LOCUS       sequence-487975-        7878 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_065, derived from human fusion protein ETV6_RUNX1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7878)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7878)
  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_065
FEATURES             Location/Qualifiers
     source          1..7878
                     /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(1411..2121,2122..2124,2125..2127))
                     /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(1453..1474)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1802..1821
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2061..2082
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2124..2133
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2209..2412)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2238..2258)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2413..2716
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2973..3017)
                     /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    3202..3435
                     /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    3795..3912
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4404..4529
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4576..4833
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4835..5214
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5245..5349
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5350..5418,5419..6210)
                     /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(5568..5587)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6381..6969
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6870..6889
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7123..7140
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7208..7342
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7232..7251)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7286..7305
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7348..7543)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7362..7497
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7602..7623
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7638..7643,7644..7661,7662..7668)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7676..7692
                     /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     7681..7703
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7700..7716
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7700..7716
                     /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 ggggtgatca gcaagcactt ccaccatgct
      601 gcggtccccg ctgcgcagct tccccgccag agcagcacca gcatcaggcg ctcccaacgg
      661 taaggcctcg gacatcttcc caggtgataa cgccgtacta ggcggcgtaa agcggcggct
      721 ggttgatgcg tcgtgcacgt cgcgtgacgg attcattccc agaatgcatt ccgcgatgcg
      781 accgatcggc atagcatgtt cctcgggtgg agagaccgac accatgatgt gattcatgta
      841 agccagatct tcgcgatgag acaagttgat cggcttgcct tcgcgatgca ggccgtcctc
      901 ggacaagcga gattgcttga aatctacaga atggcgggga ttcaaaatca gcgggtgcat
      961 aatgggtgac ggcattaact ggataacacg agtagactct tggcgtggac ttgatggctt
     1021 tgggtgactc tcgcttgacg ctgggcagtg gttattctcc ataggcgaca cagacagagg
     1081 atagctttcc tggtggttat tttcctggtg ggggcgtggt ccttgggcgc gctcagctgg
     1141 agaaagacgg cgaatcatat tatcaagtgg actgcgcaaa gggcgttgtt cgggatcggg
     1201 gctggggcga tgattcgtag tgattgggct acgagagcgg tgaagaagct caatcgtggg
     1261 cggattgtgg tgtacattgt caaccgatgg gcgcggcgtg cgctggacac aattgtcctc
     1321 ctcgtgattt tgatgtaaaa tgacctctgg ctgagtgtgg atggaattcc gggaagcggc
     1381 cgcacttccc ccggagcctc cgcttccgcc cttgtacagc tcgtccatgc cgagagtgat
     1441 cccggcggcg gtcacgaact ccagcaggac catgtgatcg cgcttctcgt tggggtcttt
     1501 gctcagcttg gactgggtgc tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc
     1561 gatgggggtg ttctgctggt agtggtcggc gagctgcacg ctgccgtcct cgatgttgtg
     1621 gcggatcttg aagttcacct tgatgccgtt cttctgcttg tcggccatga tatagacgtt
     1681 gtggctgttg tagttgtact ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc
     1741 gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg
     1801 ggtcttgtag ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt agccttcggg
     1861 catggcggac ttgaagaagt cgtgctgctt catgtggtcg gggtagcggc tgaagcactg
     1921 cacgccgtag gtcagggtgg tcacgagggt gggccagggc acgggcagct tgccggtggt
     1981 gcagatgaac ttcagggtca gcttgccgta ggtggcatcg ccctcgccct cgccggacac
     2041 gctgaacttg tggccgttta cgtcgccgtc cagctcgacc aggatgggca ccaccccggt
     2101 gaacagctcc tcgcccttgc tcaccatggt ggcgaccggt ggatcccggg cccgcggtac
     2161 cgtcgactgc agaattcggt tcactaaacg agcccggttc actaaacgag ctctgcttat
     2221 atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg agttgttacg
     2281 acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga cgtcaatggg
     2341 gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga tgtactgcca
     2401 aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa
     2461 gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc
     2521 aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg
     2581 taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata
     2641 cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac
     2701 cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga
     2761 ttactattaa taactagcac gcgttcgaag cttgagctcg gtcccctcgg ggttgggagg
     2821 tgggtctgaa acgataatgg tgaatatccc tgcctaactc tattcactat agaaagtaca
     2881 gcaaaaacta ttcttaaacc taccaagcct cctactatca ttatgaataa ttttatatac
     2941 cacagccaat ttgttatgtt aaaccaattc cacaaacttg cccatttatc taattccaat
