LOCUS       sequence-488167-        8301 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Express mEGFP-tagged intrinsically disordered protein (IDR), 
            FOidr_146, derived from human fusion protein PAX5_PAX5; ZCCHC7_PAX5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8301)
  TITLE     Kriwacki lab plasmids 2025
REFERENCE   2  (bases 1 to 8301)
  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_146
FEATURES             Location/Qualifiers
     source          1..8301
                     /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(1834..2544,2545..2547,2548..2550))
                     /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(1876..1897)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2225..2244
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2484..2505
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2547..2556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2632..2835)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2661..2681)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2836..3139
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(3396..3440)
                     /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    3625..3858
                     /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    4218..4335
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    4827..4952
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4999..5256
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     enhancer        5258..5637
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5668..5772
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5773..5841,5842..6633)
                     /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(5991..6010)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6804..7392
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7293..7312
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7546..7563
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7631..7765
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7655..7674)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7709..7728
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(7771..7966)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7785..7920
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    8025..8046
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8061..8066,8067..8084,8085..8091)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8099..8115
                     /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     8104..8126
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8123..8139
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8123..8139
                     /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 ataggacgaa tactggggat ggctataagg
      601 actcccggag aactccgatc ctggcaccat tcctgtaagg gtcggcgccg aatagctacc
      661 ctggcctgct ggtggaacgt gggggggata ccccggcagt gttgtcgatg ctaagtcgcg
      721 ccccgtcacg atagggtatg attgcgggcc gggcacgcta ctgccgatat cggcaggcgt
      781 tggactagcc agattagcct tcatgtcatc caggccgccg gcaagcgatg ccatcgcaga
      841 gtattcagtt gtctgctccg gcttaatggg ctcagtcgtt gtgaagatgt cagaataatg
      901 ttggcgttcg aatacgcggt ccagaacttc taattgctgt tgggtaaata agtccccgcg
      961 catctgttta cgcagaaaat cacgaccagg caaggaatga ccattcggga cgggtgattc
     1021 ttgaatcccc tcgtcacgct tacgcttgtt ggtatctgcg gaggggctgg tgattcctaa
     1081 aattccagaa atgctatacg aagatccagc actgtctgta ctcaccgacg agacttgagt
     1141 aacggagccg gtacttacaa tactatgcga gctcgctgga acgggttgat tcggcggctg
     1201 ctgcaccttc gtgcggataa tacgattgat cgacgatact gatggaacag tatcgttatc
     1261 acagacacgt tcggccagca agcgatcacg aatttcccag gcaaacatgg tcggattctg
     1321 gcgcttatac tctgcaattt tttcaacaac ctttggagta gctactttcg gttttgagcc
     1381 cccaatgacc cctggcttaa tgcttccggt ctcataatag cggcccaaga tctttgatac
     1441 gcaaccatgc gaaacacgca gctggcgact aatgtcgcat gggcgaacac cctggtgcgc
     1501 cagttcgacg atgcgttggc gtaccacatc cggcaaggga cgcccattaa caaatactcc
     1561 ccccaactgg ttcacgccgc cgtgcccttc gtcgcgcttg cgcttattcg tgtcggctga
     1621 agggcttgtg atccctaaga ttcctgagat actatacgat gaacctgccg aatcagtaga
     1681 aacagaggat acctgtgtta cagaaccggt ggacacaatc gagtggctcg atgccggtac
     1741 gggctggttt ggaggttgtt gaacttttgt acgaatgata cgattaatcg accgggaagc
     1801 ggccgcactt cccccggagc ctccgcttcc gcccttgtac agctcgtcca tgccgagagt
     1861 gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct cgttggggtc
     1921 tttgctcagc ttggactggg tgctcaggta gtggttgtcg ggcagcagca cggggccgtc
     1981 gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt
     2041 gtggcggatc ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca tgatatagac
     2101 gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt cctccttgaa
     2161 gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc
     2221 gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga cgtagccttc
     2281 gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca
     2341 ctgcacgccg taggtcaggg tggtcacgag ggtgggccag ggcacgggca gcttgccggt
     2401 ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga
     2461 cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc
     2521 ggtgaacagc tcctcgccct tgctcaccat ggtggcgacc ggtggatccc gggcccgcgg
     2581 taccgtcgac tgcagaattc ggttcactaa acgagcccgg ttcactaaac gagctctgct
     2641 tatatagacc tcccaccgta cacgcctacc gcccatttgc gtcaatgggg cggagttgtt
     2701 acgacatttt ggaaagtccc gttgattttg gtgccaaaac aaactcccat tgacgtcaat
     2761 ggggtggaga cttggaaatc cccgtgagtc aaaccgctat ccacgcccat tgatgtactg
     2821 ccaaaaccgc atcaccatgg taatagcgat gactaatacg tagatgtact gccaagtagg
     2881 aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac
     2941 gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt gggcagttta
     3001 ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac
     3061 atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt
