LOCUS       sequence-487956-        7404 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_042, derived from human fusion protein CSF2RA_CRLF2; 
            IGH_CRLF2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7404)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7404)
  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_042
FEATURES             Location/Qualifiers
     source          1..7404
                     /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(937..1647,1648..1650,1651..1653))
                     /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(979..1000)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1328..1347
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1587..1608
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1650..1659
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1735..1938)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1764..1784)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1939..2242
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2499..2543)
                     /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    2728..2961
                     /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    3321..3438
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    3930..4055
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4102..4359
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4361..4740
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4771..4875
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4876..4944,4945..5736)
                     /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(5094..5113)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5907..6495
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6396..6415
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6649..6666
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6734..6868
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6758..6777)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6812..6831
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6874..7069)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6888..7023
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7128..7149
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7164..7169,7170..7187,7188..7194)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7202..7218
                     /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     7207..7229
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7226..7242
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7226..7242
                     /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 aagggccacg tacgagcgat cattcatcac
      601 gaaagtgaac cccccgatgg ttactacatc tccgccttgc aacggttggt ggggcaattg
      661 caacgacccg ccgctggctt ccttttcttc tgtttggggg tccaacatgc gaggactttc
      721 agcttcagtc tttgccagtt ggacgaccaa aggttcctca gggcccgact cctgttcggc
      781 gcccgccatc ttgtgcagat gggccacatt ctgtgtatcg gtaatccatt cttgaaagtt
      841 tccctggtga atctcgaaca ggccagggaa gatacttttt ggatcaggta cgctccggga
      901 agcggccgca cttcccccgg agcctccgct tccgcccttg tacagctcgt ccatgccgag
      961 agtgatcccg gcggcggtca cgaactccag caggaccatg tgatcgcgct tctcgttggg
     1021 gtctttgctc agcttggact gggtgctcag gtagtggttg tcgggcagca gcacggggcc
     1081 gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat
     1141 gttgtggcgg atcttgaagt tcaccttgat gccgttcttc tgcttgtcgg ccatgatata
     1201 gacgttgtgg ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt
     1261 gaagtcgatg cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc
     1321 ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc
     1381 ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa
     1441 gcactgcacg ccgtaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc
     1501 ggtggtgcag atgaacttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc
     1561 ggacacgctg aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac
     1621 cccggtgaac agctcctcgc ccttgctcac catggtggcg accggtggat cccgggcccg
     1681 cggtaccgtc gactgcagaa ttcggttcac taaacgagcc cggttcacta aacgagctct
     1741 gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt
     1801 gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc
     1861 aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta
     1921 ctgccaaaac cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt
     1981 aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt
     2041 gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt
     2101 ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg
     2161 aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc
     2221 atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt
     2281 aattgattac tattaataac tagcacgcgt tcgaagcttg agctcggtcc cctcggggtt
     2341 gggaggtggg tctgaaacga taatggtgaa tatccctgcc taactctatt cactatagaa
     2401 agtacagcaa aaactattct taaacctacc aagcctccta ctatcattat gaataatttt
     2461 atataccaca gccaatttgt tatgttaaac caattccaca aacttgccca tttatctaat
     2521 tccaataatt cttgttcatt cttttcttgc tggttttgcg attcttcaat taaggagtgt
     2581 attaagcttg tgtaattgtt aatttctctg tcccactcca tccaggtcgt gtgattccaa
     2641 atctgttcca gagatttatt actccaacta gcattccaag gcacagcagt ggtgcaaatg
     2701 agttttccag agcaacccca aatccccagg agctgttgat cctttaggta tctttccaca
