LOCUS       sequence-490713-        7272 bp    DNA     circular SYN 27-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_032, derived from human fusion protein CBFA2T3_GLIS2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7272)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7272)
  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_032
FEATURES             Location/Qualifiers
     source          1..7272
                     /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(805..1515,1516..1518,1519..1521))
                     /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(847..868)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1196..1215
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1455..1476
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1518..1527
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1603..1806)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1632..1652)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1807..2110
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2367..2411)
                     /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    2596..2829
                     /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    3189..3306
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    3798..3923
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             3970..4227
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4229..4608
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4639..4743
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4744..4812,4813..5604)
                     /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(4962..4981)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5775..6363
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6264..6283
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6517..6534
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6602..6736
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6626..6645)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6680..6699
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6742..6937)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6756..6891
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    6996..7017
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7032..7037,7038..7055,7056..7062)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7070..7086
                     /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     7075..7097
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7094..7110
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7094..7110
                     /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 cggggaactc gccgcagagt taacgccggt
      601 cgataaatct aacaccgtac cgggaagaag gggaaggccg tcgggactgg gacggctgca
      661 tgatgactcg gtgcgttcca ggggtgtttt gtgtccttcc atgccaagag cgcgggttgg
      721 tacagaaccg cgccctttct ctccctcagc tttaccccct gcccgggaag cggccgcact
      781 tcccccggag cctccgcttc cgcccttgta cagctcgtcc atgccgagag tgatcccggc
      841 ggcggtcacg aactccagca ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag
      901 cttggactgg gtgctcaggt agtggttgtc gggcagcagc acggggccgt cgccgatggg
      961 ggtgttctgc tggtagtggt cggcgagctg cacgctgccg tcctcgatgt tgtggcggat
     1021 cttgaagttc accttgatgc cgttcttctg cttgtcggcc atgatataga cgttgtggct
     1081 gttgtagttg tactccagct tgtgccccag gatgttgccg tcctccttga agtcgatgcc
     1141 cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt
     1201 gtagttgccg tcgtccttga agaagatggt gcgctcctgg acgtagcctt cgggcatggc
     1261 ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag cggctgaagc actgcacgcc
     1321 gtaggtcagg gtggtcacga gggtgggcca gggcacgggc agcttgccgg tggtgcagat
     1381 gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa
     1441 cttgtggccg tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc cggtgaacag
     1501 ctcctcgccc ttgctcacca tggtggcgac cggtggatcc cgggcccgcg gtaccgtcga
     1561 ctgcagaatt cggttcacta aacgagcccg gttcactaaa cgagctctgc ttatatagac
     1621 ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt tacgacattt
     1681 tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggggtggag
     1741 acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact gccaaaaccg
     1801 catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca
     1861 taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg
     1921 gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata
     1981 ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat
     2041 tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag
     2101 ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta
     2161 ttaataacta gcacgcgttc gaagcttgag ctcggtcccc tcggggttgg gaggtgggtc
     2221 tgaaacgata atggtgaata tccctgccta actctattca ctatagaaag tacagcaaaa
     2281 actattctta aacctaccaa gcctcctact atcattatga ataattttat ataccacagc
     2341 caatttgtta tgttaaacca attccacaaa cttgcccatt tatctaattc caataattct
     2401 tgttcattct tttcttgctg gttttgcgat tcttcaatta aggagtgtat taagcttgtg
     2461 taattgttaa tttctctgtc ccactccatc caggtcgtgt gattccaaat ctgttccaga
     2521 gatttattac tccaactagc attccaaggc acagcagtgg tgcaaatgag ttttccagag
     2581 caaccccaaa tccccaggag ctgttgatcc tttaggtatc tttccacagc caggattctt
