LOCUS       sequence-488193-        8337 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_173, derived from human fusion protein SAFB2_SAFB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8337)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8337)
  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_173
FEATURES             Location/Qualifiers
     source          1..8337
                     /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(1870..2580,2581..2583,2584..2586))
                     /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(1912..1933)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2261..2280
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2520..2541
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2583..2592
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2668..2871)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2697..2717)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2872..3175
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3432..3476)
                     /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    3661..3894
                     /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    4254..4371
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4863..4988
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5035..5292
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5294..5673
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5704..5808
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5809..5877,5878..6669)
                     /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(6027..6046)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6840..7428
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7329..7348
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7582..7599
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7667..7801
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7691..7710)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7745..7764
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7807..8002)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7821..7956
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    8061..8082
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8097..8102,8103..8120,8121..8127)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8135..8151
                     /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     8140..8162
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8159..8175
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8159..8175
                     /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 gcgcgtcgtc gaactcaaac cggagaccca
      601 caagttacga ccagaacctg aacccacgcg ccccttttcg tccttgataa ttggcttaat
      661 gtctttttcc tctttaaagc tactcatttt ttggtcagca ccttcgctag tgctggactc
      721 cttcggggcg ggtggaacct cattacaggc gtcgaagtcg aatttacgtc catcctcctt
      781 ggaatcgcgg gcttctgggc tcggcgcctc tgtgggggct tcagccagct cctcagaact
      841 cgcttcagct aattcagatg ccgcactact ttgttctacg gcgggctcca gaccgactgg
      901 ttcacagtca gtacgttctc catctcctgg ctgctctgct ggttcacgct tgacgacagc
      961 cagcaggctg tctgccttgc tgctttgggc gtgggccgta ctttccgaag caaggtccag
     1021 gtccccttcc tcagggtgcg cggagtcgaa caggtcctct tcttcggcca gcttacggtc
     1081 aggaccaaca gagcttgtat cttgtgcgaa gggttgctct aattccgaac tttcttcttt
     1141 cacaggctcc gatttacacg tctcaccaag gatgtcaagg atcttctcgt tctctggctc
     1201 caataatgat tcttcaatgt cgggagtaac tttgaagttt aagctggagg tttccaatgt
     1261 attcttaaac ccctccccat ccacggcgtg ctctggcggt tgggcaggta attgacgcaa
     1321 ttgcgccgca acgtactcct tggagtcaca gaaggaatca agcaagccgt cagtcccgtc
     1381 ctcgccaaag tcgggagcac tggagttagc gacctccgtc tcgtctaata cggacatgtc
     1441 catcataccc atattttgta aattctcaag cgaagcctcc atatcttctt gcccatcgcg
     1501 cgagtcgtcc tccagtccgt tgtcctccgt cccctcctct tccattttca atcccttcac
     1561 gcaacgcttg gcagactttt tagaagtggc ctccaactca atcccgattt cgtcggggtc
     1621 ttgtccctct tctttcactg ctttcttcag acgttccatc agtacgcttt tgttcccccc
     1681 cgtatctaag ttacgtttct ttaactctgc gcgtaaatcg atcacgcgaa gctcactcaa
     1741 acgacgtgtc ccagtctctg cgaccccagg acccaagcta gccgtccctg gtccgctgtc
     1801 tcctgaccct ggaagcgttt cggccatccg ggaagcggcc gcacttcccc cggagcctcc
     1861 gcttccgccc ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc
     1921 cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagcttgg actgggtgct
     1981 caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta
     2041 gtggtcggcg agctgcacgc tgccgtcctc gatgttgtgg cggatcttga agttcacctt
     2101 gatgccgttc ttctgcttgt cggccatgat atagacgttg tggctgttgt agttgtactc
     2161 cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg
     2221 gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc
     2281 cttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact tgaagaagtc
     2341 gtgctgcttc atgtggtcgg ggtagcggct gaagcactgc acgccgtagg tcagggtggt
     2401 cacgagggtg ggccagggca cgggcagctt gccggtggtg cagatgaact tcagggtcag
     2461 cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt ggccgtttac
     2521 gtcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgct
     2581 caccatggtg gcgaccggtg gatcccgggc ccgcggtacc gtcgactgca gaattcggtt
     2641 cactaaacga gcccggttca ctaaacgagc tctgcttata tagacctccc accgtacacg
     2701 cctaccgccc atttgcgtca atggggcgga gttgttacga cattttggaa agtcccgttg
     2761 attttggtgc caaaacaaac tcccattgac gtcaatgggg tggagacttg gaaatccccg
     2821 tgagtcaaac cgctatccac gcccattgat gtactgccaa aaccgcatca ccatggtaat
     2881 agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg tcatgtactg
     2941 ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg tacttggcat
     3001 atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca cccattgacg
     3061 tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg acgtcaatgg
     3121 gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg taacgcggaa