     3001 aattcttgtt cattcttttc ttgctggttt tgcgattctt caattaagga gtgtattaag
     3061 cttgtgtaat tgttaatttc tctgtcccac tccatccagg tcgtgtgatt ccaaatctgt
     3121 tccagagatt tattactcca actagcattc caaggcacag cagtggtgca aatgagtttt
     3181 ccagagcaac cccaaatccc caggagctgt tgatccttta ggtatctttc cacagccagg
     3241 attcttgcct ggagctgctt gatgccccag actgtgagtt gcaacagatg ctgttgcgcc
     3301 tcaatagccc tcagcaaatt gttctgctgc tgcactatac cagacaataa ttgtctggcc
     3361 tgtaccgtca gcgtcattga cgctgcgccc atagtgcttc ctgctgctcc caagaaccca
     3421 aggaacaaag ctcctattcc cactgctctt ttttctctct gcaccactct tctctttgcc
     3481 ttggtgggtg ctactcctaa tggttcaatt tttactactt tatatttata taattcactt
     3541 ctccaattgt ccctcatatc tcctccttat ggtgttttac taaacttttc catgttctaa
     3601 tcctcatcct gtctacttgc cacacaatca tcacctgcca tctgttttcc ataatcccta
     3661 atgatctttg cttttcttct tggcactact tttatgtcac tattatcttg tattactact
     3721 gccccttcac ctttccagag gagctttgct ggtcctttcc aaagtggatt tctgctgtcc
     3781 ctgtaataaa cccgaaaatt ttgaattttt gtaatttgtt tttgtaattc tttagtttgt
     3841 atgtctgttg ctattatgtc tactattctt tcccctgcac tgtacccccc aatcccccct
     3901 tttcttttaa aattgtggat gaatactgcc atttgtactg ctgtcttaag atgttcagcc
     3961 tgatctctta cctgtcctat aattttcttt aattctttat tcatagattc tactactcct
     4021 tgactttggg gattgtaggg aatttccagg tctgaagatc ctgctgcttg ctgtgctttt
     4081 ttcttacttt tgttttgctc ttcctctatc ttgtctaaag cttccttggt gtcttttatc
     4141 tctatccttt gatgcacaca atagagggtt gctactgtat tatataatga tctaagttct
     4201 tctgatcctg tctgaaggga tggttgtagc tgtcccagta tttgtctaca gccttctgat
     4261 gtttctaaca ggccagtatt aactgcgaat cgttctagct ccctgcttgc ccatactata
     4321 tgttttaatt tatatttttt ctttccccct ggccttaacc gaattttttc ccatcgcgat
     4381 ctaattctcc cccgcttaat actgacgctc tcgcacccat ctctctcctt ctagcctccg
     4441 ctagtcaaaa tttttggcgt actcaccagt cgccgcccct cgcctcttgc cgtgcgcgct
     4501 tcagcaagcc gagtcctgcg tcgagagagc tcctctggtt tccctttcgc tttcaagtcc
     4561 ctgttcgggc gccactgcta gagattttcc acactgacta aaagggtctg agggatctct
     4621 agttaccaga gtcacacaac agacgggcac acactacttg aagcactcaa ggcaagcttt
     4681 attgaggctt aagcagtggg ttccctagtt agccagagag ctcccaggct cagatctggt
     4741 ctaaccagag agacccagta caggcaaaaa gcagctgctt atatgcagga tctgagggct
     4801 cgccactccc cagtcccgcc caggccacgc ctcccatggt aatagcgatg actaatacgt
     4861 agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa tgccaggcgg
     4921 gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac acttgatgta
     4981 ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta
     5041 ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg
     5101 gtcagccagg cgggccattt accgtaagtt atgtaacgcg gaactccata tatgggctat
     5161 gaactaatga ccccgtaatt gattactatt aataactagt caataatcaa tgtcgaccca
     5221 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     5281 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     5341 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     5401 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     5461 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     5521 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     5581 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     5641 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     5701 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     5761 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     5821 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     5881 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     5941 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     6001 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     6061 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     6121 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     6181 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     6241 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6301 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6361 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     6421 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     6481 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     6541 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6601 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6661 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6721 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6781 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6841 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6901 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6961 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     7021 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     7081 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     7141 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     7201 tgcagcggat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     7261 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     7321 tataagctgc aataaacaag ttagcttttt gcaaaagcct aggcctccaa aaaagcctcc
     7381 tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa
     7441 aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg
     7501 cggagttagg ggcgggacta tggttgctga ctaattgaga tgcggatccg ctggcacgac
     7561 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     7621 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     7681 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcttgggctg
     7741 caggtcgagg gatctccata agagaagagg gacagctatg actgggagta gtcaggagag
     7801 gaggaaaaat ctggctagta aaacatgtaa ggaaaatttt agggatgtta aagaaaaaaa
     7861 taacacaaaa caaaatat
//
