LOCUS       sequence-488203-        7833 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_184, derived from human fusion protein SSBP2_JAK2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7833)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7833)
  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_184
FEATURES             Location/Qualifiers
     source          1..7833
                     /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(1366..2076,2077..2079,2080..2082))
                     /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(1408..1429)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1757..1776
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2016..2037
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2079..2088
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2164..2367)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2193..2213)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2368..2671
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2928..2972)
                     /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    3157..3390
                     /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    3750..3867
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4359..4484
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4531..4788
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4790..5169
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5200..5304
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5305..5373,5374..6165)
                     /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(5523..5542)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6336..6924
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6825..6844
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7078..7095
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7163..7297
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7187..7206)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7241..7260
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7303..7498)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7317..7452
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7557..7578
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7593..7598,7599..7616,7617..7623)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7631..7647
                     /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     7636..7658
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7655..7671
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7655..7671
                     /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 cccaagggac tcattcttgc taatactatc
      601 catatctccg gagcccaatg atccattcat gtggtggctt tccattccgc ctaagccacc
      661 cataggaccg tctgaccctg ggcccattgg gaagttcggg cggttcggac caggaggaac
      721 ggcattcata agggtataca tgttgtcgcc cgaatttgta gaatcagccg ggctaggcat
      781 aattggcgta cccggtggcc cgccaccacc gggtgggccc acatagttac ctggggatgc
      841 actactgtat gggatagagt tcgcgttagt agggttgggc catgggcgtc ccccaccggg
      901 gcccatgttc attccaggca tgccgggtcc tcccaaggcg ttcagtggcg ggcgcattgc
      961 tccgccgtaa ttctgtaaag acaaccaggg gtcagactga ggcccaagcg gcaccatacc
     1021 gcggggcgga gtcatacgtt gcatcggccc acccatattt ggatgcccct gctgacgggt
     1081 tgggtccatg cctgatggca gcagtggctg tgatccaggc acgccgccaa gagcttggtt
     1141 tggaatgcgt aacggcgggc gaggaccacc cggataacgc ggactcatga agggttggaa
     1201 gaaaccgggc ggaacaggtc caaccggcat accatcccct ggcggaatat tgcccagtac
     1261 tggggaaggt gcggctgcag cgctataatc gtgaaacgcc tttgcctcac tgctgtgttc
     1321 gcaccgggaa gcggccgcac ttcccccgga gcctccgctt ccgcccttgt acagctcgtc
     1381 catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt gatcgcgctt
     1441 ctcgttgggg tctttgctca gcttggactg ggtgctcagg tagtggttgt cgggcagcag
     1501 cacggggccg tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct gcacgctgcc
     1561 gtcctcgatg ttgtggcgga tcttgaagtt caccttgatg ccgttcttct gcttgtcggc
     1621 catgatatag acgttgtggc tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc
     1681 gtcctccttg aagtcgatgc ccttcagctc gatgcggttc accagggtgt cgccctcgaa
     1741 cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg
     1801 gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta
     1861 gcggctgaag cactgcacgc cgtaggtcag ggtggtcacg agggtgggcc agggcacggg
     1921 cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg catcgccctc
     1981 gccctcgccg gacacgctga acttgtggcc gtttacgtcg ccgtccagct cgaccaggat
     2041 gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc atggtggcga ccggtggatc
     2101 ccgggcccgc ggtaccgtcg actgcagaat tcggttcact aaacgagccc ggttcactaa
     2161 acgagctctg cttatataga cctcccaccg tacacgccta ccgcccattt gcgtcaatgg
     2221 ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa acaaactccc
     2281 attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct atccacgccc
     2341 attgatgtac tgccaaaacc gcatcaccat ggtaatagcg atgactaata cgtagatgta
     2401 ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt
     2461 accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa
     2521 gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt
     2581 tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc
     2641 aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc tatgaactaa
     2701 tgaccccgta attgattact attaataact agcacgcgtt cgaagcttga gctcggtccc
     2761 ctcggggttg ggaggtgggt ctgaaacgat aatggtgaat atccctgcct aactctattc
     2821 actatagaaa gtacagcaaa aactattctt aaacctacca agcctcctac tatcattatg
     2881 aataatttta tataccacag ccaatttgtt atgttaaacc aattccacaa acttgcccat
