LOCUS       sequence-487932-        8421 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_014, derived from human fusion protein ATF7IP_JAK2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8421)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8421)
  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_014
FEATURES             Location/Qualifiers
     source          1..8421
                     /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(1954..2664,2665..2667,2668..2670))
                     /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(1996..2017)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2345..2364
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2604..2625
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2667..2676
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2752..2955)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2781..2801)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2956..3259
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3516..3560)
                     /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    3745..3978
                     /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    4338..4455
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4947..5072
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5119..5376
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5378..5757
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5788..5892
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5893..5961,5962..6753)
                     /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(6111..6130)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6924..7512
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7413..7432
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7666..7683
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7751..7885
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7775..7794)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7829..7848
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7891..8086)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7905..8040
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    8145..8166
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8181..8186,8187..8204,8205..8211)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8219..8235
                     /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     8224..8246
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8243..8259
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8243..8259
                     /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 tctctcctga agaatatttt ggggattgac
      601 ctgccgaaca gcaggagcct gcataacagt tcctcgcaag ggagcctgag ctggtggtgc
      661 aggaagagtt gtataaacca cctggtggtt tgcaggagct cttggtacag ggagtcttgt
      721 ggtcacctga gttactggac gatgtgttac attcacggta gttggagctg taggtagtaa
      781 gtgtatggtg gtctgagatg gtccacttgt tggcacccca gatggttgaa gaggcggtgc
      841 tggttgtatg ggttgcaatg gtcgtgacac aggctgagaa gaactcatgg ttgatacagg
      901 agtgtgattt agttttttgg ggtcttgtga agctccactc tcttcatcat ccattgtgag
      961 atcaatgaca ccacttgaat cactgccagt ggcttttcca cactgtgatg tgctatcagc
     1021 tgctttctta ctgacagaag catctattgt tttgtttgtc tggttttcaa ttcttggggt
     1081 ggtattgctg ttctgttctg cagcagacga agtactaaat gcagaaattg gaattttcat
     1141 ctgtatagga cctcgattag ttgatggact atagagtcct gatgggccca ctgtgggtaa
     1201 agaagtcctt gtggcttgta caatagagtg tgctgttgga acagcctgca cagcaagtgt
     1261 tgttcccaat ggtgcagcac tggcagtagg gttcctttga atactaggac tgggcacaga
     1321 aggaacgttg tttggtttag tgggatttgg caaggatggc aaggatactg gatttttggt
     1381 aagaccactc actgtaggtg ggctttgcac agaaatgaat tcaacattgc cagatggttg
     1441 attagtcacc aaagtccctg gatggctctg tagaagctga ggctgggagg atactgcaac
     1501 aggtacatgc aaaatcactg gcaaaggctg taaagagatt gtaggctggg gatttccact
     1561 aggcacctga gtagtagcaa ctacagtagc tgtattgggt gcaggaagaa ctgacgttgc
     1621 tgtgagggaa cccgaagtca ctggagtctg caatttaggt tgactactga caactgctgg
     1681 aggagtgaca agattggttg aagatactgt attcacagga gtttgaaagg atggtggtgc
     1741 actctcgctt acatttcttt tggactccag catctgtctc actgtgcctg catttctgta
     1801 agacatgtta ttattgctgt tgacatcatt tacagttttt cctggtgata ctggtgggtt
     1861 gggtggatgt tcatgtcttt tcttaagatc ttctttggct gcttcaaagc gccgggaagc
     1921 ggccgcactt cccccggagc ctccgcttcc gcccttgtac agctcgtcca tgccgagagt
     1981 gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct cgttggggtc
     2041 tttgctcagc ttggactggg tgctcaggta gtggttgtcg ggcagcagca cggggccgtc
     2101 gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt
     2161 gtggcggatc ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca tgatatagac
     2221 gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt cctccttgaa
     2281 gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc
     2341 gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga cgtagccttc
     2401 gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca
     2461 ctgcacgccg taggtcaggg tggtcacgag ggtgggccag ggcacgggca gcttgccggt
     2521 ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga
     2581 cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc
     2641 ggtgaacagc tcctcgccct tgctcaccat ggtggcgacc ggtggatccc gggcccgcgg
     2701 taccgtcgac tgcagaattc ggttcactaa acgagcccgg ttcactaaac gagctctgct
     2761 tatatagacc tcccaccgta cacgcctacc gcccatttgc gtcaatgggg cggagttgtt
     2821 acgacatttt ggaaagtccc gttgattttg gtgccaaaac aaactcccat tgacgtcaat
     2881 ggggtggaga cttggaaatc cccgtgagtc aaaccgctat ccacgcccat tgatgtactg
     2941 ccaaaaccgc atcaccatgg taatagcgat gactaatacg tagatgtact gccaagtagg
     3001 aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac
     3061 gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt gggcagttta
     3121 ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac
     3181 atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt
