LOCUS       sequence-488186-        8277 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_166, derived from human fusion protein RCSD1_ABL2; 
            ZC3HAV1_ABL2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8277)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8277)
  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_166
FEATURES             Location/Qualifiers
     source          1..8277
                     /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(1810..2520,2521..2523,2524..2526))
                     /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(1852..1873)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2201..2220
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2460..2481
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2523..2532
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2608..2811)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2637..2657)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2812..3115
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3372..3416)
                     /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    3601..3834
                     /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    4194..4311
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4803..4928
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4975..5232
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5234..5613
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5644..5748
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5749..5817,5818..6609)
                     /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(5967..5986)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6780..7368
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7269..7288
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7522..7539
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7607..7741
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7631..7650)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7685..7704
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7747..7942)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7761..7896
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    8001..8022
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8037..8042,8043..8060,8061..8067)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8075..8091
                     /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     8080..8102
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8099..8115
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8099..8115
                     /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 tttctcagcg gcctgtttgg tttttctcag
      601 agccacttta gtacctgctg tgccattggc cattttggct ggcgagatgg aagatgtggg
      661 cagaggcact tgaggtggag gcatcactgg cctcccagct ttcccactga tgggcactgc
      721 gcccagagct gccttctttc ctccttcctg tgtttctgat gtggactgtc ctgcagttag
      781 ggcagttggc tcttctgtag ggtctgagca gatggacgga tgctgcagta gtctcatcac
      841 tggtggtggg ggtggggcac actttggttt tacccgtcgg ggtcggtcct tgtctccaga
      901 ggatgtgacc tgatgctcag ataagagctt gaatttattc ccctgagagt ctgtgccaat
      961 gagctgcacg tcagctggag tgtgtttcag agtgggtgag ataaggactg gcactttgtg
     1021 gttgtgagtg gttgggagga cgggggcagc cttggctgga gaaggccagc ccggctgctc
     1081 tccatcctct ggaactccag ccatcccaag tcgtgcccca ccattcttct ctttaccctt
     1141 gggggcagct gccactccag ccactcccac ccctggaggg tccttctctg taatggctag
     1201 atccccagag ggtgttctga gaggaagagc tgtggctcct ctgggcaata acttggcttt
     1261 tggtctctcc ctgcttggag cagcactttc ctctgatttt tttggaagca tgtcattggc
     1321 cctgtccaca ttctcttctg gctgagaaga ggtggacact gtcctttcca gctggagttt
     1381 ggacctctgg cagttcctgg gaagggtcat tgccatccta tcctgctctg gaagccccgt
     1441 gagttcgtat ttcttatggg gctgattctc catttctcgg aaggagctgc tgcgtttggg
     1501 gggtgtagga gcatttctct tcttcatgaa ggagctgaag aagcccccct tcctatccct
     1561 ggtgaagcat gtctctttgg catcttccaa gaggctgctg ggtgacttgt ctctttgctt
     1621 tcgaggcagt gctggggatc cactagaggc ctgtgcacct ctgatgaacc ctggtgctaa
     1681 actggaagca gaattttctg tggcatcttg tgccccttca atgttctcct tgttctccac
     1741 ctgtttcttc agtgtccgag tcttggaccg ggaagcggcc gcacttcccc cggagcctcc
     1801 gcttccgccc ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc
     1861 cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagcttgg actgggtgct
     1921 caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta
     1981 gtggtcggcg agctgcacgc tgccgtcctc gatgttgtgg cggatcttga agttcacctt
     2041 gatgccgttc ttctgcttgt cggccatgat atagacgttg tggctgttgt agttgtactc
     2101 cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg
     2161 gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc
     2221 cttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact tgaagaagtc
     2281 gtgctgcttc atgtggtcgg ggtagcggct gaagcactgc acgccgtagg tcagggtggt
     2341 cacgagggtg ggccagggca cgggcagctt gccggtggtg cagatgaact tcagggtcag
     2401 cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt ggccgtttac
     2461 gtcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgct
     2521 caccatggtg gcgaccggtg gatcccgggc ccgcggtacc gtcgactgca gaattcggtt
     2581 cactaaacga gcccggttca ctaaacgagc tctgcttata tagacctccc accgtacacg
     2641 cctaccgccc atttgcgtca atggggcgga gttgttacga cattttggaa agtcccgttg
     2701 attttggtgc caaaacaaac tcccattgac gtcaatgggg tggagacttg gaaatccccg
     2761 tgagtcaaac cgctatccac gcccattgat gtactgccaa aaccgcatca ccatggtaat
     2821 agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg tcatgtactg
     2881 ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg tacttggcat
     2941 atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca cccattgacg
     3001 tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg acgtcaatgg
     3061 gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg taacgcggaa
     3121 ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactattaat aactagcacg