     2941 ttatctaatt ccaataattc ttgttcattc ttttcttgct ggttttgcga ttcttcaatt
     3001 aaggagtgta ttaagcttgt gtaattgtta atttctctgt cccactccat ccaggtcgtg
     3061 tgattccaaa tctgttccag agatttatta ctccaactag cattccaagg cacagcagtg
     3121 gtgcaaatga gttttccaga gcaaccccaa atccccagga gctgttgatc ctttaggtat
     3181 ctttccacag ccaggattct tgcctggagc tgcttgatgc cccagactgt gagttgcaac
     3241 agatgctgtt gcgcctcaat agccctcagc aaattgttct gctgctgcac tataccagac
     3301 aataattgtc tggcctgtac cgtcagcgtc attgacgctg cgcccatagt gcttcctgct
     3361 gctcccaaga acccaaggaa caaagctcct attcccactg ctcttttttc tctctgcacc
     3421 actcttctct ttgccttggt gggtgctact cctaatggtt caatttttac tactttatat
     3481 ttatataatt cacttctcca attgtccctc atatctcctc cttatggtgt tttactaaac
     3541 ttttccatgt tctaatcctc atcctgtcta cttgccacac aatcatcacc tgccatctgt
     3601 tttccataat ccctaatgat ctttgctttt cttcttggca ctacttttat gtcactatta
     3661 tcttgtatta ctactgcccc ttcacctttc cagaggagct ttgctggtcc tttccaaagt
     3721 ggatttctgc tgtccctgta ataaacccga aaattttgaa tttttgtaat ttgtttttgt
     3781 aattctttag tttgtatgtc tgttgctatt atgtctacta ttctttcccc tgcactgtac
     3841 cccccaatcc ccccttttct tttaaaattg tggatgaata ctgccatttg tactgctgtc
     3901 ttaagatgtt cagcctgatc tcttacctgt cctataattt tctttaattc tttattcata
     3961 gattctacta ctccttgact ttggggattg tagggaattt ccaggtctga agatcctgct
     4021 gcttgctgtg cttttttctt acttttgttt tgctcttcct ctatcttgtc taaagcttcc
     4081 ttggtgtctt ttatctctat cctttgatgc acacaataga gggttgctac tgtattatat
     4141 aatgatctaa gttcttctga tcctgtctga agggatggtt gtagctgtcc cagtatttgt
     4201 ctacagcctt ctgatgtttc taacaggcca gtattaactg cgaatcgttc tagctccctg
     4261 cttgcccata ctatatgttt taatttatat tttttctttc cccctggcct taaccgaatt
     4321 ttttcccatc gcgatctaat tctcccccgc ttaatactga cgctctcgca cccatctctc
     4381 tccttctagc ctccgctagt caaaattttt ggcgtactca ccagtcgccg cccctcgcct
     4441 cttgccgtgc gcgcttcagc aagccgagtc ctgcgtcgag agagctcctc tggtttccct
     4501 ttcgctttca agtccctgtt cgggcgccac tgctagagat tttccacact gactaaaagg
     4561 gtctgaggga tctctagtta ccagagtcac acaacagacg ggcacacact acttgaagca
     4621 ctcaaggcaa gctttattga ggcttaagca gtgggttccc tagttagcca gagagctccc
     4681 aggctcagat ctggtctaac cagagagacc cagtacaggc aaaaagcagc tgcttatatg
     4741 caggatctga gggctcgcca ctccccagtc ccgcccaggc cacgcctccc atggtaatag
     4801 cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg
     4861 cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat
     4921 gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc
     4981 aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc
     5041 gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcggaact
     5101 ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa ctagtcaata
     5161 atcaatgtcg acccaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5221 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5281 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5341 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5401 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5461 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5521 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     5581 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5641 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5701 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5761 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5821 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     5881 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     5941 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     6001 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     6061 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     6121 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     6181 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6241 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6301 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6361 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6421 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6481 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6541 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6601 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6661 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6721 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6781 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6841 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6901 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6961 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     7021 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     7081 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     7141 tggccgattc attaatgcag cggatccaga catgataaga tacattgatg agtttggaca
     7201 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     7261 tttatttgta accattataa gctgcaataa acaagttagc tttttgcaaa agcctaggcc
     7321 tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg cctcggcctc
     7381 tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt
     7441 taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcgg
     7501 atccgctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     7561 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     7621 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     7681 gccaagcttg ggctgcaggt cgagggatct ccataagaga agagggacag ctatgactgg
     7741 gagtagtcag gagaggagga aaaatctggc tagtaaaaca tgtaaggaaa attttaggga
     7801 tgttaaagaa aaaaataaca caaaacaaaa tat
//