     3181 ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactattaat aactagcacg
     3241 cgttcgaagc ttgagctcgg tcccctcggg gttgggaggt gggtctgaaa cgataatggt
     3301 gaatatccct gcctaactct attcactata gaaagtacag caaaaactat tcttaaacct
     3361 accaagcctc ctactatcat tatgaataat tttatatacc acagccaatt tgttatgtta
     3421 aaccaattcc acaaacttgc ccatttatct aattccaata attcttgttc attcttttct
     3481 tgctggtttt gcgattcttc aattaaggag tgtattaagc ttgtgtaatt gttaatttct
     3541 ctgtcccact ccatccaggt cgtgtgattc caaatctgtt ccagagattt attactccaa
     3601 ctagcattcc aaggcacagc agtggtgcaa atgagttttc cagagcaacc ccaaatcccc
     3661 aggagctgtt gatcctttag gtatctttcc acagccagga ttcttgcctg gagctgcttg
     3721 atgccccaga ctgtgagttg caacagatgc tgttgcgcct caatagccct cagcaaattg
     3781 ttctgctgct gcactatacc agacaataat tgtctggcct gtaccgtcag cgtcattgac
     3841 gctgcgccca tagtgcttcc tgctgctccc aagaacccaa ggaacaaagc tcctattccc
     3901 actgctcttt tttctctctg caccactctt ctctttgcct tggtgggtgc tactcctaat
     3961 ggttcaattt ttactacttt atatttatat aattcacttc tccaattgtc cctcatatct
     4021 cctccttatg gtgttttact aaacttttcc atgttctaat cctcatcctg tctacttgcc
     4081 acacaatcat cacctgccat ctgttttcca taatccctaa tgatctttgc ttttcttctt
     4141 ggcactactt ttatgtcact attatcttgt attactactg ccccttcacc tttccagagg
     4201 agctttgctg gtcctttcca aagtggattt ctgctgtccc tgtaataaac ccgaaaattt
     4261 tgaatttttg taatttgttt ttgtaattct ttagtttgta tgtctgttgc tattatgtct
     4321 actattcttt cccctgcact gtacccccca atcccccctt ttcttttaaa attgtggatg
     4381 aatactgcca tttgtactgc tgtcttaaga tgttcagcct gatctcttac ctgtcctata
     4441 attttcttta attctttatt catagattct actactcctt gactttgggg attgtaggga
     4501 atttccaggt ctgaagatcc tgctgcttgc tgtgcttttt tcttactttt gttttgctct
     4561 tcctctatct tgtctaaagc ttccttggtg tcttttatct ctatcctttg atgcacacaa
     4621 tagagggttg ctactgtatt atataatgat ctaagttctt ctgatcctgt ctgaagggat
     4681 ggttgtagct gtcccagtat ttgtctacag ccttctgatg tttctaacag gccagtatta
     4741 actgcgaatc gttctagctc cctgcttgcc catactatat gttttaattt atattttttc
     4801 tttccccctg gccttaaccg aattttttcc catcgcgatc taattctccc ccgcttaata
     4861 ctgacgctct cgcacccatc tctctccttc tagcctccgc tagtcaaaat ttttggcgta
     4921 ctcaccagtc gccgcccctc gcctcttgcc gtgcgcgctt cagcaagccg agtcctgcgt
     4981 cgagagagct cctctggttt ccctttcgct ttcaagtccc tgttcgggcg ccactgctag
     5041 agattttcca cactgactaa aagggtctga gggatctcta gttaccagag tcacacaaca
     5101 gacgggcaca cactacttga agcactcaag gcaagcttta ttgaggctta agcagtgggt
     5161 tccctagtta gccagagagc tcccaggctc agatctggtc taaccagaga gacccagtac
     5221 aggcaaaaag cagctgctta tatgcaggat ctgagggctc gccactcccc agtcccgccc
     5281 aggccacgcc tcccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa
     5341 agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt
     5401 caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc
     5461 gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat
     5521 acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta
     5581 ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg
     5641 attactatta ataactagtc aataatcaat gtcgacccag gtggcacttt tcggggaaat
     5701 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     5761 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     5821 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     5881 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     5941 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     6001 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     6061 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     6121 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     6181 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     6241 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     6301 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     6361 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     6421 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     6481 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     6541 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     6601 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     6661 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     6721 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     6781 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     6841 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     6901 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     6961 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     7021 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     7081 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     7141 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     7201 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     7261 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     7321 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     7381 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     7441 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     7501 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     7561 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     7621 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagcggatc cagacatgat
     7681 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     7741 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     7801 tagctttttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc
     7861 agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg
     7921 agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat
     7981 ggttgctgac taattgagat gcggatccgc tggcacgaca ggtttcccga ctggaaagcg
     8041 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     8101 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     8161 ggaaacagct atgaccatga ttacgccaag cttgggctgc aggtcgaggg atctccataa
     8221 gagaagaggg acagctatga ctgggagtag tcaggagagg aggaaaaatc tggctagtaa
     8281 aacatgtaag gaaaatttta gggatgttaa agaaaaaaat aacacaaaac aaaatat
//
