LOCUS       sequence-488135-        7476 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_112, derived from human fusion protein MYDGF_DNM2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7476)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7476)
  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_112
FEATURES             Location/Qualifiers
     source          1..7476
                     /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(1009..1719,1720..1722,1723..1725))
                     /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(1051..1072)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1400..1419
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1659..1680
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1722..1731
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1807..2010)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1836..1856)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2011..2314
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2571..2615)
                     /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    2800..3033
                     /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    3393..3510
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4002..4127
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4174..4431
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4433..4812
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4843..4947
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4948..5016,5017..5808)
                     /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(5166..5185)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5979..6567
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6468..6487
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6721..6738
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6806..6940
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6830..6849)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6884..6903
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6946..7141)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6960..7095
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7200..7221
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7236..7241,7242..7259,7260..7266)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7274..7290
                     /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     7279..7301
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7298..7314
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7298..7314
                     /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 gtcaagaagt gagggctctg ctggacggat
      601 aatggttggg cgggatggag ctgccggcgg acgacggcta ggaacgccgg gtggaatacc
      661 gggtggaata cggacggggc gacttggaat ttgtggcggt gcagggaaaa ggtccgaatt
      721 ggcaaagaca ctctgtggtc ccggacgtga gggaataggg ggtgcactaa aactagccgc
      781 tgcccctacg gggaccggga taagaggagg accgggagtt ggaccgcgca cggccggtgg
      841 acgtccaggt gggtgaatag aagagacggg gtggcgctgg ggagtaggag agtgagaaga
      901 ggcactttgt aaccaagtat catcgaccgg aggaggcaca ggtgtgctca cagtactcgt
      961 agagatccgg gaagcggccg cacttccccc ggagcctccg cttccgccct tgtacagctc
     1021 gtccatgccg agagtgatcc cggcggcggt cacgaactcc agcaggacca tgtgatcgcg
     1081 cttctcgttg gggtctttgc tcagcttgga ctgggtgctc aggtagtggt tgtcgggcag
     1141 cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga gctgcacgct
     1201 gccgtcctcg atgttgtggc ggatcttgaa gttcaccttg atgccgttct tctgcttgtc
     1261 ggccatgata tagacgttgt ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt
     1321 gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc
     1381 gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc
     1441 ctggacgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca tgtggtcggg
     1501 gtagcggctg aagcactgca cgccgtaggt cagggtggtc acgagggtgg gccagggcac
     1561 gggcagcttg ccggtggtgc agatgaactt cagggtcagc ttgccgtagg tggcatcgcc
     1621 ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag
     1681 gatgggcacc accccggtga acagctcctc gcccttgctc accatggtgg cgaccggtgg
     1741 atcccgggcc cgcggtaccg tcgactgcag aattcggttc actaaacgag cccggttcac
     1801 taaacgagct ctgcttatat agacctccca ccgtacacgc ctaccgccca tttgcgtcaa
     1861 tggggcggag ttgttacgac attttggaaa gtcccgttga ttttggtgcc aaaacaaact
     1921 cccattgacg tcaatggggt ggagacttgg aaatccccgt gagtcaaacc gctatccacg
     1981 cccattgatg tactgccaaa accgcatcac catggtaata gcgatgacta atacgtagat
     2041 gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc aggcgggcca
     2101 tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt gatgtactgc
     2161 caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag tccctattgg
     2221 cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt tgggcggtca
     2281 gccaggcggg ccatttaccg taagttatgt aacgcggaac tccatatatg ggctatgaac
     2341 taatgacccc gtaattgatt actattaata actagcacgc gttcgaagct tgagctcggt
     2401 cccctcgggg ttgggaggtg ggtctgaaac gataatggtg aatatccctg cctaactcta
     2461 ttcactatag aaagtacagc aaaaactatt cttaaaccta ccaagcctcc tactatcatt
     2521 atgaataatt ttatatacca cagccaattt gttatgttaa accaattcca caaacttgcc
     2581 catttatcta attccaataa ttcttgttca ttcttttctt gctggttttg cgattcttca
     2641 attaaggagt gtattaagct tgtgtaattg ttaatttctc tgtcccactc catccaggtc
     2701 gtgtgattcc aaatctgttc cagagattta ttactccaac tagcattcca aggcacagca
