LOCUS       sequence-488227-        7959 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_212, derived from human fusion protein ZC3HAV1_ABL2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7959)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7959)
  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_212
FEATURES             Location/Qualifiers
     source          1..7959
                     /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(1492..2202,2203..2205,2206..2208))
                     /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(1534..1555)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1883..1902
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2142..2163
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2205..2214
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2290..2493)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2319..2339)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2494..2797
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3054..3098)
                     /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    3283..3516
                     /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    3876..3993
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4485..4610
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4657..4914
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4916..5295
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5326..5430
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5431..5499,5500..6291)
                     /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(5649..5668)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6462..7050
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6951..6970
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7204..7221
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7289..7423
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7313..7332)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7367..7386
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7429..7624)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7443..7578
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7683..7704
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7719..7724,7725..7742,7743..7749)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7757..7773
                     /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     7762..7784
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7781..7797
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7781..7797
                     /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 atccacccta gaagatgtgg tacttgtgac
      601 atcagataaa gaatcgttaa caagtgctac tcttgggtca gcatcatctg cgattctgcc
      661 agtggcctgg acatctcggg aagcaggtcc agcatcctga atcctaggtg atgatatttc
      721 tctgtgaccg ctgctagtgc ttttgtaatt taaggatctg gtagaagtta tatctgtggc
      781 cactccatca gcattattat taaaaagagg gccaggctgg atgtcctgag ttccactttt
      841 gccattgatg atcctgtagt ctgaggaaag caagcctgtg ccctttctgg tggtcacagc
      901 ttcaggtgtt tgcagagagc caagagaaga gcgggcggca ggtagcgtgg gagaaaacac
      961 agtctttctc ctggcgcctt ggtcattcgt ccaggatgtg aggctcttcc agttgggggc
     1021 tgatgttgaa ttggaagcaa ggtaagtgct gcctggattt ccatgcaaga ggtcctcttg
     1081 actgccgttc tctaaaaacc tctggcttgt cccggcctga cttgttcctc caagatcagt
     1141 agccttggac gagcctgagg gaggccgagc gcgatcctga ctccccagat acgtgaactt
     1201 gcgggtgaga tcgtccacag gcgcgtcctc cagggaagcc ctgtggctga tctgatctgg
     1261 actaggtgtg caggacctct cagcagacgc tgaagcagac gcaagaaatt cttggctgcc
     1321 ctgaaagaac cgatctctac tcttgcttct agccctatat gccatgtttc tacgatgtga
     1381 agaaggagct ctgggccctg ggggattctt ctgcatgtgc ttgctgttgc agatgtcctg
     1441 gatgttctgc cgggaagcgg ccgcacttcc cccggagcct ccgcttccgc ccttgtacag
     1501 ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga ccatgtgatc
     1561 gcgcttctcg ttggggtctt tgctcagctt ggactgggtg ctcaggtagt ggttgtcggg
     1621 cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg cgagctgcac
     1681 gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc ttgatgccgt tcttctgctt
     1741 gtcggccatg atatagacgt tgtggctgtt gtagttgtac tccagcttgt gccccaggat
     1801 gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca gggtgtcgcc
     1861 ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga agatggtgcg
     1921 ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc
     1981 ggggtagcgg ctgaagcact gcacgccgta ggtcagggtg gtcacgaggg tgggccaggg
     2041 cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt aggtggcatc
     2101 gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt ccagctcgac
     2161 caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatgg tggcgaccgg
     2221 tggatcccgg gcccgcggta ccgtcgactg cagaattcgg ttcactaaac gagcccggtt
     2281 cactaaacga gctctgctta tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt
     2341 caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt gccaaaacaa
     2401 actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa accgctatcc
     2461 acgcccattg atgtactgcc aaaaccgcat caccatggta atagcgatga ctaatacgta
     2521 gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg
     2581 ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac
     2641 tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat
     2701 tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg
     2761 tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat atgggctatg
     2821 aactaatgac cccgtaattg attactatta ataactagca cgcgttcgaa gcttgagctc
     2881 ggtcccctcg gggttgggag gtgggtctga aacgataatg gtgaatatcc ctgcctaact
     2941 ctattcacta tagaaagtac agcaaaaact attcttaaac ctaccaagcc tcctactatc