     2761 gccaggattc ttgcctggag ctgcttgatg ccccagactg tgagttgcaa cagatgctgt
     2821 tgcgcctcaa tagccctcag caaattgttc tgctgctgca ctataccaga caataattgt
     2881 ctggcctgta ccgtcagcgt cattgacgct gcgcccatag tgcttcctgc tgctcccaag
     2941 aacccaagga acaaagctcc tattcccact gctctttttt ctctctgcac cactcttctc
     3001 tttgccttgg tgggtgctac tcctaatggt tcaattttta ctactttata tttatataat
     3061 tcacttctcc aattgtccct catatctcct ccttatggtg ttttactaaa cttttccatg
     3121 ttctaatcct catcctgtct acttgccaca caatcatcac ctgccatctg ttttccataa
     3181 tccctaatga tctttgcttt tcttcttggc actactttta tgtcactatt atcttgtatt
     3241 actactgccc cttcaccttt ccagaggagc tttgctggtc ctttccaaag tggatttctg
     3301 ctgtccctgt aataaacccg aaaattttga atttttgtaa tttgtttttg taattcttta
     3361 gtttgtatgt ctgttgctat tatgtctact attctttccc ctgcactgta ccccccaatc
     3421 cccccttttc ttttaaaatt gtggatgaat actgccattt gtactgctgt cttaagatgt
     3481 tcagcctgat ctcttacctg tcctataatt ttctttaatt ctttattcat agattctact
     3541 actccttgac tttggggatt gtagggaatt tccaggtctg aagatcctgc tgcttgctgt
     3601 gcttttttct tacttttgtt ttgctcttcc tctatcttgt ctaaagcttc cttggtgtct
     3661 tttatctcta tcctttgatg cacacaatag agggttgcta ctgtattata taatgatcta
     3721 agttcttctg atcctgtctg aagggatggt tgtagctgtc ccagtatttg tctacagcct
     3781 tctgatgttt ctaacaggcc agtattaact gcgaatcgtt ctagctccct gcttgcccat
     3841 actatatgtt ttaatttata ttttttcttt ccccctggcc ttaaccgaat tttttcccat
     3901 cgcgatctaa ttctcccccg cttaatactg acgctctcgc acccatctct ctccttctag
     3961 cctccgctag tcaaaatttt tggcgtactc accagtcgcc gcccctcgcc tcttgccgtg
     4021 cgcgcttcag caagccgagt cctgcgtcga gagagctcct ctggtttccc tttcgctttc
     4081 aagtccctgt tcgggcgcca ctgctagaga ttttccacac tgactaaaag ggtctgaggg
     4141 atctctagtt accagagtca cacaacagac gggcacacac tacttgaagc actcaaggca
     4201 agctttattg aggcttaagc agtgggttcc ctagttagcc agagagctcc caggctcaga
     4261 tctggtctaa ccagagagac ccagtacagg caaaaagcag ctgcttatat gcaggatctg
     4321 agggctcgcc actccccagt cccgcccagg ccacgcctcc catggtaata gcgatgacta
     4381 atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc
     4441 aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt
     4501 gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag
     4561 tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt
     4621 tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgcggaac tccatatatg
     4681 ggctatgaac taatgacccc gtaattgatt actattaata actagtcaat aatcaatgtc
     4741 gacccaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4801 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4861 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     4921 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     4981 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     5041 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     5101 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     5161 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     5221 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5281 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5341 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5401 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     5461 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     5521 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     5581 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     5641 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     5701 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     5761 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5821 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5881 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5941 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     6001 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     6061 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     6121 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6181 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6241 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6301 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6361 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6421 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6481 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6541 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     6601 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     6661 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     6721 cattaatgca gcggatccag acatgataag atacattgat gagtttggac aaaccacaac
     6781 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     6841 aaccattata agctgcaata aacaagttag ctttttgcaa aagcctaggc ctccaaaaaa
     6901 gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcggcct ctgcataaat
     6961 aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg
     7021 gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgcg gatccgctgg
     7081 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     7141 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     7201 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctt
     7261 gggctgcagg tcgagggatc tccataagag aagagggaca gctatgactg ggagtagtca
     7321 ggagaggagg aaaaatctgg ctagtaaaac atgtaaggaa aattttaggg atgttaaaga
     7381 aaaaaataac acaaaacaaa atat
//