     2761 gtggtgcaaa tgagttttcc agagcaaccc caaatcccca ggagctgttg atcctttagg
     2821 tatctttcca cagccaggat tcttgcctgg agctgcttga tgccccagac tgtgagttgc
     2881 aacagatgct gttgcgcctc aatagccctc agcaaattgt tctgctgctg cactatacca
     2941 gacaataatt gtctggcctg taccgtcagc gtcattgacg ctgcgcccat agtgcttcct
     3001 gctgctccca agaacccaag gaacaaagct cctattccca ctgctctttt ttctctctgc
     3061 accactcttc tctttgcctt ggtgggtgct actcctaatg gttcaatttt tactacttta
     3121 tatttatata attcacttct ccaattgtcc ctcatatctc ctccttatgg tgttttacta
     3181 aacttttcca tgttctaatc ctcatcctgt ctacttgcca cacaatcatc acctgccatc
     3241 tgttttccat aatccctaat gatctttgct tttcttcttg gcactacttt tatgtcacta
     3301 ttatcttgta ttactactgc cccttcacct ttccagagga gctttgctgg tcctttccaa
     3361 agtggatttc tgctgtccct gtaataaacc cgaaaatttt gaatttttgt aatttgtttt
     3421 tgtaattctt tagtttgtat gtctgttgct attatgtcta ctattctttc ccctgcactg
     3481 taccccccaa tccccccttt tcttttaaaa ttgtggatga atactgccat ttgtactgct
     3541 gtcttaagat gttcagcctg atctcttacc tgtcctataa ttttctttaa ttctttattc
     3601 atagattcta ctactccttg actttgggga ttgtagggaa tttccaggtc tgaagatcct
     3661 gctgcttgct gtgctttttt cttacttttg ttttgctctt cctctatctt gtctaaagct
     3721 tccttggtgt cttttatctc tatcctttga tgcacacaat agagggttgc tactgtatta
     3781 tataatgatc taagttcttc tgatcctgtc tgaagggatg gttgtagctg tcccagtatt
     3841 tgtctacagc cttctgatgt ttctaacagg ccagtattaa ctgcgaatcg ttctagctcc
     3901 ctgcttgccc atactatatg ttttaattta tattttttct ttccccctgg ccttaaccga
     3961 attttttccc atcgcgatct aattctcccc cgcttaatac tgacgctctc gcacccatct
     4021 ctctccttct agcctccgct agtcaaaatt tttggcgtac tcaccagtcg ccgcccctcg
     4081 cctcttgccg tgcgcgcttc agcaagccga gtcctgcgtc gagagagctc ctctggtttc
     4141 cctttcgctt tcaagtccct gttcgggcgc cactgctaga gattttccac actgactaaa
     4201 agggtctgag ggatctctag ttaccagagt cacacaacag acgggcacac actacttgaa
     4261 gcactcaagg caagctttat tgaggcttaa gcagtgggtt ccctagttag ccagagagct
     4321 cccaggctca gatctggtct aaccagagag acccagtaca ggcaaaaagc agctgcttat
     4381 atgcaggatc tgagggctcg ccactcccca gtcccgccca ggccacgcct cccatggtaa
     4441 tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact
     4501 gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc gtacttggca
     4561 tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac
     4621 gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg
     4681 ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacgcgga
     4741 actccatata tgggctatga actaatgacc ccgtaattga ttactattaa taactagtca
     4801 ataatcaatg tcgacccagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4861 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4921 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     4981 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     5041 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     5101 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     5161 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     5221 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     5281 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5341 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     5401 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5461 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     5521 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     5581 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5641 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     5701 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5761 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     5821 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5881 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5941 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6001 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6061 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6121 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6181 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6241 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6301 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6361 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6421 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6481 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6541 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6601 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6661 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     6721 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     6781 cgttggccga ttcattaatg cagcggatcc agacatgata agatacattg atgagtttgg
     6841 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     6901 tgctttattt gtaaccatta taagctgcaa taaacaagtt agctttttgc aaaagcctag
     6961 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg cggcctcggc
     7021 ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg
     7081 agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg
     7141 cggatccgct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     7201 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     7261 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     7321 tacgccaagc ttgggctgca ggtcgaggga tctccataag agaagaggga cagctatgac
     7381 tgggagtagt caggagagga ggaaaaatct ggctagtaaa acatgtaagg aaaattttag
     7441 ggatgttaaa gaaaaaaata acacaaaaca aaatat
//