     3121 taccgtaagt tatgtaacgc ggaactccat atatgggcta tgaactaatg accccgtaat
     3181 tgattactat taataactag cacgcgttcg aagcttgagc tcggtcccct cggggttggg
     3241 aggtgggtct gaaacgataa tggtgaatat ccctgcctaa ctctattcac tatagaaagt
     3301 acagcaaaaa ctattcttaa acctaccaag cctcctacta tcattatgaa taattttata
     3361 taccacagcc aatttgttat gttaaaccaa ttccacaaac ttgcccattt atctaattcc
     3421 aataattctt gttcattctt ttcttgctgg ttttgcgatt cttcaattaa ggagtgtatt
     3481 aagcttgtgt aattgttaat ttctctgtcc cactccatcc aggtcgtgtg attccaaatc
     3541 tgttccagag atttattact ccaactagca ttccaaggca cagcagtggt gcaaatgagt
     3601 tttccagagc aaccccaaat ccccaggagc tgttgatcct ttaggtatct ttccacagcc
     3661 aggattcttg cctggagctg cttgatgccc cagactgtga gttgcaacag atgctgttgc
     3721 gcctcaatag ccctcagcaa attgttctgc tgctgcacta taccagacaa taattgtctg
     3781 gcctgtaccg tcagcgtcat tgacgctgcg cccatagtgc ttcctgctgc tcccaagaac
     3841 ccaaggaaca aagctcctat tcccactgct cttttttctc tctgcaccac tcttctcttt
     3901 gccttggtgg gtgctactcc taatggttca atttttacta ctttatattt atataattca
     3961 cttctccaat tgtccctcat atctcctcct tatggtgttt tactaaactt ttccatgttc
     4021 taatcctcat cctgtctact tgccacacaa tcatcacctg ccatctgttt tccataatcc
     4081 ctaatgatct ttgcttttct tcttggcact acttttatgt cactattatc ttgtattact
     4141 actgcccctt cacctttcca gaggagcttt gctggtcctt tccaaagtgg atttctgctg
     4201 tccctgtaat aaacccgaaa attttgaatt tttgtaattt gtttttgtaa ttctttagtt
     4261 tgtatgtctg ttgctattat gtctactatt ctttcccctg cactgtaccc cccaatcccc
     4321 ccttttcttt taaaattgtg gatgaatact gccatttgta ctgctgtctt aagatgttca
     4381 gcctgatctc ttacctgtcc tataattttc tttaattctt tattcataga ttctactact
     4441 ccttgacttt ggggattgta gggaatttcc aggtctgaag atcctgctgc ttgctgtgct
     4501 tttttcttac ttttgttttg ctcttcctct atcttgtcta aagcttcctt ggtgtctttt
     4561 atctctatcc tttgatgcac acaatagagg gttgctactg tattatataa tgatctaagt
     4621 tcttctgatc ctgtctgaag ggatggttgt agctgtccca gtatttgtct acagccttct
     4681 gatgtttcta acaggccagt attaactgcg aatcgttcta gctccctgct tgcccatact
     4741 atatgtttta atttatattt tttctttccc cctggcctta accgaatttt ttcccatcgc
     4801 gatctaattc tcccccgctt aatactgacg ctctcgcacc catctctctc cttctagcct
     4861 ccgctagtca aaatttttgg cgtactcacc agtcgccgcc cctcgcctct tgccgtgcgc
     4921 gcttcagcaa gccgagtcct gcgtcgagag agctcctctg gtttcccttt cgctttcaag
     4981 tccctgttcg ggcgccactg ctagagattt tccacactga ctaaaagggt ctgagggatc
     5041 tctagttacc agagtcacac aacagacggg cacacactac ttgaagcact caaggcaagc
     5101 tttattgagg cttaagcagt gggttcccta gttagccaga gagctcccag gctcagatct
     5161 ggtctaacca gagagaccca gtacaggcaa aaagcagctg cttatatgca ggatctgagg
     5221 gctcgccact ccccagtccc gcccaggcca cgcctcccat ggtaatagcg atgactaata
     5281 cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg
     5341 cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat
     5401 gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc
     5461 ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg
     5521 gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc
     5581 tatgaactaa tgaccccgta attgattact attaataact agtcaataat caatgtcgac
     5641 ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5701 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5761 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5821 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5881 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5941 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     6001 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     6061 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     6121 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     6181 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     6241 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     6301 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     6361 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     6421 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     6481 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6541 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6601 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6661 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6721 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6781 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6841 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6901 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     6961 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     7021 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     7081 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     7141 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     7201 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     7261 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     7321 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7381 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     7441 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     7501 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     7561 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     7621 taatgcagcg gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     7681 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     7741 cattataagc tgcaataaac aagttagctt tttgcaaaag cctaggcctc caaaaaagcc
     7801 tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa
     7861 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat
     7921 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcggat ccgctggcac
     7981 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     8041 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     8101 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcttggg
     8161 ctgcaggtcg agggatctcc ataagagaag agggacagct atgactggga gtagtcagga
     8221 gaggaggaaa aatctggcta gtaaaacatg taaggaaaat tttagggatg ttaaagaaaa
     8281 aaataacaca aaacaaaata t
//