     3241 taccgtaagt tatgtaacgc ggaactccat atatgggcta tgaactaatg accccgtaat
     3301 tgattactat taataactag cacgcgttcg aagcttgagc tcggtcccct cggggttggg
     3361 aggtgggtct gaaacgataa tggtgaatat ccctgcctaa ctctattcac tatagaaagt
     3421 acagcaaaaa ctattcttaa acctaccaag cctcctacta tcattatgaa taattttata
     3481 taccacagcc aatttgttat gttaaaccaa ttccacaaac ttgcccattt atctaattcc
     3541 aataattctt gttcattctt ttcttgctgg ttttgcgatt cttcaattaa ggagtgtatt
     3601 aagcttgtgt aattgttaat ttctctgtcc cactccatcc aggtcgtgtg attccaaatc
     3661 tgttccagag atttattact ccaactagca ttccaaggca cagcagtggt gcaaatgagt
     3721 tttccagagc aaccccaaat ccccaggagc tgttgatcct ttaggtatct ttccacagcc
     3781 aggattcttg cctggagctg cttgatgccc cagactgtga gttgcaacag atgctgttgc
     3841 gcctcaatag ccctcagcaa attgttctgc tgctgcacta taccagacaa taattgtctg
     3901 gcctgtaccg tcagcgtcat tgacgctgcg cccatagtgc ttcctgctgc tcccaagaac
     3961 ccaaggaaca aagctcctat tcccactgct cttttttctc tctgcaccac tcttctcttt
     4021 gccttggtgg gtgctactcc taatggttca atttttacta ctttatattt atataattca
     4081 cttctccaat tgtccctcat atctcctcct tatggtgttt tactaaactt ttccatgttc
     4141 taatcctcat cctgtctact tgccacacaa tcatcacctg ccatctgttt tccataatcc
     4201 ctaatgatct ttgcttttct tcttggcact acttttatgt cactattatc ttgtattact
     4261 actgcccctt cacctttcca gaggagcttt gctggtcctt tccaaagtgg atttctgctg
     4321 tccctgtaat aaacccgaaa attttgaatt tttgtaattt gtttttgtaa ttctttagtt
     4381 tgtatgtctg ttgctattat gtctactatt ctttcccctg cactgtaccc cccaatcccc
     4441 ccttttcttt taaaattgtg gatgaatact gccatttgta ctgctgtctt aagatgttca
     4501 gcctgatctc ttacctgtcc tataattttc tttaattctt tattcataga ttctactact
     4561 ccttgacttt ggggattgta gggaatttcc aggtctgaag atcctgctgc ttgctgtgct
     4621 tttttcttac ttttgttttg ctcttcctct atcttgtcta aagcttcctt ggtgtctttt
     4681 atctctatcc tttgatgcac acaatagagg gttgctactg tattatataa tgatctaagt
     4741 tcttctgatc ctgtctgaag ggatggttgt agctgtccca gtatttgtct acagccttct
     4801 gatgtttcta acaggccagt attaactgcg aatcgttcta gctccctgct tgcccatact
     4861 atatgtttta atttatattt tttctttccc cctggcctta accgaatttt ttcccatcgc
     4921 gatctaattc tcccccgctt aatactgacg ctctcgcacc catctctctc cttctagcct
     4981 ccgctagtca aaatttttgg cgtactcacc agtcgccgcc cctcgcctct tgccgtgcgc
     5041 gcttcagcaa gccgagtcct gcgtcgagag agctcctctg gtttcccttt cgctttcaag
     5101 tccctgttcg ggcgccactg ctagagattt tccacactga ctaaaagggt ctgagggatc
     5161 tctagttacc agagtcacac aacagacggg cacacactac ttgaagcact caaggcaagc
     5221 tttattgagg cttaagcagt gggttcccta gttagccaga gagctcccag gctcagatct
     5281 ggtctaacca gagagaccca gtacaggcaa aaagcagctg cttatatgca ggatctgagg
     5341 gctcgccact ccccagtccc gcccaggcca cgcctcccat ggtaatagcg atgactaata
     5401 cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg
     5461 cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat
     5521 gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc
     5581 ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg
     5641 gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc
     5701 tatgaactaa tgaccccgta attgattact attaataact agtcaataat caatgtcgac
     5761 ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5821 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5881 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5941 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     6001 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     6061 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     6121 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     6181 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     6241 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     6301 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     6361 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     6421 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     6481 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     6541 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     6601 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6661 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6721 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6781 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6841 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6901 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6961 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     7021 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     7081 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     7141 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     7201 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     7261 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     7321 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     7381 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     7441 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7501 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     7561 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     7621 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     7681 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     7741 taatgcagcg gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     7801 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     7861 cattataagc tgcaataaac aagttagctt tttgcaaaag cctaggcctc caaaaaagcc
     7921 tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa
     7981 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat
     8041 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcggat ccgctggcac
     8101 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     8161 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     8221 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcttggg
     8281 ctgcaggtcg agggatctcc ataagagaag agggacagct atgactggga gtagtcagga
     8341 gaggaggaaa aatctggcta gtaaaacatg taaggaaaat tttagggatg ttaaagaaaa
     8401 aaataacaca aaacaaaata t
//