     3181 cgttcgaagc ttgagctcgg tcccctcggg gttgggaggt gggtctgaaa cgataatggt
     3241 gaatatccct gcctaactct attcactata gaaagtacag caaaaactat tcttaaacct
     3301 accaagcctc ctactatcat tatgaataat tttatatacc acagccaatt tgttatgtta
     3361 aaccaattcc acaaacttgc ccatttatct aattccaata attcttgttc attcttttct
     3421 tgctggtttt gcgattcttc aattaaggag tgtattaagc ttgtgtaatt gttaatttct
     3481 ctgtcccact ccatccaggt cgtgtgattc caaatctgtt ccagagattt attactccaa
     3541 ctagcattcc aaggcacagc agtggtgcaa atgagttttc cagagcaacc ccaaatcccc
     3601 aggagctgtt gatcctttag gtatctttcc acagccagga ttcttgcctg gagctgcttg
     3661 atgccccaga ctgtgagttg caacagatgc tgttgcgcct caatagccct cagcaaattg
     3721 ttctgctgct gcactatacc agacaataat tgtctggcct gtaccgtcag cgtcattgac
     3781 gctgcgccca tagtgcttcc tgctgctccc aagaacccaa ggaacaaagc tcctattccc
     3841 actgctcttt tttctctctg caccactctt ctctttgcct tggtgggtgc tactcctaat
     3901 ggttcaattt ttactacttt atatttatat aattcacttc tccaattgtc cctcatatct
     3961 cctccttatg gtgttttact aaacttttcc atgttctaat cctcatcctg tctacttgcc
     4021 acacaatcat cacctgccat ctgttttcca taatccctaa tgatctttgc ttttcttctt
     4081 ggcactactt ttatgtcact attatcttgt attactactg ccccttcacc tttccagagg
     4141 agctttgctg gtcctttcca aagtggattt ctgctgtccc tgtaataaac ccgaaaattt
     4201 tgaatttttg taatttgttt ttgtaattct ttagtttgta tgtctgttgc tattatgtct
     4261 actattcttt cccctgcact gtacccccca atcccccctt ttcttttaaa attgtggatg
     4321 aatactgcca tttgtactgc tgtcttaaga tgttcagcct gatctcttac ctgtcctata
     4381 attttcttta attctttatt catagattct actactcctt gactttgggg attgtaggga
     4441 atttccaggt ctgaagatcc tgctgcttgc tgtgcttttt tcttactttt gttttgctct
     4501 tcctctatct tgtctaaagc ttccttggtg tcttttatct ctatcctttg atgcacacaa
     4561 tagagggttg ctactgtatt atataatgat ctaagttctt ctgatcctgt ctgaagggat
     4621 ggttgtagct gtcccagtat ttgtctacag ccttctgatg tttctaacag gccagtatta
     4681 actgcgaatc gttctagctc cctgcttgcc catactatat gttttaattt atattttttc
     4741 tttccccctg gccttaaccg aattttttcc catcgcgatc taattctccc ccgcttaata
     4801 ctgacgctct cgcacccatc tctctccttc tagcctccgc tagtcaaaat ttttggcgta
     4861 ctcaccagtc gccgcccctc gcctcttgcc gtgcgcgctt cagcaagccg agtcctgcgt
     4921 cgagagagct cctctggttt ccctttcgct ttcaagtccc tgttcgggcg ccactgctag
     4981 agattttcca cactgactaa aagggtctga gggatctcta gttaccagag tcacacaaca
     5041 gacgggcaca cactacttga agcactcaag gcaagcttta ttgaggctta agcagtgggt
     5101 tccctagtta gccagagagc tcccaggctc agatctggtc taaccagaga gacccagtac
     5161 aggcaaaaag cagctgctta tatgcaggat ctgagggctc gccactcccc agtcccgccc
     5221 aggccacgcc tcccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa
     5281 agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt
     5341 caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc
     5401 gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat
     5461 acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta
     5521 ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg
     5581 attactatta ataactagtc aataatcaat gtcgacccag gtggcacttt tcggggaaat
     5641 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     5701 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     5761 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     5821 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     5881 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     5941 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     6001 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     6061 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     6121 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     6181 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     6241 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     6301 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     6361 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     6421 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     6481 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     6541 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     6601 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     6661 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     6721 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     6781 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     6841 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     6901 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     6961 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     7021 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     7081 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     7141 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     7201 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     7261 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     7321 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     7381 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     7441 ttatcccctg cttctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     7501 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     7561 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagcggatc cagacatgat
     7621 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     7681 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     7741 tagctttttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc
     7801 agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg
     7861 agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat
     7921 ggttgctgac taattgagat gcggatccgc tggcacgaca ggtttcccga ctggaaagcg
     7981 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     8041 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     8101 ggaaacagct atgaccatga ttacgccaag cttgggctgc aggtcgaggg atctccataa
     8161 gagaagaggg acagctatga ctgggagtag tcaggagagg aggaaaaatc tggctagtaa
     8221 aacatgtaag gaaaatttta gggatgttaa agaaaaaaat aacacaaaac aaaatat
//