     2641 gcctggagct gcttgatgcc ccagactgtg agttgcaaca gatgctgttg cgcctcaata
     2701 gccctcagca aattgttctg ctgctgcact ataccagaca ataattgtct ggcctgtacc
     2761 gtcagcgtca ttgacgctgc gcccatagtg cttcctgctg ctcccaagaa cccaaggaac
     2821 aaagctccta ttcccactgc tcttttttct ctctgcacca ctcttctctt tgccttggtg
     2881 ggtgctactc ctaatggttc aatttttact actttatatt tatataattc acttctccaa
     2941 ttgtccctca tatctcctcc ttatggtgtt ttactaaact tttccatgtt ctaatcctca
     3001 tcctgtctac ttgccacaca atcatcacct gccatctgtt ttccataatc cctaatgatc
     3061 tttgcttttc ttcttggcac tacttttatg tcactattat cttgtattac tactgcccct
     3121 tcacctttcc agaggagctt tgctggtcct ttccaaagtg gatttctgct gtccctgtaa
     3181 taaacccgaa aattttgaat ttttgtaatt tgtttttgta attctttagt ttgtatgtct
     3241 gttgctatta tgtctactat tctttcccct gcactgtacc ccccaatccc cccttttctt
     3301 ttaaaattgt ggatgaatac tgccatttgt actgctgtct taagatgttc agcctgatct
     3361 cttacctgtc ctataatttt ctttaattct ttattcatag attctactac tccttgactt
     3421 tggggattgt agggaatttc caggtctgaa gatcctgctg cttgctgtgc ttttttctta
     3481 cttttgtttt gctcttcctc tatcttgtct aaagcttcct tggtgtcttt tatctctatc
     3541 ctttgatgca cacaatagag ggttgctact gtattatata atgatctaag ttcttctgat
     3601 cctgtctgaa gggatggttg tagctgtccc agtatttgtc tacagccttc tgatgtttct
     3661 aacaggccag tattaactgc gaatcgttct agctccctgc ttgcccatac tatatgtttt
     3721 aatttatatt ttttctttcc ccctggcctt aaccgaattt tttcccatcg cgatctaatt
     3781 ctcccccgct taatactgac gctctcgcac ccatctctct ccttctagcc tccgctagtc
     3841 aaaatttttg gcgtactcac cagtcgccgc ccctcgcctc ttgccgtgcg cgcttcagca
     3901 agccgagtcc tgcgtcgaga gagctcctct ggtttccctt tcgctttcaa gtccctgttc
     3961 gggcgccact gctagagatt ttccacactg actaaaaggg tctgagggat ctctagttac
     4021 cagagtcaca caacagacgg gcacacacta cttgaagcac tcaaggcaag ctttattgag
     4081 gcttaagcag tgggttccct agttagccag agagctccca ggctcagatc tggtctaacc
     4141 agagagaccc agtacaggca aaaagcagct gcttatatgc aggatctgag ggctcgccac
     4201 tccccagtcc cgcccaggcc acgcctccca tggtaatagc gatgactaat acgtagatgt
     4261 actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt
     4321 taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca
     4381 agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg
     4441 ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc
     4501 caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta
     4561 atgaccccgt aattgattac tattaataac tagtcaataa tcaatgtcga cccaggtggc
     4621 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     4681 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4741 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     4801 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     4861 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     4921 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     4981 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5041 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5101 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5161 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5221 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5281 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5341 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5401 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     5461 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     5521 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     5581 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     5641 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5701 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5761 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5821 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5881 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     5941 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6001 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6061 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6121 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6181 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6241 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6301 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6361 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6421 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     6481 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     6541 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     6601 ggatccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     6661 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     6721 ctgcaataaa caagttagct ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta
     6781 cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag
     6841 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
     6901 taggggcggg actatggttg ctgactaatt gagatgcgga tccgctggca cgacaggttt
     6961 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     7021 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     7081 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgg gctgcaggtc
     7141 gagggatctc cataagagaa gagggacagc tatgactggg agtagtcagg agaggaggaa
     7201 aaatctggct agtaaaacat gtaaggaaaa ttttagggat gttaaagaaa aaaataacac
     7261 aaaacaaaat at
//
