LOCUS       sequence-488201-        7305 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_181, derived from human fusion protein SPAG9_SKAP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7305)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 7305)
  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_181
FEATURES             Location/Qualifiers
     source          1..7305
                     /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(838..1548,1549..1551,1552..1554))
                     /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(880..901)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1229..1248
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1488..1509
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1551..1560
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1636..1839)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1665..1685)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1840..2143
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(2400..2444)
                     /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    2629..2862
                     /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    3222..3339
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    3831..3956
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4003..4260
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        4262..4641
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4672..4776
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4777..4845,4846..5637)
                     /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(4995..5014)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5808..6396
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6297..6316
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6550..6567
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6635..6769
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6659..6678)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6713..6732
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6775..6970)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6789..6924
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    7029..7050
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7065..7070,7071..7088,7089..7095)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7103..7119
                     /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     7108..7130
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7127..7143
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7127..7143
                     /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 cttgcgacgt gtgccagact catcttcttc
      601 aagatcgtgc tcctcgtcag gtaagacttc atagatgtct tcctcctcct tctcttctgt
      661 tggctcttta atcccaactg aacccggcaa gatagtggga cgacattgac ttccgcacga
      721 gggagagtcg aacccatcga tgtcgtcata cgtctcctct ttctcctcct cctcctcgtc
      781 ctcttcataa gggatccggg aagcggccgc acttcccccg gagcctccgc ttccgccctt
      841 gtacagctcg tccatgccga gagtgatccc ggcggcggtc acgaactcca gcaggaccat
      901 gtgatcgcgc ttctcgttgg ggtctttgct cagcttggac tgggtgctca ggtagtggtt
      961 gtcgggcagc agcacggggc cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag
     1021 ctgcacgctg ccgtcctcga tgttgtggcg gatcttgaag ttcaccttga tgccgttctt
     1081 ctgcttgtcg gccatgatat agacgttgtg gctgttgtag ttgtactcca gcttgtgccc
     1141 caggatgttg ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt
     1201 gtcgccctcg aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat
     1261 ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg aagaagtcgt gctgcttcat
     1321 gtggtcgggg tagcggctga agcactgcac gccgtaggtc agggtggtca cgagggtggg
     1381 ccagggcacg ggcagcttgc cggtggtgca gatgaacttc agggtcagct tgccgtaggt
     1441 ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg ccgtttacgt cgccgtccag
     1501 ctcgaccagg atgggcacca ccccggtgaa cagctcctcg cccttgctca ccatggtggc
     1561 gaccggtgga tcccgggccc gcggtaccgt cgactgcaga attcggttca ctaaacgagc
     1621 ccggttcact aaacgagctc tgcttatata gacctcccac cgtacacgcc taccgcccat
     1681 ttgcgtcaat ggggcggagt tgttacgaca ttttggaaag tcccgttgat tttggtgcca
     1741 aaacaaactc ccattgacgt caatggggtg gagacttgga aatccccgtg agtcaaaccg
     1801 ctatccacgc ccattgatgt actgccaaaa ccgcatcacc atggtaatag cgatgactaa
     1861 tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca
     1921 ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg
     1981 atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt
     2041 ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt
     2101 gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg
     2161 gctatgaact aatgaccccg taattgatta ctattaataa ctagcacgcg ttcgaagctt
     2221 gagctcggtc ccctcggggt tgggaggtgg gtctgaaacg ataatggtga atatccctgc
     2281 ctaactctat tcactataga aagtacagca aaaactattc ttaaacctac caagcctcct
     2341 actatcatta tgaataattt tatataccac agccaatttg ttatgttaaa ccaattccac
     2401 aaacttgccc atttatctaa ttccaataat tcttgttcat tcttttcttg ctggttttgc
     2461 gattcttcaa ttaaggagtg tattaagctt gtgtaattgt taatttctct gtcccactcc
     2521 atccaggtcg tgtgattcca aatctgttcc agagatttat tactccaact agcattccaa
     2581 ggcacagcag tggtgcaaat gagttttcca gagcaacccc aaatccccag gagctgttga
