttta
     3961 tatttatata attcacttct ccaattgtcc ctcatatctc ctccttatgg tgttttacta
     4021 aacttttcca tgttctaatc ctcatcctgt ctacttgcca cacaatcatc acctgccatc
     4081 tgttttccat aatccctaat gatctttgct tttcttcttg gcactacttt tatgtcacta
     4141 ttatcttgta ttactactgc cccttcacct ttccagagga gctttgctgg tcctttccaa
     4201 agtggatttc tgctgtccct gtaataaacc cgaaaatttt gaatttttgt aatttgtttt
     4261 tgtaattctt tagtttgtat gtctgttgct attatgtcta ctattctttc ccctgcactg
     4321 taccccccaa tccccccttt tcttttaaaa ttgtggatga atactgccat ttgtactgct
     4381 gtcttaagat gttcagcctg atctcttacc tgtcctataa ttttctttaa ttctttattc
     4441 atagattcta ctactccttg actttgggga ttgtagggaa tttccaggtc tgaagatcct
     4501 gctgcttgct gtgctttttt cttacttttg ttttgctctt cctctatctt gtctaaagct
     4561 tccttggtgt cttttatctc tatcctttga tgcacacaat agagggttgc tactgtatta
     4621 tataatgatc taagttcttc tgatcctgtc tgaagggatg gttgtagctg tcccagtatt
     4681 tgtctacagc cttctgatgt ttctaacagg ccagtattaa ctgcgaatcg ttctagctcc
     4741 ctgcttgccc atactatatg ttttaattta tattttttct ttccccctgg ccttaaccga
     4801 attttttccc atcgcgatct aattctcccc cgcttaatac tgacgctctc gcacccatct
     4861 ctctccttct agcctccgct agtcaaaatt tttggcgtac tcaccagtcg ccgcccctcg
     4921 cctcttgccg tgcgcgcttc agcaagccga gtcctgcgtc gagagagctc ctctggtttc
     4981 cctttcgctt tcaagtccct gttcgggcgc cactgctaga gattttccac actgactaaa
     5041 agggtctgag ggatctctag ttaccagagt cacacaacag acgggcacac actacttgaa
     5101 gcactcaagg caagctttat tgaggcttaa gcagtgggtt ccctagttag ccagagagct
     5161 cccaggctca gatctggtct aaccagagag acccagtaca ggcaaaaagc agctgcttat
     5221 atgcaggatc tgagggctcg ccactcccca gtcccgccca ggccacgcct cccatggtaa
     5281 tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact
     5341 gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc gtacttggca
     5401 tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac
     5461 gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg
     5521 ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacgcgga
     5581 actccatata tgggctatga actaatgacc ccgtaattga ttactattaa taactagtca
     5641 ataatcaatg tcgacccagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     5701 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     5761 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     5821 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     5881 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     5941 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     6001 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     6061 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     6121 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     6181 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     6241 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     6301 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     6361 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     6421 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     6481 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     6541 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     6601 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     6661 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6721 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6781 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6841 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6901 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6961 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     7021 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     7081 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     7141 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     7201 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     7261 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     7321 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     7381 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     7441 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     7501 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     7561 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     7621 cgttggccga ttcattaatg cagcggatcc agacatgata agatacattg atgagtttgg
     7681 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     7741 tgctttattt gtaaccatta taagctgcaa taaacaagtt agctttttgc aaaagcctag
     7801 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg cggcctcggc
     7861 ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg
     7921 agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg
     7981 cggatccgct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     8041 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     8101 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     8161 tacgccaagc ttgggctgca ggtcgaggga tctccataag agaagaggga cagctatgac
     8221 tgggagtagt caggagagga ggaaaaatct ggctagtaaa acatgtaagg aaaattttag
     8281 ggatgttaaa gaaaaaaata acacaaaaca aaatat
//