     3001 attatgaata attttatata ccacagccaa tttgttatgt taaaccaatt ccacaaactt
     3061 gcccatttat ctaattccaa taattcttgt tcattctttt cttgctggtt ttgcgattct
     3121 tcaattaagg agtgtattaa gcttgtgtaa ttgttaattt ctctgtccca ctccatccag
     3181 gtcgtgtgat tccaaatctg ttccagagat ttattactcc aactagcatt ccaaggcaca
     3241 gcagtggtgc aaatgagttt tccagagcaa ccccaaatcc ccaggagctg ttgatccttt
     3301 aggtatcttt ccacagccag gattcttgcc tggagctgct tgatgcccca gactgtgagt
     3361 tgcaacagat gctgttgcgc ctcaatagcc ctcagcaaat tgttctgctg ctgcactata
     3421 ccagacaata attgtctggc ctgtaccgtc agcgtcattg acgctgcgcc catagtgctt
     3481 cctgctgctc ccaagaaccc aaggaacaaa gctcctattc ccactgctct tttttctctc
     3541 tgcaccactc ttctctttgc cttggtgggt gctactccta atggttcaat ttttactact
     3601 ttatatttat ataattcact tctccaattg tccctcatat ctcctcctta tggtgtttta
     3661 ctaaactttt ccatgttcta atcctcatcc tgtctacttg ccacacaatc atcacctgcc
     3721 atctgttttc cataatccct aatgatcttt gcttttcttc ttggcactac ttttatgtca
     3781 ctattatctt gtattactac tgccccttca cctttccaga ggagctttgc tggtcctttc
     3841 caaagtggat ttctgctgtc cctgtaataa acccgaaaat tttgaatttt tgtaatttgt
     3901 ttttgtaatt ctttagtttg tatgtctgtt gctattatgt ctactattct ttcccctgca
     3961 ctgtaccccc caatcccccc ttttctttta aaattgtgga tgaatactgc catttgtact
     4021 gctgtcttaa gatgttcagc ctgatctctt acctgtccta taattttctt taattcttta
     4081 ttcatagatt ctactactcc ttgactttgg ggattgtagg gaatttccag gtctgaagat
     4141 cctgctgctt gctgtgcttt tttcttactt ttgttttgct cttcctctat cttgtctaaa
     4201 gcttccttgg tgtcttttat ctctatcctt tgatgcacac aatagagggt tgctactgta
     4261 ttatataatg atctaagttc ttctgatcct gtctgaaggg atggttgtag ctgtcccagt
     4321 atttgtctac agccttctga tgtttctaac aggccagtat taactgcgaa tcgttctagc
     4381 tccctgcttg cccatactat atgttttaat ttatattttt tctttccccc tggccttaac
     4441 cgaatttttt cccatcgcga tctaattctc ccccgcttaa tactgacgct ctcgcaccca
     4501 tctctctcct tctagcctcc gctagtcaaa atttttggcg tactcaccag tcgccgcccc
     4561 tcgcctcttg ccgtgcgcgc ttcagcaagc cgagtcctgc gtcgagagag ctcctctggt
     4621 ttccctttcg ctttcaagtc cctgttcggg cgccactgct agagattttc cacactgact
     4681 aaaagggtct gagggatctc tagttaccag agtcacacaa cagacgggca cacactactt
     4741 gaagcactca aggcaagctt tattgaggct taagcagtgg gttccctagt tagccagaga
     4801 gctcccaggc tcagatctgg tctaaccaga gagacccagt acaggcaaaa agcagctgct
     4861 tatatgcagg atctgagggc tcgccactcc ccagtcccgc ccaggccacg cctcccatgg
     4921 taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt
     4981 actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg
     5041 gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt
     5101 gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca
     5161 atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc
     5221 ggaactccat atatgggcta tgaactaatg accccgtaat tgattactat taataactag
     5281 tcaataatca atgtcgaccc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5341 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5401 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     5461 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5521 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5581 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5641 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5701 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5761 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5821 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5881 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5941 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     6001 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     6061 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     6121 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     6181 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6241 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     6301 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6361 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6421 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6481 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6541 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6601 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6661 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6721 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6781 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6841 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6901 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6961 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     7021 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     7081 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     7141 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     7201 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     7261 gcgcgttggc cgattcatta atgcagcgga tccagacatg ataagataca ttgatgagtt
     7321 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     7381 tattgcttta tttgtaacca ttataagctg caataaacaa gttagctttt tgcaaaagcc
     7441 taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc
     7501 ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg
     7561 cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag
     7621 atgcggatcc gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     7681 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     7741 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     7801 gattacgcca agcttgggct gcaggtcgag ggatctccat aagagaagag ggacagctat
     7861 gactgggagt agtcaggaga ggaggaaaaa tctggctagt aaaacatgta aggaaaattt
     7921 tagggatgtt aaagaaaaaa ataacacaaa acaaaatat
//