     2641 tcctttaggt atctttccac agccaggatt cttgcctgga gctgcttgat gccccagact
     2701 gtgagttgca acagatgctg ttgcgcctca atagccctca gcaaattgtt ctgctgctgc
     2761 actataccag acaataattg tctggcctgt accgtcagcg tcattgacgc tgcgcccata
     2821 gtgcttcctg ctgctcccaa gaacccaagg aacaaagctc ctattcccac tgctcttttt
     2881 tctctctgca ccactcttct ctttgccttg gtgggtgcta ctcctaatgg ttcaattttt
     2941 actactttat atttatataa ttcacttctc caattgtccc tcatatctcc tccttatggt
     3001 gttttactaa acttttccat gttctaatcc tcatcctgtc tacttgccac acaatcatca
     3061 cctgccatct gttttccata atccctaatg atctttgctt ttcttcttgg cactactttt
     3121 atgtcactat tatcttgtat tactactgcc ccttcacctt tccagaggag ctttgctggt
     3181 cctttccaaa gtggatttct gctgtccctg taataaaccc gaaaattttg aatttttgta
     3241 atttgttttt gtaattcttt agtttgtatg tctgttgcta ttatgtctac tattctttcc
     3301 cctgcactgt accccccaat cccccctttt cttttaaaat tgtggatgaa tactgccatt
     3361 tgtactgctg tcttaagatg ttcagcctga tctcttacct gtcctataat tttctttaat
     3421 tctttattca tagattctac tactccttga ctttggggat tgtagggaat ttccaggtct
     3481 gaagatcctg ctgcttgctg tgcttttttc ttacttttgt tttgctcttc ctctatcttg
     3541 tctaaagctt ccttggtgtc ttttatctct atcctttgat gcacacaata gagggttgct
     3601 actgtattat ataatgatct aagttcttct gatcctgtct gaagggatgg ttgtagctgt
     3661 cccagtattt gtctacagcc ttctgatgtt tctaacaggc cagtattaac tgcgaatcgt
     3721 tctagctccc tgcttgccca tactatatgt tttaatttat attttttctt tccccctggc
     3781 cttaaccgaa ttttttccca tcgcgatcta attctccccc gcttaatact gacgctctcg
     3841 cacccatctc tctccttcta gcctccgcta gtcaaaattt ttggcgtact caccagtcgc
     3901 cgcccctcgc ctcttgccgt gcgcgcttca gcaagccgag tcctgcgtcg agagagctcc
     3961 tctggtttcc ctttcgcttt caagtccctg ttcgggcgcc actgctagag attttccaca
     4021 ctgactaaaa gggtctgagg gatctctagt taccagagtc acacaacaga cgggcacaca
     4081 ctacttgaag cactcaaggc aagctttatt gaggcttaag cagtgggttc cctagttagc
     4141 cagagagctc ccaggctcag atctggtcta accagagaga cccagtacag gcaaaaagca
     4201 gctgcttata tgcaggatct gagggctcgc cactccccag tcccgcccag gccacgcctc
     4261 ccatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg
     4321 tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg
     4381 tacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca
     4441 cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg
     4501 acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg
     4561 taacgcggaa ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactattaat
     4621 aactagtcaa taatcaatgt cgacccaggt ggcacttttc ggggaaatgt gcgcggaacc
     4681 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4741 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     4801 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     4861 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     4921 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     4981 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     5041 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     5101 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5161 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     5221 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5281 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     5341 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     5401 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     5461 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     5521 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     5581 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     5641 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5701 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5761 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5821 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5881 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5941 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6001 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6061 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6121 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6181 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6241 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6301 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6361 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6421 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     6481 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     6541 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     6601 ctccccgcgc gttggccgat tcattaatgc agcggatcca gacatgataa gatacattga
     6661 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     6721 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta gctttttgca
     6781 aaagcctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc
     6841 ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga
     6901 actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta
     6961 attgagatgc ggatccgctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     7021 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     7081 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     7141 gaccatgatt acgccaagct tgggctgcag gtcgagggat ctccataaga gaagagggac
     7201 agctatgact gggagtagtc aggagaggag gaaaaatctg gctagtaaaa catgtaagga
     7261 aaattttagg gatgttaaag aaaaaaataa cacaaaacaa aatat
//
